bromochloroacetic-acid and Psoriasis

Prolactin (PRL) is well recognised for its role(s) in mammary gland development and function. Moreover, its role in skin biology, including the potent regulation of human hair growth, is becoming clearer. Less widely appreciated, however, is the potential role of PRL in the pathobiology of psoriasis. While the relationship between PRL and psoriasis remains enigmatic, several recent publications on the PRL-psoriasis connection have demonstrated a reawakening of interest in this conundrum. We take the occasion of these reports to underscore the importance of dissecting the role(s) of PRL in the aetiopathology of psoriasis, not least since this may help to identify novel hormonal treatment strategies in its management.

Topics: Animals; Cell Growth Processes; Gene Expression Regulation; Hair; Humans; Immunomodulation; Keratins; Prolactin; Psoriasis; Skin; Stress, Psychological

Psoriasis is strongly associated with streptococcal throat infection, and patients have increased occurrence of such infections. Psoriatic lesional T cells are oligoclonal, and T cells recognizing determinants common to streptococcal M-protein and keratin have been detected in patients' blood. We propose that CD8(+) T cells in psoriatic epidermis respond mainly to such determinants, whereas CD4(+) T cells in the dermis preferentially recognize determinants on the streptococcal peptidoglycan that might itself act as an adjuvant. The streptococcal association might reflect the concurrence of superantigen production promoting skin-homing of tonsil T cells, M-protein mimicking keratin determinants, and adjuvant effects of the peptidoglycan. Accordingly, improvement of psoriasis after tonsillectomy should be associated with fewer T cells that recognize keratin and streptococcal determinants.

Topics: Antigens, Bacterial; Autoimmune Diseases; Bacterial Outer Membrane Proteins; Carrier Proteins; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Dermis; Epidermis; Humans; Keratins; Molecular Mimicry; Palatine Tonsil; Peptidoglycan; Psoriasis; Streptococcal Infections; Streptococcus; Superantigens; Tonsillitis

The use of DNA microarray technology in biomedical research has dramatically increased during the past years. In the present report, we provide an overview on the basic DNA microarray technology and biostatistical methods for gene expression analysis. A focus is then put on its applications in dermatological research. In recent years, a series of gene expression studies have been performed for various dermatological diseases, such as malignant melanoma, psoriasis and lupus erythematosus. These analyses have identified interesting target genes as well as putative disease susceptibility loci. However, further functional studies will be needed for a more complete understanding of the pathogenesis of these diseases. This may be performed by means of the recently developed RNA interference technology. Besides its role in large-scale gene expression studies, DNA microarray technology has proved to be a valuable tool for genomic screens of genetic alterations, e.g. single nucleotide polymorphisms. These play a role in tumour development and progression, and also function as genetic markers for disease susceptibility. Taken together, DNA microarray technology opens enormous perspectives for dermatologists. It may help us understand the complex pathogenesis of a wide variety of dermatologic diseases and identify their genetic background.

Topics: Cell Differentiation; Gene Expression Profiling; Humans; Hybridization, Genetic; Keratinocytes; Keratins; Melanoma; Oligonucleotide Array Sequence Analysis; Psoriasis; RNA, Messenger; Skin Neoplasms; Ultraviolet Rays

The evidence that T lymphocytes play a key role in the pathogenesis of psoriasis is now compelling. Eruption of psoriatic skin lesions coincides with epidermal infiltration and activation of T cells, and spontaneous or treatment-induced resolution of the lesions is preceded by the reduction or disappearance of epidermal T cells. An upregulation has also been demonstrated for various molecules associated with T-cell mediated inflammation, and treatments selectively directed against T cells have proved very effective. Infections with group A beta-haemolytic streptococci have been associated with onset of acute psoriasis and exacerbation of chronic psoriasis. Such infections are also frequently accompanied by erythematous skin rashes. Also, recent reports indicate that streptococcal superantigens can induce expression of cutaneous lymphocyte antigens (CLA), believed to play a major role in enabling T cells to migrate to the skin. Furthermore, T-cell lines isolated from psoriatic lesions may show strong reactivity to streptococcal antigens. We have postulated that psoriasis is an autoimmune disease mediated by T cells reacting to epitopes that are common to streptococcal M-proteins and keratins. To investigate this possibility, circulating T cells from 12 patients with active psoriasis, paired controls, and six patients with atopic dermatitis were challenged in vitro with five synthetic 20aa (amino acid) M-peptides: production of IFN-gamma and IL-4 was analysed by ELISPOT and RT-PCR techniques. Four of these peptides shared five to six aa sequences with several type I keratins and one did not. In 10 of the 12 psoriasis patients, measurable IFN-gamma production could be induced by one or more of the four peptides that share sequences with keratins. A borderline response was observed in only four of the 18 controls: the dermatitis patients were all negative. The only peptide that shared 6aa with keratins was the one that induced a response in the psoriatic patients most frequently, and four of them showed the strongest response to this peptide while none of the controls reacted to it. In all instances negligible responses were observed to the control peptide that did not share sequences with keratins. Except for PHA-stimulated controls, IL-4 responses could not be detected by either ELISPOT or RT-PCR and there was generally good agreement between the two techniques. A marked reduction in the M-peptide-induced IFN-gamma responses was observed in the psoriasis pa

Topics: Animals; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Carrier Proteins; Cross Reactions; Humans; Keratins; Psoriasis; Receptors, Antigen, T-Cell, alpha-beta; Streptococcus; Superantigens; T-Lymphocytes

Vitamin D3 analogs have been found to be effective in treating psoriasis.. We attempted to identify the targets and actions of vitamin D3 in the skin and to explore the availability of synthetic vitamin D3 analogs with selective actions on particular cell types or cell functions.. A review of the literature focused on the cellular targets of vitamin D3 in the skin and within the immune system. Furthermore, the use of novel vitamin D3 analogs in skin diseases other than psoriasis was reviewed.. The vitamin D receptor has been detected in most skin cells, which means that keratinization, hair growth, melanogenesis, fibrogenesis, angiogenesis, and immune-mediated processes are potential targets for vitamin D3. Vitamin D3 analogs have been synthesized with a higher therapeutic index or a higher degree of selectivity than the natural form of vitamin D3.. Vitamin D3 analogs with wide-ranging clinical applications may become available for dermatology.

Topics: Adjuvants, Immunologic; Biological Availability; Cholecalciferol; Connective Tissue; Dermatologic Agents; Forecasting; Hair; Humans; Immunity, Cellular; Keratins; Melanins; Neovascularization, Physiologic; Psoriasis; Receptors, Calcitriol; Skin; Skin Diseases

Epithelial cell intermediate filaments, or cytokeratins, are excellent markers for cell differentiation. During embryogenesis, cytokeratins specific of a stage of differentiation step always become detectable before corresponding morphologic changes: for instance, cytokeratins 5 and 14 are found around the eight week, shortly before stratification of the epithelium occurs, and cytokeratins 1 and 10 are produced before morphologic evidence of keratinization becomes detectable. Among potential diagnostic applications, analysis of cytokeratin patterns of epidermal cells desquamated in the amniotic fluid may provide earlier and less invasive diagnosis than fetoscopic biopsies. Similarly, a review of cytokeratins expressed in a variety of epithelial diseases (involving the epidermis, digestive tract, respiratory tract, urogenital tract, or breast) demonstrated persistence of the original tissue pattern in some instances (this was the case for the majority of simple epithelia) but not in others (complex epithelia). This suggests that cytokeratins may prove valuable as markers for specific tumor stages or types and may provide earlier information than morphologic studies.

Topics: Breast Neoplasms; Digestive System Neoplasms; Epidermis; Female; Genital Neoplasms, Female; Genital Neoplasms, Male; Humans; Keratins; Male; Psoriasis; Skin Neoplasms

Topics: Biomarkers; Cell Differentiation; Epidermal Cells; Epithelial Cells; Keratinocytes; Keratins; Psoriasis

Topics: Arachidonic Acids; Calcium; Cell Differentiation; Cell Division; Epidermal Cells; Growth Substances; Humans; In Vitro Techniques; Insulin; Keratins; Nucleotides, Cyclic; Psoriasis; Receptors, Cell Surface; Somatostatin

Topics: Animals; Epidermis; Humans; Keratins; Psoriasis; Reference Values; Retinoids; Skin Neoplasms

Topics: Animals; Calcitriol; Carcinogens; Cells, Cultured; Epidermal Cells; Epidermis; Humans; Keratins; Mice; Psoriasis; Reference Values

Topics: Animals; Base Sequence; Cells, Cultured; DNA; Electrophoresis, Polyacrylamide Gel; Humans; Keratins; Mice; Protein Precursors; Psoriasis; Skin

The retinoids are synthetic derivatives of vitamin A. Isotretinoin (13-cis-retinoic acid) is now being widely used in the United States for severe acne and etretinate is available in Europe and other countries for psoriasis. These drugs are also effective for a number of other skin diseases. This is an attempt to review basic knowledge of retinoids with which the practicing dermatologist should be familiar, to review the current status of studies, and to speculate on the present and future roles of these drugs in dermatology.

Topics: Acne Vulgaris; Etretinate; Humans; Inflammation; Isotretinoin; Keratins; Psoriasis; Retinoids; Sebum; Skin Diseases; Skin Neoplasms; Sweat Gland Diseases; Tretinoin; Vitamin A

Topics: Adrenal Cortex Hormones; Cell Cycle; Cyclic AMP; Epidermis; Humans; Keratins; Keratosis; Protein Binding; Protein Precursors; Psoriasis

Topics: Acne Vulgaris; Adolescent; Child; Child, Preschool; Darier Disease; Etretinate; Female; Humans; Ichthyosis; Infant; Isomerism; Isotretinoin; Keratins; Keratoderma, Palmoplantar; Male; Pityriasis Rubra Pilaris; Psoriasis; Skin Diseases; Skin Diseases, Vesiculobullous; Tretinoin

Topics: Acrodermatitis; Amino Acid Metabolism, Inborn Errors; Basal Cell Nevus Syndrome; Epidermolysis Bullosa; Hair Diseases; Humans; Ichthyosis; Keratins; Keratosis; Neurofibromatosis 1; Psoriasis; Refsum Disease; Skin; Skin Diseases; Skin Neoplasms; Tuberous Sclerosis; Tyrosine; Warts; Xeroderma Pigmentosum

Topics: Adenylyl Cyclases; Arachidonic Acids; Calmodulin; Cell Division; Cyclic AMP; Cyclic GMP; Epidermis; Glycogen; Glycolysis; History, 20th Century; Homeostasis; Humans; Keratins; Lithium; Phosphorylation; Plasminogen Activators; Polyamines; Propranolol; Psoriasis; Receptors, Cell Surface; Skin

Topics: Fatty Acids; Fatty Acids, Essential; Humans; Hydrocarbons; Ichthyosis; Keratins; Lipid Metabolism; Lipids; Phospholipids; Psoriasis; Skin; Skin Diseases; Skin Physiological Phenomena; Sphingolipids; Sterols; Subcellular Fractions; Waxes

Topics: Animals; Cell Communication; Cell Differentiation; Cell Division; Cells, Cultured; Chick Embryo; Ectoderm; Epidermal Cells; Epidermis; Humans; Ichthyosis; Keratins; Mutation; Psoriasis; Skin Abnormalities

Topics: Adenylyl Cyclases; Cyclic AMP; Epidermis; Epinephrine; Growth Inhibitors; Homeostasis; Humans; Hyperplasia; Keratins; Mitosis; Phosphoric Diester Hydrolases; Psoriasis; Skin

Enzymic activities and cofactor levels in the epidermis are reviewed with special regard to psoriasis and the papulosquamous disorders lichen simplex and lichen planus. The metabolism of nicotinamide adenine dinucleotide phosphate and its dependent pathways seems to deviate in psoriasis from that in the contrasted dermatoses, in normal skin in health and in skin during the process of wound healing. The mitochondrial function also differs between psoriatic and normal skin. In some conditions in psoriasis this function cannot be seen to deviate from normality, however. The regulation and control of mitochondrial function in psoriasis might be another area in which future investigations may yield significant information on the pathophysiology in this skin disease.

Topics: Adenosine Triphosphate; Carbohydrate Metabolism; Citric Acid Cycle; Cytosol; Glycolysis; Humans; Keratins; Lichen Planus; Lipid Metabolism; Mitochondria; NAD; NADP; Oxidoreductases; Oxygen Consumption; Psoriasis; Skin

Numerous general metabolic systems are disturbed in association with psoriasis: the frequency of diabetes mellitus and of hyperuricaemia, lipid disturbances and a decrease in folates as a result of their excessive consumption by the skin. Cutaneous metabolism is also altered. Numerous compounds are formed in excess from glucose: amino acids, fatty acids and sterols, lactic acid--the formation of which persists in the corneal layer, ribose and ribulose--synthesised as a result of glucose-6-phosphate-dehydrogenase hyperactivity (role of the increased catabolism of dehydro-epi-androsterone) and uronic acids. The accumulation of glycogen is probably due to excessive synthesis and impaired breakdown. These abnormalities may exist to a lesser extent in healthy skin. In the corneal layer there are lipid vacuoles visible under the electron microscope. Lipogenesis is increased. The same may apply to lipolysis (blood NEFA are increased). Esterification of cholesterol is decreased. The utilisation of ATP by cell membranes is probably diminished (low ATP ase activity). The absence of formation of keratohyaline is due to persistence of the repression which normally prevents it in the mucus body. Renewal of collagen appears increased. The synthesis of DNA is increased in the lesions and neighbouring areas. It is possible that these various abnormalities are dependent upon modifications in the regulator systems of cyclic AMP and GMP, variations in which are however discussed.

Topics: Adenosine Triphosphatases; Blood Glucose; Collagen; Cyclic AMP; Cyclic GMP; DNA; Glycogen; Glycolysis; Humans; Keratins; Lipid Metabolism; Metabolic Diseases; Metals; Mitochondria; Oxygen Consumption; Pentosephosphates; Proteins; Psoriasis; Skin

Topics: Animals; Arachidonic Acids; Chemical Phenomena; Chemistry; Dermatitis; Humans; Keratins; Prostaglandins; Psoriasis; Skin; Ultraviolet Rays

Topics: Acne Vulgaris; Animals; Dermatologic Agents; Disease Models, Animal; Epithelial Cells; Epithelium; Female; Humans; Keratins; Mice; Mitosis; Psoriasis; Swine; Vagina

Topics: Autoradiography; Diet Therapy; DNA; Genetics, Medical; Humans; Keratins; Microscopy, Electron; Mitosis; Oxygen Consumption; Psoriasis; Skin; Skin Transplantation; Transplantation, Autologous; Zinc

Topics: Cholesterol; Eczema; Fatty Acids; Fatty Acids, Essential; Humans; Ichthyosis; Keratins; Lipoproteins; Psoriasis; Refsum Disease; Skin; Sterols

Topics: Chondroitin; Dermatitis, Exfoliative; Glycosaminoglycans; Hair; Homocystinuria; Humans; Keratins; Methionine; Psoriasis; Skin Diseases; Sulfates; Sulfur

In psoriatic skin, epidermal keratinocytes undergo deregulated inflammatory response that leads to prolonged expression of inflammatory mediators as well as abnormal keratins. Methotrexate (MTX) is an immunosuppressive agent used as a standard drug to treat severe psoriasis. The aim of the study is to find the pharmacological effect of MTX on abnormal keratin and deregulated inflammatory mediators.. Fifty-eight psoriasis vulgaris patients were recruited for this study. Skin biopsies of psoriatic patients were collected and analyzed for activation signal such as TNF-α and IFN-γ and deactivation signal such as TGF-β1. Also, protein and gene expression of normal keratins K14 and K10 and abnormal keratins K16 and K17 were analyzed in skin biopsies before (day 0) and after (at the end of 6 and 12 weeks) MTX treatment.. Results show a significant decrease in tissue TNF-α and IFN-γ and increase in TGF-β1 after MTX treatment when compared with before MTX treatment in psoriasis patients (p<0.001). Protein and gene expression of K14, K16 and K17 decreased after MTX treatment, whereas the expression of differentiation marker K10 increased after MTX treatment.. MTX resolves deregulated inflammatory markers and maintains normal keratin phenotype on hyperproliferating KC, thereby controlling acanthosis in psoriasis patients.

Topics: Adolescent; Adult; Aged; Cell Cycle; Down-Regulation; Female; Humans; Inflammation Mediators; Keratinocytes; Keratins; Male; Methotrexate; Middle Aged; Psoriasis; Young Adult

Exacerbation of chronic psoriasis can be associated with streptococcal throat infections, and T cells that respond to peptide sequences common to streptococcal M proteins and skin keratins have been detected in patients' blood. To our knowledge, we have conducted the first blinded, prospective study to assess the impact of tonsillectomy on psoriasis. Twenty-nine patients with chronic psoriasis and history of exacerbation after sore throat were randomly assigned to tonsillectomy (n = 15) or control (n = 14) groups and monitored for 2 y clinically and by enumeration of circulating skin homing T cells that respond to short homologous M protein or keratin peptides. Thirteen patients (86%) showed sustained improvement after tonsillectomy ranging from 30 to 90% reduction in disease severity. Furthermore, there was a close correlation between the degree of clinical improvement in individual patients and reduction in the frequency of peptide-reactive skin-homing T cells in their circulation. No corresponding clinical or immunologic changes were observed among the controls. These findings indicate that tonsillectomy may have a beneficial effect on chronic psoriasis because the palatine tonsils generate effector T cells that recognize keratin determinants in the skin.

Topics: Adolescent; Adult; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Carrier Proteins; Cell Movement; Child; Child, Preschool; Chronic Disease; Epitopes, T-Lymphocyte; Female; Humans; Keratins; Lymphopenia; Male; Middle Aged; Palatine Tonsil; Prospective Studies; Psoriasis; Skin; T-Lymphocyte Subsets; Tonsillectomy; Young Adult

Acitretin has been shown to be effective for psoriasis treatment. Its mechanism of action is not completely understood, and there are few studies focusing on histological and immunohistochemical differences before and after treatment of psoriasis with acitretin.. This is a prospective study of 17 patients with plaque psoriasis treated with acitretin for 4 months with biopsies taken before and after therapy. Histological features and immunohistochemical reactions to cytokeratin (CK) 10, CK16, CK19, Ki67 and CD1a were evaluated and compared.. There were nine men and eight women with median age of 47 years. Epidermal thickness, CK16 positivity, Ki67 and CD1a-positive cell index reduced after treatment (p < 0.01). Suprapapillary plate thickness stayed the same (p > 0.05) although the epidermal/suprapapillary thickness ratio was significantly higher before treatment (p < 0.01). CK10 positivity was lower and a thicker basal cell layer was seen in the epidermis before treatment (p < 0.01). CK19 was negative in all cases.. Acitretin therapy improved histological and immunohistochemical features typical of psoriasis. In psoriasis, suprapapillary plates are not thin, but the epidermal/suprapapillary thickness ratio is increased. Basal cell layer is expanded in psoriasis. Langerhans' cells were less frequent after treatment, and that finding has to be investigated further to determine its role in acitretin mechanism of action.

Topics: Acitretin; Adult; Biomarkers; Biopsy; Cell Count; Female; Humans; Keratinocytes; Keratins; Keratolytic Agents; Langerhans Cells; Male; Middle Aged; Prospective Studies; Psoriasis; Skin; Treatment Outcome

Calcitriol and calcipotriol are widely used in the topical treatment of psoriasis. However, studies comparing both treatment modalities are scarce. Especially, there are almost no studies comparing the effects on epidermal cell populations in a quantitative manner.. The aim of this study was to quantitatively compare the effects of topical calcitriol and topical calcipotriol on clinical scores and epidermal subpopulations.. From five patients with stable plaque psoriasis, skin biopsies were taken from two symmetrical regions on the trunk or extremities before and after treatment with either calcitriol or calcipotriol. Frozen sections were labelled immunofluorescently using direct immunofluorescence for beta-1 integrin and the Zenon labelling technique for keratin (K) 6, K10 and K15. The digital photographs of the stained sections were quantitatively analysed and the results of both treatments were compared.. The clinical SUM-score improved significantly for both the calcitriol- and the calcipotriol-treated lesions. In the calcipotriol-treated group the expression of K10 and K15 increased and the expression of K6 decreased significantly. No changes were seen for the marker beta-1 integrin. In the calcitriol-treated group none of the markers changed significantly. A tendency towards significance was seen for the changes in the expression of K6 and K15 in favour of calcipotriol.. Both calcitriol and calcipotriol gave a significant improvement in clinical scores. However, treatment with calcipotriol resulted in a normalization of K6, K10 and K15, whereas treatment with calcitriol did not. Comparison of both treatments showed a tendency towards significance for the above-mentioned markers for calcipotriol only.

Topics: Administration, Topical; Calcitriol; Calcium Channel Agonists; Cell Differentiation; Cell Proliferation; Dermatologic Agents; Double-Blind Method; Epidermal Growth Factor; Humans; Immunohistochemistry; Keratins; Ointments; Psoriasis; Treatment Outcome

Several reports have indicated that the combination of calcipotriol ointment and potent or ultrapotent corticosteroids are more effective and better tolerated, as compared to the monotherapies. The aim of the present study was to find out the effect of combination of calcipotriol ointment once daily and betamethasone dipropionate ointment once daily vs. the effect of twice-daily applications of each of the two treatments as monotherapy during a four-week treatment period. Seven patients with chronic plaque psoriasis were included for treatment with the three treatment schedules. Biopsies were taken before treatment and after four weeks of treatment, and markers for epidermal proliferation (Ki-67) and epidermal differentiation (keratin-10) were studied using a quantitative image analysis, and T-cell subsets in epidermis and dermis (CD4, CD8, CD25, CD45RO, CD45RA, CD94, CD161, and CD2) were studied using immunohistochemical scoring. The most impressive clinical result was reached with the combination. Calcipotriol proved to have a major effect on the proliferation marker Ki-67 and differentiation marker keratin-10, whereas the effect on T-cell subsets was more selective with major reductions of CD45RO(+) and CD8(+) T cells. In contrast, the effect of betamethasone dipropionate on the epidermis was restricted to a normalization of differentiation with a highly significant increase of keratin-10 positive epidermal surface without a significant effect on Ki-67 positive nuclei, and the effect on T-cell subsets was restricted to a reduction of natural killer T-cell receptors designated by CD94 and CD161 in the epidermis. The combination of the two treatments did not affect the proliferation marker Ki-67 and keratinization marker keratin-10, beyond the effect of calcipotriol monotherapy. However, the combination had a profound effect on, virtually, all T-cell subsets, beyond the effect of the monotherapies. It is concluded that the action spectra of calcipotriol and betamethasone on the psoriatic plaque are different and that the combination has effects on T-cell subsets, beyond the addition of the effects of monotherapies.

Topics: Anti-Inflammatory Agents; Antigens, CD; Betamethasone; Calcitriol; Cell Division; Dermis; Drug Therapy, Combination; Epidermis; Humans; Keratins; Ointments; Psoriasis; Skin; T-Lymphocyte Subsets; T-Lymphocytes

Vitamin D3 analogues are a first-line treatment of chronic plaque psoriasis, but so far, comparative clinical studies on calcipotriol and calcitriol ointment are sparse, and in particular no comparative studies are available on cell biological effects of these compounds in vivo. Using flow cytometric assessment, we investigated whether these compounds had different effects on the composition and DNA synthesis of epidermal cell populations responsible for the psoriatic phenotype. For 8 weeks, 20 patients with psoriasis vulgaris were treated twice daily with calcipotriol and calcitriol ointment in a left/right comparative study. Before and after treatment, clinical assessment of target lesions was performed, together with flow cytometric analysis of epidermal subpopulations with respect to keratin (K) 10, K6, vimentin and DNA distribution. Treatment with each compound resulted in a substantial clinical improvement, a reduction of the K10-K6- population and an increase of the K10+K6- population. A correlation was found between the clinical response of calcipotriol and the K10+K6- population, and the clinical response of calcitriol and the K10+K6- population, as well as the percentage of cells in the S, G2 and M phase of the cell cycle within the K10-K6- population, suggesting that the analogues have a different preference for affecting the K10+K6- pool (committed differentiated cells) or affecting the K10-K6- pool (basal cells).

Topics: Adult; Aged; Calcitriol; Cell Cycle; Dermatologic Agents; Drug Administration Schedule; Female; Flow Cytometry; Humans; Keratins; Male; Middle Aged; Ointments; Psoriasis; Regression Analysis; Vimentin

Calcipotriol and corticosteroids are established topical antipsoriatics. Previous studies have shown that combined therapy with calcipotriol and betamethasone dipropionate was more effective than monotherapy. In the present study, a recently developed combination product of calcipotriol and betamethasone dipropionate was compared with both monotherapies and the vehicle.. Twenty-five psoriatic patients were treated twice daily with the combination product, monotherapy or vehicle during 4 weeks. Skin biopsies, taken before and after treatment, were analysed using a multi-parameter flow cytometric method. Parameters of inflammation (vimentin-positive cells), normal differentiation (keratin-10-positive cells) and proliferation (cells in SG(2)M-phase) were assessed.. Flow cytometric analysis showed that the combination product turned out to be more effective in reducing inflammation compared with the other treatments. Restoration of normal differentiation was more advanced in patients treated with the combination product or betamethasone dipropionate compared to the vehicle. The highest number of normally differentiated cells was seen after use of the combination product. All treatments, except for the vehicle, decreased hyperproliferation. CONCLUSIONS This study shows that the combination product is a valuable new approach to the treatment of psoriasis.

Topics: Adult; Aged; Betamethasone; Calcitriol; Cell Cycle; Cell Division; Dermatologic Agents; Double-Blind Method; Drug Combinations; Female; Flow Cytometry; Humans; Keratin-10; Keratins; Male; Middle Aged; Psoriasis; Skin; Vimentin

Hydrocolloid (HCD) dressings enhance the efficacy of topical corticosteroids.. We wanted to evaluate the effect of calcipotriol ointment under an HCD dressing in the treatment of psoriatic plaques.. In 9 psoriatic patients, we cleared one plaque using this approach and took biopsies at start, clearance and relapse. Clinical and immunohistochemical validation was assessed.. After an average treatment of 3.6 weeks, each lesion had cleared (apart from some residual erythema). The average remission period was 8 weeks. During this treatment, the number of cycling epidermal cells (Ki-67-positive nuclei) and the expression of keratin 14 and keratin 16 had decreased substantially. In biopsies taken from the skin immediately adjacent to the relapsing lesion, these markers remained reduced which indicated the prolonged effect of calcipotriol on epidermal differentiation.. It is speculated that combination therapy of calcipotriol with treatments with a different mode of action such as photo(chemo)therapy, corticosteroids and cyclosporine might be worthwhile.

Topics: Adult; Aged; Antigens, Nuclear; Apoptosis; Calcitriol; Colloids; Dermatitis, Irritant; Dermatologic Agents; ErbB Receptors; Female; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Keratin-14; Keratins; Ki-67 Antigen; Male; Middle Aged; Nuclear Proteins; Occlusive Dressings; Ointments; Patient Dropouts; Psoriasis; Skin; Treatment Outcome

To examine the effects of topical therapy with Mahonia aquifolium on the expression of pathogenetically relevant molecules in psoriatic skin by immunohistochemistry.. Prospective-randomized, half-side comparison study with subsequent immunohistochemical assessment of biopsies.. The study areas were treated with Mahonia aquifolium ointment 3( daily and with dithranol in rising concentrations 1( daily, respectively. Biopsies of lesional skin from the test areas were carried out in 49 patients a) prior to therapy and b) 4 weeks after the start of therapy. Immunohistochemical stainings were performed with the following monoclonal antibodies: anti-ICAM-1, -CD3, -HLA-DR, -keratin 6, -keratin 16, -Ki-67. Evaluation of staining was made by two independent examiners using established semiquantitative scores.. Marked staining with all of the cited monoclonal antibodies was observed in the lesional skin prior to therapy. After 4 weeks of therapy there was a marked reduction in the expressions of ICAM-1, CD 3, HLA-DR and keratin 6 and 16. There were significantly greater reductions of ICAM-1, CD3, and HLA-DR at sites treated with dithranol. The expression of Ki-67 was not reduced by either therapy.. These results indicate efficacy of Mahonia aquifolium and dithranol in psoriatic skin both on cellular cutaneous immune mechanisms and on the hyperproliferation of keratinocytes. The effect of dithranol appears to be more potent than that of Mahonia aquifolium.

Topics: Administration, Topical; Anthralin; Anti-Inflammatory Agents; CD3 Complex; HLA-DR Antigens; Humans; Intercellular Adhesion Molecule-1; Keratins; Ki-67 Antigen; Ointments; Plants, Medicinal; Prospective Studies; Psoriasis; Skin

It is a well-established fact that very potent corticosteroids are highly effective in the treatment of psoriatic plaques, and that the addition of a hydrocolloid occlusive dressing (HCD) enhances its efficacy. It is known that topical steroids induce normal differentiation and diminish hyperproliferation during treatment of psoriatic plaques. These changes are reflected by an increase in keratin 10 (K10) and a decrease in keratin 6 (K6), respectively. However, the influence of class IV corticosteroids under HCD on K10 and K6 subpopulations has never been studied.. The present study was performed to study quantitatively the influence of a class IV topical steroid under HCD on the dynamics of K10 and K6 subpopulations.. In the present study, we treated moderately severe stable psoriatic plaques with clobetasol-17-propionate under HCD until clearance was achieved. We took biopsies prior to treatment, after clearance and at relapse. Using flow cytometry, we studied the dynamics of K10 and K6 subpopulations.. Prior to treatment, 41.8% of all cells expressed K6. After treatment-induced clearance, this proportion decreased to 1.2%, which is below the normal range. At relapse, pre-treatment levels were observed again. A trend to an increasing number of basal cells and an increase in the proliferative activity of these basal cells at relapse compared to the stable situation prior to treatment was observed.. We conclude that clobetasol under hydrocolloid occlusion induces virtually a total block of proliferation of the basal cell population and decreases hyperproliferation-associated keratins dramatically. Furthermore, based upon epidermal cell characteristics, we conclude that a rebound phenomenon occurs following discontinuation of therapy with clobetasol under hydrocolloid occlusion.

Topics: Administration, Topical; Anti-Inflammatory Agents; Clobetasol; Colloids; Flow Cytometry; Glucocorticoids; Humans; Keratin-10; Keratins; Occlusive Dressings; Psoriasis; Recurrence; Skin; Time Factors; Treatment Outcome

During the last decade, novel analogues of 1alpha,25-dihydroxy vitamin D3 have been developed for the treatment of psoriasis. Recently, the efficacy of short-term treatment with the novel derivative tacalcitol (1alpha,24-dihydroxy vitamin D3) has been documented. However, data on the long-term effect of tacalcitol on psoriatic skin are sparse. In this study, we assessed the cell characteristics of the psoriatic epidermis after treatment with tacalcitol for up to 24 weeks. We investigated how long-term treatment with tacalcitol modulates the percentages of differentiated keratinocytes, inflammation cells and basal keratinocytes, and the percentage of cells in the SG2M phase in the basal cell population. From 11 patients who were treated with tacalcitol for up to 18 months, we obtained single-cell suspensions of a representative psoriatic lesion after 0, 8, 12, 18 and 24 weeks of treatment. A Psoriasis Area and Severity Index was performed at each visit as well. Cell suspensions were stained with markers for inflammation (Vim3B4), differentiation (RKSE60) and proliferation (TO-PRO-3 iodide) and analysed flow cytometrically. Clinically, patients improved significantly after 8 weeks of treatment. This clinical effect was preserved for the rest of the period of treatment with no further significant improvement. Proliferative activity also decreased significantly after 8 weeks of treatment. Proliferation did not show further significant decreases or habituation after 12, 18 and 24 weeks. For inflammation, no statistically reliable trends could be seen. Differentiation improved significantly after 8 weeks of treatment, but decreased again significantly after 12 weeks. In the period from 12 to 24 weeks, no further significant change was observed. We conclude that tacalcitol is an effective antipsoriatic drug. Prolonged treatment with tacalcitol will generally maintain improvement at the level reached after 8 weeks. Owing to the beneficial effect on both clinical state and proliferation, tacalcitol is likely to be an adequate maintenance therapy.

Topics: Anti-Inflammatory Agents; Cell Differentiation; Cell Division; Dihydroxycholecalciferols; Drug Administration Schedule; Epidermis; Female; Flow Cytometry; Follow-Up Studies; Humans; Keratins; Male; Middle Aged; Psoriasis; Treatment Outcome

Recently we have described two novel markers for disturbed epidermal differentiation, which are strongly upregulated in psoriatic epidermis: skin-derived antileukoproteinase (SKALP) and epidermal fatty acid-binding protein (E-FABP). No data are available on the kinetics of SKALP and E-FABP expression in vivo and the relation with epidermal growth and differentiation. We used treatment of lesional psoriatic skin with topical steroid as a model to correlate the expression pattern of SKALP and E-FABP with known cell biological events during regression of the psoriatic lesion. Expression of these markers was studied using immunohistochemistry and Northern blot analysis. After 4 weeks of treatment a substantial clinical improvement was induced by the topical steroid, whereas no significant improvement had occurred at the placebo-treated sides. The expression of SKALP following treatment with steroid was nearly undetectable both at the protein and mRNA level. Mitotic activity, as measured by Ki-67 staining, and cytokeratin 16 expression were downregulated to normal levels in the steroid-treated epidermis. In contrast, although there was a marked decrease of E-FABP mRNA, the staining pattern for E-FABP at the protein level was not affected. After 4 weeks of treatment with steroid the complete suprabasal compartment remained positive, even after considerable clinical improvement of the lesion. We conclude that SKALP and cytokeratin 16 are markers that are downregulated even before complete macroscopic clearance of the lesion. The kinetics of E-FABP expression is distinct from the other molecules and lags behind the clinical signs of psoriasis.

Topics: Administration, Topical; Adult; Aged; Anti-Inflammatory Agents; Biomarkers; Blotting, Northern; Carrier Proteins; Double-Blind Method; Epidermis; Fatty Acid-Binding Protein 7; Fatty Acid-Binding Proteins; Female; Humans; Hydrocortisone; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Middle Aged; Myelin P2 Protein; Neoplasm Proteins; Phenotype; Proteinase Inhibitory Proteins, Secretory; Proteins; Psoriasis; RNA, Messenger; Tumor Suppressor Proteins

Objective comparison of different antipsoriatic therapies requires quantitative assessment of disease severity. However, clinical assessment with the widely used Psoriasis Area and Severity Index (PASI) introduces inaccuracy. An alternative is the quantitative analysis of different epidermal cell parameters using multiparameter flow cytometry. Our aim in the present study was to compare the clinical and flow cytometric approach to monitor disease activity and to evaluate antipsoriatic efficacy. Clinical scores for erythema, induration and scaling were assessed and biopsies for flow cytometric analysis were obtained from the psoriatic plaques of 89 patients before and after treatment with different therapeutic regimens consisting of vitamin D3 analogues and corticosteroids. In total, 219 epidermal cell suspensions were analysed using triple-labelling, with the simultaneous staining of markers for epidermal proliferation (DNA dye TO-PRO-3), differentiation (antikeratin 10), and inflammation (antivimentin). Correlation analysis was performed on 166 paired values obtained from 83 patients. A highly significant correlation was observed between erythema and the percentage of vimentin-positive cells, between scaling and the percentage of keratin 10 positive keratinocytes, and between induration and the number of basal keratinocytes in the S- and G2M phase, when all 166 biopsies were assessed. The correlation remained in the same range if the analysis was restricted to the 83 pretreatment biopsies. In contrast to the clinical scores, the flow cytometric analysis permitted a clear separation between the antiproliferative and anti-inflammatory or keratinization-enhancing effects of antipsoriatic treatment. The vitamin D3 analogues proved to exert a mainly antiproliferative effect. The combination of calcipotriol and a topical corticosteroid improved all cell biological markers substantially, and clobetasol monotherapy had a powerful effect on these markers. In conclusion, multiparameter flow cytometry has been shown to be a sensitive tool to evaluate the growth inhibiting, anti-inflammatory and keratinization-enhancing effects of antipsoriatic therapies.

Topics: Biomarkers; Cell Cycle; Epidermis; Erythema; Flow Cytometry; Humans; Keratins; Psoriasis; Severity of Illness Index; Treatment Outcome; Vimentin

The mechanisms of action of urea-containing ointments in the treatment of eczema, ichthyosis and psoriasis are only partly known and related to proteolysis and keratinolysis. In this study, we have examined the effects of topical urea on epidermal proliferation and differentiation in 10 patients with psoriasis. Plaque type lesions were treated for 2 weeks with an ointment containing 10% urea, with the vehicle alone, or left untreated. Clinical score, hydration of the stratum corneum, transepidermal water loss (TEWL), and immunohistochemical studies were performed. Epidermal proliferation was assessed using the Ki-S3 proliferation-associated nuclear antigen. For epidermal differentiation antibodies against involucrin and against keratins CK 5, 6, 17 and CK 1, 5, 10, 14 were used. The patients showed a reduction of the clinical score (> 50%), a 2-fold increase in stratum corneum hydration (p < 0.01), and a small decrease in TEWL (N.S.) on the urea- treated compared to the untreated site. Light microscopy studies revealed a 29% reduction in epidermal thickness (p < 0.01); epidermal proliferation was decreased by 51% (p < 0.005). The altered expression of involucrin and of cytokeratins (reduction of CK 5, 1 and 10 and induction of CK 6 and 17) was partially reversed. The ointment base also improved psoriasis, but urea was significantly better than the vehicle (urea: 40% reduction in epidermal proliferation vs. vehicle). In summary, these studies show that urea influences epidermal proliferation and differentiation in psoriasis.

Topics: Administration, Topical; Adolescent; Cell Differentiation; Cell Division; Double-Blind Method; Epidermis; Humans; Immunohistochemistry; Keratins; Ointments; Protein Precursors; Psoriasis; Urea

Calcipotriol and corticosteroids, two therapy modalities frequently prescribed in the treatment of psoriasis, are often used in combination. The aim of the present study was to determine whether the cell biological response pattern of concurrent use of calcipotriol and corticosteroids is different from calcipotriol monotherapy. Forty patients with chronic plaque psoriasis were divided at random in four parallel groups and treated for 8 weeks with: (1) calcipotriol cream (50 micrograms/g once daily); (2) calcipotriol cream twice daily; (3) calcipotriol and clobetasone 17-butyrate (0.5 mg/g) creams; and (4) calcipotriol and betamethasone 17-valerate (1 mg/g) creams. Before and after treatment keratotome biopsies were taken and single cell suspensions prepared for flow cytometric analysis. Flow cytometric multiparameter quantification of markers for proliferation (TO-PRO-3), differentiation (antikeratin 10) and inflammation (antivimentin) was used to evaluate all four therapy modalities. A statistically significant decrease of the percentage of basal cells in S- and G2M-phase (proliferation) was obtained with all therapy modalities, except for calcipotriol monotherapy applied once daily. A significant reduction of the number of vimentin-positive cells (non-keratinocytes) was observed following combined treatment with calcipotriol and clobetasone butyrate. In contrast, monotherapy with calcipotriol had virtually no effect on the number of vimentin-positive cells. It can be concluded that: (i) calcipotriol monotherapy, applied once daily was less antiproliferative compared with twice daily applications of calcipotriol or the combined treatment with corticosteroids and that (ii) the combination of calcipotriol and corticosteroids proved to have a marked effect on the percentage of non-keratinocytes, in contrast to the modest effect of calcipotriol.

Topics: Administration, Topical; Anti-Inflammatory Agents; Betamethasone Valerate; Calcitriol; Cell Division; Clobetasol; Dermatologic Agents; DNA; Drug Administration Schedule; Drug Therapy, Combination; Epidermis; Flow Cytometry; Glucocorticoids; Humans; Keratins; Psoriasis; Vimentin

The clinical efficacy and tolerability of the vitamin D3 analogues calcitriol, calcipotriol and 1 alpha, 24 dihydroxyvitamin D3 in the treatment of psoriasis have been assessed in various clinical studies. In vitro and in vivo investigations have shown interference of these compounds with epidermal growth, keratinisation and inflammation. In this study we quantified the in vivo cell biological effects during treatment of psoriatic plaques with 1 alpha, 24 dihydroxyvitamin D3. By using a flow cytometric triple labelling procedure, we could discriminate different epidermal subpopulations, permitting precise assessment of epidermal cell cycle kinetics. Twenty patients with plaque-type psoriasis were treated in a double-blind placebo-controlled left-right comparative study with 1 alpha, 24 dihydroxyvitamin D3 ointment (4 micrograms/g applied once daily) for 8 weeks. Epidermal cell suspensions prepared from keratotome biopsies taken before and after treatment were stained with TO-PRO-3 iodide (a new DNA fluorochrome) and monoclonal antibodies against keratin 10 (as a marker for differentiation) and vimentin (as a marker for inflammation), simultaneously. The flow cytometric analyses showed a significant decrease of proliferating basal keratinocytes in verum-treated lesions, whereas such a decrease was not observed in placebo-treated lesions. The amount of keratin 10-positive keratinocytes increased and the presence of vimentin-positive cells decreased in cell suspensions derived from both verum- and placebo-treated lesions, but these effects were not significant. We conclude that multiparameter flow cytometry promises to be an adequate approach to assess the interference of antipsoriatic treatments with cutaneous inflammation, epidermal proliferation and keratinisation. Topical 1 alpha, 24 dihydroxyvitamin D3 seems to exert its in vivo antipsoriatic effect mainly through an inhibition of epidermal growth.

Topics: Administration, Cutaneous; Adult; Aged; Biopsy; Cell Cycle; Cell Differentiation; Cell Division; Cells, Cultured; Dermatologic Agents; Dihydroxycholecalciferols; DNA; Double-Blind Method; Epidermis; Female; Flow Cytometry; Fluorescent Dyes; Humans; Inflammation; Keratins; Male; Middle Aged; Placebos; Psoriasis; Vimentin

Corticosteroids are important in the treatment of inflammatory dermatoses, such as psoriasis. They have anti-inflammatory, anti-proliferative and immunosuppressive effects. In this study, the effect of budesonide on proliferation, inflammatory cells and cytokines in psoriasis was investigated. In order to elucidate the time course of the different effects of corticosteroid treatment in psoriasis, six patients were treated for 3 weeks with budesonide 0.025% ointment (Preferid), and biopsies were studied immunohistochemically, before treatment and after 1 and 3 weeks of treatment. Clinical scores together with staining with antibodies indicating proliferation, keratin 16, keratin 10, T-lymphocytes, monocytes, polymorphonuclear leukocytes, Langerhans cells, interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-alpha (TNF-alpha), and intercellular adhesion molecule-1 (ICAM-1) were performed. 'Psoriasis area' and 'severity index' (PASI) scores were significantly reduced after 1 week and 3 weeks of treatment. Epidermal hyperproliferation (Ki-67 binding) and suprabasal keratin 16 (Ks8.12) expression decreased within 1 week, while keratin 10 (RKSE60) expression did not change. Five out of 6 patients showed cytokine levels (IL-1alpha, IL-6, IL-8, and TNF-alpha; detected immunohistochemically) in the normal range, while 1 patient had highly increased cytokine levels. In this patient, cytokine levels decreased during treatment. In 4 patients, showing high dermal ICAM-1 expression before treatment, a consistent reduction of ICAM-1 on endothelial cells was observed. The inflammatory infiltrate (T-lymphocytes (T11), monocytes/macrophages (WT14), polymorphonuclear leukocytes (PMN, anti-elastase)) was reduced to some extent after 3 weeks. The number of Langerhans cells (OKT6) did not change. These results indicate that the psoriatic lesions, although clinically comparable, show interindividual differences in cytokine expression. Corticosteroid treatment for 1-3 weeks improves clinical scores and hyperproliferation. Cytokine levels are reduced during steroid treatment in the patient who showed high levels before treatment. To suppress the infiltrate entirely, longer steroid treatment is probably necessary. This may explain the relapse seen after short term corticosteroid therapy.

Topics: Administration, Topical; Adult; Aged; Anti-Inflammatory Agents; Biomarkers; Budesonide; Cell Division; Cytokines; Female; Glucocorticoids; Humans; Immunohistochemistry; Inflammation; Inflammation Mediators; Keratins; Male; Middle Aged; Pregnenediones; Psoriasis; Time Factors

Oral retinoids have been widely used in psoriasis, but topical forms have been ineffective or irritating.. Our purpose was to determine the clinical and molecular effects of a new topical retinoid, AGN 190168, on psoriasis.. Seven patients with psoriasis were treated for 2 weeks with topical retinoid and 2 weeks with vehicle. Two control subjects with psoriasis were treated for 2 weeks with vehicle alone. Biopsy specimens from normal skin as well as from untreated and treated psoriatic lesions were compared by immunohistochemical analysis. Differentiation and inflammatory markers were studied.. Clinical improvement was seen in all seven patients after 2 weeks of treatment. Improvement was still present, but not significant, after 2 additional weeks of vehicle application. Histologic examination showed a return to a more normal morphology in four of seven biopsy specimens, which correlated with filaggrin expression. There was a diminution in the precocious expression of keratinocyte transglutaminase, keratin 16, and involucrin, as well as a decrease in epidermal growth factor receptor and in the number of cells expressing intercellular adhesion molecule type 1 and HLA-DR.. Clinical and histologic improvements were seen in psoriasis in association with the topical application of AGN 190168 at 2 weeks, including decreased inflammation and restoration of normal epidermal differentiation. Small patient numbers and the possibility that the changes were related to clinical improvement alone and not the topical agent preclude definitive conclusions.

Topics: Administration, Cutaneous; Adult; Antigens, CD; Biopsy; Cell Adhesion Molecules; Double-Blind Method; Epidermis; ErbB Receptors; Female; Filaggrin Proteins; Follow-Up Studies; HLA-DR Antigens; Humans; Immunohistochemistry; Intercellular Adhesion Molecule-1; Intermediate Filament Proteins; Keratinocytes; Keratins; Male; Nicotinic Acids; Pilot Projects; Prospective Studies; Protein Precursors; Psoriasis; Retinoids; Severity of Illness Index; Skin; Transglutaminases

The aim of the present study was to discover to what extent 1,24(OH)2D3 ointment (tacalcitol; 4 micrograms/g) can modulate epidermal proliferation and keratinization, and several aspects of inflammation. Ten patients with psoriasis vulgaris were included in a placebo-controlled, double-blind study, using 1,24(OH)2D3 ointment (4 micrograms/g). Before, and after 8 weeks of treatment, punch biopsies were taken from lesions treated with the active agent and placebo-treated lesions. An immunohistochemical study was carried out using monoclonal antibodies against the hyperproliferation-associated keratin 16, against cycling nuclei, filaggrin, involucrin, T lymphocytes, Langerhans cells, CD14 and polymorphonuclear leucocytes (PMN). The Wilcoxon test for matched pairs was used for statistical analysis of results. The biopsies from the lesions treated with the active agent showed a statistically significant change towards normalization of all aspects of inflammation studied, and of epidermal proliferation and keratinization, but there did not appear to be any effect on Langerhans cells. The only parameter which showed a significant alteration in the placebo-treated lesions was the number of cycling nuclei in the epidermis (P < or = 0.02). However, the biopsies from the plaques treated with the active agent showed a greater decrease of cycling cells (decrease: Mactive = 70, Mplacebo = 53) and a lower P-value (< or = 0.01). We therefore conclude that at the cell biological level 1,24(OH)2D3 ointment (4 micrograms/g) has a substantial effect on several cell types, with regard to inflammation, epidermal proliferation and keratinization, with the exception of Langerhans cells.

Topics: Cell Division; Dihydroxycholecalciferols; Double-Blind Method; Epidermis; Filaggrin Proteins; Humans; Immunohistochemistry; Inflammation; Keratins; Psoriasis

Changes in epidermal immunocytes and cytokeratins were investigated during treatment of psoriasis with calcipotriol and betamethasone valerate. Skin biopsies were obtained from 10 subjects on each treatment from lesional and non-lesional skin at baseline, and from treated lesions after 4 weeks. In each subject, changes in expression of cytokeratins K5, K10 and K16, and changes in epidermal immunocyte counts were assessed. Responses were compared with a separate histological parameter of improvement, epidermal thickness. Both treatments produced a marked normalization of cytokeratins. The reduction of K16 expression was similar on each treatment and correlated significantly with reduction in epidermal thickness. Expression of both K5 and K10 improved less than thickness with betamethasone valerate but more than thickness with calcipotriol, although these differences did not reach statistical significance. With calcipotriol there was an increase in K5 and K10 responses with increasing response of epidermal thickness, which was not seen with betamethasone valerate. T6+ cells, HLA-DR+ dendritic cells and T lymphocytes were all reduced by betamethasone valerate. There was a remarkable similarity in the level of normalization between cell types and also between cellular response and reduction in thickness. Calcipotriol produced a similar consistent reduction in cell numbers and in thickness, with the exception of T6+ cells which increased in some subjects during treatment. Only in subjects in whom thickness had virtually returned to normal was there a marked decrease in T6+ cells.

Topics: Adult; Aged; Antigen-Presenting Cells; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Betamethasone; Calcitriol; Dendritic Cells; Epidermis; Female; Humans; Keratins; Male; Middle Aged; Psoriasis; T-Lymphocytes

Antipsoriatic agents have been shown to decrease skin levels of arachidonic acid and its metabolites including 12-monohydroxy-eicosatetranoic acid (12-HETE), and leukotriene B4 (LTB4). In addition, specific systemic and topical lipoxygenase inhibitors have been reported to be effective in the treatment of psoriasis. The objective of this study was to investigate the effect of a potent oral leukotriene biosynthesis inhibitor (MK886) in patients with chronic plaque psoriasis. Clinical response together with the changes of LTB4 levels in lesional skin biopsy specimens, and urinary leukotriene E4 (LTE4) excretion were evaluated. In addition, markers of inflammation, proliferation and keratinization were studied immunohistochemically. No change in clinical scores or lesional LTB4 levels were observed with a 10 1/3-day course of MK886. A statistically significant reduction in urinary LTE4 excretion was observed: mean LTE4 (ng/h) were 5.14 before treatment and 1.51 on day 11 with MK886; and 7.55 before treatment and 6.57 on day 11 with placebo treatment. Epidermal accumulation of polymorphonuclear leukocytes (PMN) tended to diminish in the MK886 treatment group. These results indicate that although a reduction (greater than 70%) in urinary LTE4 excretion was found, and a slight decrease of epidermal PMN accumulation was observed, no correlative changes in clinical scores or LTB4 levels in skin lesion were found with a short course of MK886.

Topics: Administration, Oral; Adult; Aged; Biomarkers; Biopsy; Dermatitis; Double-Blind Method; Female; Humans; Immunohistochemistry; Indoles; Keratins; Leukotriene Antagonists; Leukotriene B4; Male; Middle Aged; Psoriasis; Skin

Twenty patients with psoriasis were treated with the vitamin D3 analogue MC903 and betamethasone ointment in a double-blind trial with a left-right comparison. In addition to the clinical severity scores, Ks8.12 binding which detects keratin 16 expression and the DNA synthesis were quantified using flow cytometry. Both markers decreased significantly with treatment, but remained above the normal range even in those who had total clearance of the lesions. Treatment with MC903 with regard to Ks8.12 binding was significantly better than with betamethasone.

Topics: Adult; Aged; Antibodies, Monoclonal; Betamethasone; Calcitriol; DNA; Double-Blind Method; Female; Fluorescent Antibody Technique; Humans; Keratins; Male; Middle Aged; Psoriasis; Skin

The relevance of salicylic acid in dithranol creams was evaluated in a double-blind study. Patients with chronic plaque psoriasis were treated using a short-contact schedule for dithranol on an outpatient basis. A left-right comparison was carried out between sites treated with either dithranol with 2% salicylic acid (D + S) or dithranol in the same base without salicylic acid (D-S). Clinical results were evaluated once a week using the psoriasis area severity index. In order to quantify the improvement, flow cytometric measurements were done using the monoclonal antibody Ks8.12, recognizing keratin 16 in normal and lesional epidermis. Simultaneously, relative DNA content was quantified which previously was described as a useful method to monitor a therapeutic effect. Both PASI scores and Ks8.12 binding decreased after 6 weeks treatment with D + S and D-S. However, percentages of cells in SG2M phases did not show a significant change. No significant difference was observed between sites treated with either D + S or D-S. Therefore we conclude that the addition of salicylic acid in a concentration of 2% does not enhance the efficacy of dithranol creams and we confirm that Ks8.12 is a useful quantitative marker for therapeutic efficacy.

Topics: Adolescent; Adult; Anthralin; DNA; Double-Blind Method; Erythema; Female; Flow Cytometry; Humans; Keratins; Male; Middle Aged; Ointments; Psoriasis; Randomized Controlled Trials as Topic; Salicylates; Salicylic Acid; Skin

Orally administered retinoids are synthetic derivatives of vitamin A. This new group of drugs (not yet available for general use in the United States) has been effective in experimental trials for treatment of a wide range of skin diseases. The current status of two of these drugs, isotretinoin (13-cis-retinoic acid) and etretinate (Ro 10-9359), is herein reviewed.

Topics: Acne Vulgaris; Administration, Oral; Child; Clinical Trials as Topic; Facial Dermatoses; Female; Humans; Isomerism; Isotretinoin; Keratins; Keratitis; Neoplasms; Psoriasis; Skin Diseases; Tretinoin; Xerostomia

Twenty patients with symmetric lesions of psoriasis were asked to use a polyester fiber sponge in addition to their other anti-psoriatic medications. The sponge was used to remove scales from those lesions on the left side only. The hypothesis was that removal of scales would enhance penetration by topical corticosteroid and/or coal tar preparations. At the end of four weeks, as well as at each weekly evaluation, all patients showed a statistically significant difference between the treated and control sides. In 70 percent of the subjects, results were graded good to excellent when the treated side was compared with the control side after four weeks' evaluation.

Topics: Administration, Topical; Adolescent; Adult; Aged; Child; Coal Tar; Female; Fluocinolone Acetonide; Fluocinonide; Humans; Keratins; Male; Middle Aged; Ointments; Polyesters; Psoriasis; Triamcinolone

Topics: Adult; Biopsy; Clinical Trials as Topic; Feces; Female; Humans; Keratins; Male; Placebos; Psoriasis; Skin; Sulfates; Zinc

Emerging evidence suggests that signaling through the C3a anaphylatoxin receptor (C3aR) protects against various inflammation-related diseases. However, the role of C3aR in psoriasis remains unknown. The purpose of this study was to investigate the possible protective role of C3aR in psoriasis and to explore the underlying molecular mechanisms. We initially found that the psoriatic epidermis exhibited significantly decreased C3aR expression. C3aR showed protective roles in mouse models of imiquimod (IMQ)- and interleukin-23-induced psoriasis. Furthermore, increased epidermal thickness and keratin 6 (K6), K16, and K17 expression occurred in the ears and backs of C3aR

Topics: Anaphylatoxins; Animals; Cell Proliferation; Disease Models, Animal; Keratin-16; Keratin-17; Keratin-6; Keratinocytes; Keratins; Mice; Psoriasis; Receptors, G-Protein-Coupled; Skin

Increased keratinocyte proliferation occurs in the skin of psoriatic patients and is supposed to play a role in the pathogenesis of this disorder. Compounds interfering with keratinocyte proliferation could be useful in the management of psoriatic patients.. To investigate whether albendazole, an anti-helmintic drug that regulates epithelial cell function in various systems, inhibits keratinocyte proliferation in models of psoriasis.. Aldara-treated mice received daily topical application of albendazole. Keratinocyte proliferation and keratin (K) 6 and K16 expression were evaluated by immunohistochemistry and Western blotting and inflammatory cells/mediators were analysed by immunohistochemistry and real-time PCR. In human keratinocytes (HEKa and HaCaT) treated with albendazole, cell cycle and proliferation, keratins and cell cycle-associated factors were evaluated by flow cytometry, colorimetric assay and Western blotting respectively.. Aldara-treated mice given albendazole exhibited reduced epidermal thickness, decreased number of proliferating keratinocytes and K6/K16 expression. Reduction of CD3- and Ly6G-positive cells in the skin of albendazole-treated mice associated with inhibition of IL-6, TNF-α, IL-1β, IL-17A, IL-36, CCL17, CXCL1, CXCL2 and CXCL5 expression. Treatment of keratinocytes with albendazole reduced K6/K16 expression and reversibly inhibited cell growth by promoting accumulation of cells in S-phase. This phenomenon was accompanied by down-regulation of CDC25A, a phosphatase regulating progression of cell cycle through S-phase, and PKR-dependent hyper-phosphorylation of eIF2α, an inhibitor of CDC25 translation. In Aldara-treated mice, albendazole activated PKR, enhanced eIF2α phosphorylation and reduced CDC25A expression.. Data show that albendazole inhibits keratinocyte proliferation and exerts therapeutic effect in a murine model of psoriasis.

Topics: Albendazole; Animals; cdc25 Phosphatases; Cell Line; Cell Proliferation; Cytokines; Dermatologic Agents; Disease Models, Animal; eIF-2 Kinase; Eukaryotic Initiation Factor-2; Humans; Imiquimod; Inflammation Mediators; Keratinocytes; Keratins; Male; Mice, Inbred C57BL; Phosphorylation; Psoriasis; S Phase Cell Cycle Checkpoints; Signal Transduction; Skin

Interleukin 17A (IL-17A), mainly produced by the T helper subclass Th17, plays a key role in the psoriatic plaque formation and progression. The clinical effectiveness of anti-IL-17A agents is documented, but the early and specific mechanisms of their protection are not identified yet. The challenge of the present study is to investigate the possible reversal exerted by a specific anti-IL-17A agent on the psoriatic events induced by IL-17A in a three-dimensional organotypic model of normal human skin. Bioptic skin fragments obtained after aesthetic surgery of healthy women (n=5) were incubated with i) IL-17A biological inhibitor (anti-IL-17A), ii) IL-17A, iii) a combination of IL-17A and its specific IL-17A biological inhibitor (COMBO). A Control group was in parallel cultured and incubation lasted for 24 and 48 h epidermal-side-up at the air-liquid interface. All subjects were represented in all experimental groups at all considered time-points. Keratinocyte proliferation and the presence of epidermal Langerhans cells were quantitatively estimated. In parallel with transmission electron microscopy analysis, immunofluorescence studies for the epidermal distribution of keratin (K)10, K14, K16, K17, filaggrin/occludin, Toll-like Receptor 4, and Nuclear Factor kB were performed. IL-17A inhibited cell proliferation and induced K17 expression, while samples incubated with the anti-IL-17A agent were comparable to controls. In the COMBO group the IL-17A-induced effects were almost completely reverted. Our study, for the first time, elucidates the most specific psoriatic cellular events that can be partially affected or completely reverted by a specific anti-IL-17A agent during the early phases of the plaque onset and progression. On the whole, this work contributes to expand the knowledge of the psoriatic tableau.

Topics: Adult; Antibodies, Monoclonal; Female; Filaggrin Proteins; Humans; Interleukin-17; Keratins; Langerhans Cells; NF-kappa B p50 Subunit; Occludin; Psoriasis; S100 Proteins; Skin; Toll-Like Receptor 4; Young Adult

Psoriasis is a common inflammatory skin disease characterized by hyperproliferation of epidermal keratinocytes, however, there is still no curative therapy for psoriasis. Paeoniflorin (PF), a Chinese herbal medicine, has shown anti-inflammatory effects.. We aimed to illustrate the effect and associated mechanism of PF on keratinocyte proliferation using the IMQ-induced psoriasis mouse model.. The anti-psoriatic effect of PF in vivo and in vitro was assessed by western blot, RT-PCR, immunofluorescence, cell counting kit-8 and haematoxylin/eosin staining.. In vivo, epidermal thickness, dermal infiltrated lymphocytes and the level of several antimicrobial peptides, pro-inflammatory cytokines, and K17 were significantly reduced in mice with topical application of PF. In vitro, PF inhibited the proliferation of HaCaT cells in a dose-dependent manner, down-regulated K17 expression, and suppressed NF-kappaB activation.. Our findings indicate that PF may inhibit the proliferation of keratinocytes by targeting K17, suggesting that PF might be a potential target in the treatment of psoriasis.

Topics: Administration, Cutaneous; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Proliferation; Cytokines; Down-Regulation; Female; Glucosides; HaCaT Cells; Humans; Imiquimod; Keratin-17; Keratinocytes; Keratins; Mice; Monoterpenes; NF-kappa B; Psoriasis; Signal Transduction

Curcuma was the dried rhizomes of Curcuma kwangsiensis S.G. Lee et C.F. Liang (Chinese name: e zhu), have been used in China for thousands of years. There are some reports have shown that curcumin, the major component of curcuma, has a good curative effect on psoriasis, but the mechanism is still unknown, so the present study was designed to investigate the effect of curcuma's extraction on psoriasis-like mouse, and to explore the mechanisms of therapy. First, we observed that curcuma's extractions effect on mitosis of mouse vaginal epithelial cells; then making psoriasis like model and measuring the score of skin damage on days 7 and 14; finally, we observed the expression of immune factors (CK14, CK16, CK17, PCNA, TLR-2, TLR-4, TLR-9) in propranolol induced psoriasis like rats. Curcuma's extraction prohibited the mitosis of mouse vaginal epithelial cells; curcuma's extractions have a significantly efficacy and dose dependent inhibition on imiquimod induced psoriasis like rats; and the expression of immune factors (CK14, CK16, CK17, PCNA, TLR-2, TLR-4, TLR-9) was decreasing in the curcuma's extraction treated groups compared with normal groups. Our research proved that curcuma's extractions have a significantly efficacy on psoriasis like rats, thus, curcuma's extractions can be a potential novel treatment for psoriasis. Furthermore, the expression of immune factors was decreasing after treatment with curcuma's extraction suggest us cytokines has strong relation with the mechanism of therapy for psoriasis. Our results contribute towards validation of curcuma in the treatment of psoriasis and other joint disorders.

Topics: Animals; Curcuma; Dermatologic Agents; Disease Models, Animal; Dose-Response Relationship, Drug; Epithelial Cells; Female; Guinea Pigs; Imiquimod; Keratins; Male; Mice; Mitosis; Plant Extracts; Proliferating Cell Nuclear Antigen; Propranolol; Psoriasis; Rhizome; Skin; Time Factors; Toll-Like Receptors; Vagina

Psoriasis is a common immune-mediated disease resulting from altered cross-talk between keratinocytes and immune cells. Previous transcriptomic studies have identified thousands of deregulated genes in psoriasis skin; however, the transcriptomic changes confined to the epidermal compartment remained poorly characterized. The aim of this study was to characterize the transcriptomic landscape of psoriatic keratinocytes, using sorted CD45neg epidermal cells. Genes with functions in innate immunity, type I interferon response, cell cycle and keratinization were enriched among deregulated genes in psoriatic keratinocytes. Gene set enrichment analysis indicated the dominance of interleukin (IL)-22/IL-17A signatures in the epidermal psoriasis-signature. A set of deregulated genes overlapped with psoriasis-associated genetic regions, suggesting that genetic variations affecting gene expression in keratinocytes contribute to susceptibility to psoriasis. Several psoriasis-susceptibility genes, which were previously believed to be expressed preferentially or exclusively in immune cells, were identified as having altered expression in psoriatic keratinocytes. These results highlight the role of keratinocytes in the pathogenesis of psoriasis, and indicate that both genetic factors and an inflammatory microenvironment contribute to epidermal alterations in psoriasis.

Topics: Adult; Aged; Case-Control Studies; Cell Cycle; Cellular Microenvironment; Epidermis; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Regulatory Networks; Humans; Immunity, Innate; Interleukin-17; Interleukin-22; Interleukins; Keratinocytes; Keratins; Male; Middle Aged; Oligonucleotide Array Sequence Analysis; Psoriasis; Signal Transduction; Transcriptome; Young Adult

Topics: Aminoquinolines; Animals; Apoptosis; Biomarkers; Cell Line; Cell Proliferation; Disease Models, Animal; Down-Regulation; Female; Healthy Volunteers; Humans; Imiquimod; Keratin-17; Keratinocytes; Keratins; Male; Mice; Mice, Inbred BALB C; Microscopy, Electron, Transmission; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Primary Cell Culture; Psoriasis; Radiation Dosage; Signal Transduction; Skin; STAT3 Transcription Factor; Treatment Outcome; Ultraviolet Therapy

Mutations in keratin proteins have been vastly associated with a wide array of genodermatoses; however, mutations of keratins in psoriasis have not been fully investigated. The main aim of the current research was to identify the mutation in K14, K10, K16, and K17 genes in two stages of psoriasis patients.. Ninety-six psoriatic skin biopsies were collected. mRNA transcript of K14, K10, K16, and K17 was prepared, amplified, and sequenced. Sanger sequences of all keratins were further validated for mutational analysis using Mutation Surveyor and Alamut Visual. Then, in silico analysis of protein stability and protein and gene expression of all keratins was performed and validated.. Out of 44 mutations, about 75% of keratins are highly pathogenic and deleterious. Remaining 25% mutations are less pathogenic and tolerated in nature. In these 33 deleterious mutations were immensely found to decrease keratin protein stability. We also found a correlation between keratin and Psoriasis Area and Severity Index score which added that alteration in keratin gene in skin causes severity of psoriasis.. We strongly concluded that acanthosis and abnormal terminal differentiation was mainly due to the mutation in epidermal keratins. In turn, disease severity and relapsing of psoriasis are mainly due to the mutation of hyperproliferative keratins. These novel keratin mutations in psoriatic epidermis might be one of the causative factors for psoriasis.

Topics: Acanthosis Nigricans; Adolescent; Adult; Aged; Biopsy; Cell Differentiation; Cell Proliferation; DNA Mutational Analysis; Epidermis; Female; Humans; Keratins; Keratins, Type I; Male; Middle Aged; Mutation; Protein Stability; Psoriasis; Severity of Illness Index; Skin; Young Adult

Psoriasis is a common chronic dermatosis characterized by keratinocyte hyperproliferation accompanied by inflammatory reactions. Pathological changes upset the balance between keratinocyte proliferation, differentiation, and death in psoriatic lesions, suggesting that molecules with topical anti-inflammatory, anti-proliferation and anti-angiogenesis abilities may be useful for its treatment. The flavonoid astilbin is the major active component extracted from the rhizome of Smilax glabra, which has been widely used in China to treat inflammatory and autoimmune diseases. Here, we investigate the potential of astilbin as a treatment for psoriasis. We reveal that astilbin inhibits the growth of HaCaT keratinocytes. Detailed study shows that astilbin leads to S phase arrest of the cell cycle by induction of p53 and p21 and activated-AMPK. Additionally, astilbin induced keratinocyte differentiation correlated with suppression of keratin 5 (KRT5) and KRT14 proteins (the markers of epidermal basal layer) and induction KRT1 and KRT10 proteins (occurring in the upper layers). Moreover, astilbin regulates the expression of VEGF in human HaCaT keratinocytes. These results suggest that astilbin may be a promising agent for psoriasis treatment.

Topics: Anti-Inflammatory Agents; Cell Differentiation; Cell Line; Cell Proliferation; China; Cyclin-Dependent Kinase Inhibitor p21; Flavonols; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; Rhizome; S Phase; Tumor Suppressor Protein p53

Psoriasis is a chronic inflammatory skin disease severely affecting patients' physical and psychological well-being. Aberrant keratin expression in psoriasis plays a crucial role in keratinocyte dysfunction and disease development. Little is yet known about the mechanism of this keratin dysregulation. VEGF (Vascular endothelial growth factor) is significantly elevated in psoriatic patients and VEGFRs are detected on keratinocytes, leading to our hypothesis that this keratin dysregulation may be regulated by VEGF. In this study, we showed that VEGFR2 was overexpressed in psoriatic epidermis and was correlated with K (keratin) 6, K16&K17 upregulation and K1&K10 downregulation. VEGF increased both mRNA and protein levels of K6, K16&K17 and decreased those of K1&K10 in NHEKs (normal human epidermal keratinocytes). Further we identified activation of STAT3, ERK1/2, and p38 pathways in VEGF-treated NHEKs. Using specific pathway antagonists and siRNAs we found that VEGF induced K6, K16&K17 via these three pathways and reduced K1&K10 via ERK1/2. Finally, VEGF-induced aberrant keratin expression pattern and epidermal thickening were confirmed in a VEGF-local-injection mouse model. Collectively, we demonstrated that VEGF was associated with aberrant keratin expression pattern in psoriasis and provided a new insight into the role of VEGF in psoriasis pathogenesis, indicating VEGF as a potential therapeutic target.

Topics: Adolescent; Adult; Animals; Cells, Cultured; Child; Epidermis; Female; Gene Expression Profiling; Gene Expression Regulation; Humans; Keratins; Male; Mice; Mice, Inbred BALB C; Psoriasis; Skin; Vascular Endothelial Growth Factor A; Young Adult

Topics: Animals; Climatic Processes; Denial, Psychological; Female; Fraud; Humans; Keratins; Male; Motivation; Peptide Hydrolases; Psoriasis; Sepharose; Skin; Space Flight; United States National Aeronautics and Space Administration

Topics: Animals; Climatic Processes; Denial, Psychological; Female; Fraud; Humans; Keratins; Male; Motivation; Peptide Hydrolases; Psoriasis; Sepharose; Skin; Space Flight; United States National Aeronautics and Space Administration

Topics: Cell Lineage; Cell Proliferation; Cells, Cultured; Collagen Type IV; Cytoplasm; Epidermis; Gene Expression Regulation; Humans; Keratinocytes; Keratins; MicroRNAs; Psoriasis; Stem Cells; Transfection

Keratin-specific immune responses in tonsils may be associated with the pathogenesis of pustulosis palmaris et plantaris (PPP). Evaluation of keratin-specific immune responses in tonsils might be useful to predict the effectiveness of tonsillectomy for patients with PPP.. The aim of the present study was to clarify the role of keratin-specific immune responses in the pathogenesis of PPP in tonsils. It has been reported that anti-keratin antibodies in serum were higher in patients with PPP and decreased after tonsillectomy, indicating that anti-keratin antibodies might be generated in tonsils.. In order to demonstrate the presence of keratin-specific immune responses in tonsils, the numbers of keratin-specific antibody-forming cells (AFCs) in tonsillar and peripheral blood lymphocytes were examined by enzyme-linked immunospot assay. The prognosis of PPP was compared after tonsillectomy.. The numbers of keratin-specific IgM and IgG AFCs in tonsils and of IgG AFCs in peripheral blood were significantly increased in patients with PPP. The numbers of keratin-specific IgG AFCs in peripheral blood correlated positively with tonsil and serum IgG antibodies specific to keratin. Our data show that a good prognosis in patients with PPP depended on the numbers of keratin-specific IgG and IgM AFCs in peripheral blood and the levels of keratin-specific IgG antibodies in serum being significantly decreased 6 months after tonsillectomy.

Topics: Adolescent; Adult; Antibody-Producing Cells; Case-Control Studies; Female; Humans; Keratins; Male; Middle Aged; Palatine Tonsil; Prognosis; Psoriasis; Tonsillectomy; Young Adult

Keratins are cytoskeletal intermediate filament proteins that are increasingly being recognised for their diverse cellular functions. Here we report the consequences of germ line inactivation of Keratin 76 (Krt76) in mice. Homozygous disruption of this epidermally expressed gene causes neonatal skin flaking, hyperpigmentation, inflammation, impaired wound healing, and death prior to 12 weeks of age. We show that this phenotype is associated with functionally defective tight junctions that are characterised by mislocalization of the integral protein CLDN1. We further demonstrate that KRT76 interacts with CLDN1 and propose that this interaction is necessary to correctly position CLDN1 in tight junctions. The mislocalization of CLDN1 has been associated in various dermopathies, including the inflammatory disease, psoriasis. These observations establish a previously unknown connection between the intermediate filament cytoskeleton network and tight junctions and showcase Krt76 null mice as a possible model to study aberrant tight junction driven skin diseases.

Topics: Animals; Claudin-1; Cytoskeleton; Epidermis; Humans; Intermediate Filaments; Keratinocytes; Keratins; Mice; Psoriasis; Skin Diseases; Tight Junctions

Cells of the epidermis renew constantly from germinal layer stem cells. Although epithelial cell differentiation has been studied in great detail and the role of Wnt signaling in this process is well described, the contribution of epidermal Wnt secretion in epithelial cell homeostasis remains poorly understood. To analyze the role of Wnt proteins in this process, we created a conditional knockout allele of the Wnt cargo receptor Evi/Gpr177/Wntless and studied mice that lacked Evi expression in the epidermis. We found that K14-Cre, Evi-LOF mice lost their hair during the first hair cycle, showing a reddish skin with impaired skin barrier function. Expression profiling of mutant and wild-type skin revealed up-regulation of inflammation-associated genes. Furthermore, we found that Evi expression in psoriatic skin biopsies is down-regulated, suggesting that Evi-deficient mice developed skin lesions that resemble human psoriasis. Immune cell infiltration was detected in Evi-LOF skin. Interestingly, an age-dependent depletion of dendritic epidermal T cells (DETCs) and an infiltration of γδ(low) T cells in Evi mutant epidermis was observed. Collectively, the described inflammatory skin phenotype in Evi-deficient mice revealed an essential role of Wnt secretion in maintaining normal skin homeostasis by enabling a balanced epidermal-dermal cross talk, which affects immune cell recruitment and DETC survival.

Topics: Animals; CD3 Complex; Cell Proliferation; Chronic Disease; Dendritic Cells; Dermatitis; Epidermis; Gene Deletion; Humans; Inflammation; Intracellular Signaling Peptides and Proteins; Keratinocytes; Keratins; Lymphocyte Activation; Mice; Mice, Transgenic; Neutrophil Infiltration; Phenotype; Psoriasis; Receptors, Antigen, T-Cell, gamma-delta; Receptors, G-Protein-Coupled; STAT3 Transcription Factor; T-Lymphocytes; Wnt Proteins

Ligands of the EGF family regulate autocrine keratinocyte proliferation, and IL-1 family cytokines orchestrate epithelial defense responses. Although members of both families are overexpressed in wound healing and psoriasis, their roles in regulating the innate immune functions of keratinocytes remain incompletely explored. Using sensitive assays, we found significant increases of heparin-binding EGF-like growth factor, transforming growth factor-α, and amphiregulin mRNA and protein in lesional psoriasis compared with uninvolved or control skin. In normal human keratinocyte (NHK) monolayers, EGFR ligands were ineffective in inducing DEFB4, S100A7, and CCL20 mRNAs and human β-defensin (hBD)-2 peptide. Combined with IL-1α, however, EGFR ligands provoked 250 × more DEFB4 and CCL20 and a 9-fold rise in S100A7 mRNA relative to the EGFR ligand alone. This synergy was also reflected in secreted hBD-2 protein, both from NHK and reconstituted human epidermis. Keratinocyte differentiation was critical for these responses, as postconfluent NHK yielded mRNA and protein levels an order of magnitude greater than subconfluent cells. Differentiation also influenced signal transduction, with subconfluent cells using NF-κB and postconfluent cells using EGFR, MEK1/2, and p38. We propose that EGFR ligands are important modifiers of IL-1 activity, synergizing with IL-1 to stimulate epidermal production of hBD-2, S100A7, and CCL20, three of the most upregulated transcripts in psoriatic plaques.

Topics: Adolescent; Adult; Aged; beta-Defensins; Biopsy; Calcium; Cell Count; Cell Differentiation; Cells, Cultured; Chemokine CCL20; ErbB Receptors; Humans; Immunity, Innate; Interleukin-1; Keratinocytes; Keratins; Middle Aged; Psoriasis; S100 Calcium Binding Protein A7; S100 Proteins; Signal Transduction; Young Adult

Retinoids (RA) have been used as therapeutic agents for numerous skin diseases, from psoriasis to acne and wrinkles. While RA is known to inhibit keratinocyte differentiation, the molecular effects of RA in epidermis have not been comprehensively defined. To identify the transcriptional targets of RA in primary human epidermal keratinocytes, we compared the transcriptional profiles of cells grown in the presence or absence of all-trans retinoic acid for 1, 4, 24, 48, and 72 h, using large DNA microarrays. As expected, RA suppresses the protein markers of cornification; however the genes responsible for biosynthesis of epidermal lipids, long-chain fatty acids, cholesterol, and sphingolipids, are also suppressed. Importantly, the pathways of RA synthesis, esterification and metabolism are activated by RA; therefore, RA regulates its own bioavailability. Unexpectedly, RA regulates many genes associated with the cell cycle and programmed cell death. This led us to reveal novel effects of RA on keratinocyte proliferation and apoptosis. The response to RA is very fast: 315 genes were regulated already after 1 h. More than one-third of RA-regulated genes function in signal transduction and regulation of transcription. Using in silico analysis, we identified a set of over-represented transcription factor binding sites in the RA-regulated genes. Many psoriasis-related genes are regulated by RA, some induced, others suppressed. These results comprehensively document the transcriptional changes caused by RA in keratinocytes, add new insights into the molecular mechanism influenced by RA in the epidermis and demonstrate the hypothesis-generating power of DNA microarray analysis.

Topics: Cell Differentiation; Cells, Cultured; Epidermal Cells; Gene Expression Profiling; Gene Expression Regulation; Humans; Keratinocytes; Keratins; Keratolytic Agents; Lipid Metabolism; Multigene Family; Oligonucleotide Array Sequence Analysis; Promoter Regions, Genetic; Psoriasis; Receptors, Retinoic Acid; Transcription, Genetic; Tretinoin

Clear-cell acanthoma (CCA) has been reported to be a benign epidermal neoplasm; however, several authors have suggested alternative differentiation as well as other nosologic categories, including a reactive dermatosis. Fourteen CCAs, ten tricholemmomas, and seven cases of psoriasis were reviewed with conventional microscopy, periodic acid-Schiff stains, and immunohistochemical stains. Twelve of fourteen (86%) CCAs were associated with underlying or adjacent conditions. The CCAs stained immunohistochemically in a pattern similar to normal epidermis and psoriasis. Tricholemmomas stained in a distinctly different pattern with MNF116 and NGFR/p75. These cases demonstrate CCA in settings that reflect chronic inflammation, primarily scars and stasis dermatitis, and with an immunophenotype that parallels psoriasis. These findings support the contention that CCA does not show outer follicular sheath (tricholemmal) differentiation. Furthermore, these cases lend additional support to the contention that CCA represents a psoriasiform reaction pattern, which, in appropriately taken biopsies, usually has a demonstrable associated condition. Nonetheless, the precise nosology of this phenomenon has yet to be elucidated completely.

Topics: Acanthoma; Adult; Aged; Aged, 80 and over; Cicatrix; Dermatitis; Epidermis; Female; Hair Follicle; Hidradenitis Suppurativa; Humans; Hyperplasia; Keratins; Keratosis, Seborrheic; Male; Middle Aged; Molecular Weight; Neoplasms, Basal Cell; Nerve Tissue Proteins; Psoriasis; Receptors, Nerve Growth Factor; Skin; Skin Neoplasms

The effect of the established antipsoriatic treatment with topical calcipotriol (with a maximum of 100 g per week) in addition to systemic treatment with alefacept, a new biological agent for psoriasis, on epidermal cell populations in the psoriatic lesion was investigated using a combination of the Zenon labelling technique and microscopic image analysis. Epidermal cell populations were measured quantitatively with this sensitive method.. Frozen sections of non-treated psoriatic epidermis and psoriatic epidermis treated with either alefacept intramuscular or alefacept intramuscular in combination with topical calcipotriol for 12 weeks were compared immunohistochemically. Antibodies against keratin 6, 10 and 15 were labelled with the Zenon technique, whereas antibodies against the Ki-67 antigen and beta-1 integrin were covalently Fluorescein Isothiocyanate (FITC)-labelled. Using image analysis, these markers were measured in the epidermis in a standardized manner.. Treatment of psoriasis with alefacept resulted in a good clinical response in several patients and in a normalization of epidermal expression of the immunohistochemical parameters for differentiation and proliferation. The addition of topical calcipotriol resulted in a faster clinical improvement with a similar overall clinical response and a similar response of epidermal cell populations as compared to treatment with alefacept monotherapy after 12 weeks of treatment. This study also suggests that the appearance of keratin 15 has a predictive value for the duration of remission. It can be concluded that the addition of a low-dose calcipotriol treatment does not contribute to the clinical efficacy of alefacept, both at the clinical level and with respect to markers for epidermal proliferation and differentiation.

Topics: Administration, Topical; Alefacept; Biomarkers; Calcitriol; Dermatologic Agents; Epidermis; Female; Fluorescent Dyes; Humans; Immunohistochemistry; Integrin beta1; Keratins; Ki-67 Antigen; Male; Middle Aged; Psoriasis; Recombinant Fusion Proteins; Staining and Labeling; Treatment Outcome

Identification of critical autoantigenic T-cell epitopes is key to developing antigen-based therapies for autoimmune diseases, including psoriasis. Our previous work demonstrated that 3 peptides on keratin 17 are able to stimulate peripheral blood lymphocytes of HLA-DRB1*07-positive patients with psoriasis and to serve as immunodominant T-cell epitopes.. We sought to determine antagonistic altered peptide ligands to psoriatic T cells with a down-modulatory effect in inhibiting keratinocyte proliferation.. Psoriatic altered peptide ligands were generated by single alanine residue substitutions at a critical T-cell receptor contact residue position. Antagonistic altered peptide ligands were identified by suppression screening of psoriatic T-cell activation and keratinocyte proliferation.. Altered peptide ligands 119R and 355L can inhibit psoriatic T-cell activation more effectively than other altered peptide ligands, especially 355L, with inhibition of T-cell proliferation and the secretion of interferon gamma and interleukin 2 in parallel with the up-regulation of interleukins 4 and 10 as well as transforming growth factor-beta. In coincubation assay, altered peptide ligands 119R and 355L can down-regulate the function of psoriatic T cells more effectively than wild-type epitopes solely, but less effectively than altered peptide ligands solely. In prepulse assay altered peptide ligand 119R can down-regulate the activation of psoriatic T cells more effectively than in coincubation but less effectively as compared with altered peptide ligand 119R only. Altered peptide ligand 355L was also shown to have a similar presentation. T-cell culture supernatants (1:100) from the concentrations (10 microg.mL(-1) and 100 microg.mL(-1) with 119R, 100 microg.mL(-1) with 355L) were more effective than the other ratios in inhibiting keratinocyte proliferation.. This study had a relatively small sample size (52 patients and 48 healthy controls).. Our findings show that the altered peptide ligands 119R (VAALEEANTELEVKI) and 355L (ENRYCVQASQIQGLI) are capable of inhibiting proliferative responses of psoriatic T cells and keratinocyte proliferation in vitro, at least, with enhanced helper T cell type 2 polarization. Thus, to our knowledge, this article is the first report of the demonstration of therapeutic activity of altered peptide ligands derived from keratin 17.

Topics: Cell Proliferation; Cells, Cultured; Humans; Keratinocytes; Keratins; Ligands; Peptides; Psoriasis; Receptors, Antigen, T-Cell; T-Lymphocytes

We report the use of non-invasive tape stripping to sample psoriatic lesional and non-lesional skin in 96 patients. The procedure was well tolerated with any discomfort described as mild; we did not observe any cases of Koebner phenomena at any non-lesional tape-stripped sites. Tape-harvested epidermis was extracted for RNA, which was profiled by semiquantitative reverse transcriptase-PCR. This analysis revealed that mRNAs for tumor necrosis factor alpha, IFNgamma, Krt-16, CD2, IL-23A, IL-12B, and vascular endothelial growth factor are overexpressed in the "average" psoriatic lesion in a majority of patients. In addition, 10 of these patients were biopsied at lesional and non-lesional sites and the expression data compared to tape-stripping data. This comparison shows that five of seven mRNA are more highly expressed in cells captured by tape stripping than biopsy, suggesting that the upper aspect of a lesion contains cells very active in the disease. The tape-harvesting data reveal that approximately 46% of lesions have at least one pathogenic mRNA within non-lesional skin limits. Data demonstrate that tape stripping reveals mRNA markers not detected in biopsy samples and thus the method may be a useful supplement to biopsy.

Topics: Biopsy; Humans; Keratin-16; Keratins; Psoriasis; RNA, Messenger; Skin; Tumor Necrosis Factor-alpha

The margin zone in spreading psoriatic lesions has frequently been used as a model to study the changes in epidermal proliferation, keratinization and inflammation during the transition from symptomless to lesional skin. However, the dynamics of the changes in the epidermal subpopulations-basal cells, transit amplifying cells and differentiated cells-have not been studied in the transition between symptomless and lesional skin.. To quantify in a dynamic model of the margin zone in psoriasis the characteristics of these subpopulations with respect to epidermal proliferation and differentiation.. From seven patients with active psoriasis, biopsies were taken from the distant uninvolved skin, outer margin, inner margin and centre of a spreading psoriatic plaque. Frozen sections were labelled immunofluorescently using direct immunofluorescence for Ki-67 and beta1 integrin and the Zenon labelling technique for keratin 6, 10 and 15. Digital photographs of the stained sections were quantitatively analysed.. In the distant uninvolved skin the expression of beta1 integrin was decreased and keratin 15 expression was lost. In this area suprabasal cells expressed beta1 integrin and in the outer margin suprabasal cells expressed Ki-67. From the outer to the inner margin of the psoriasis plaque, which coincided with the appearance of the clinical lesion, there was a significant change in the various markers. The patchy expression of keratin 6 in the inner margin became homogeneous in the centre of the psoriasis plaque and here was also coexpression of keratin 6 and keratin 10 in a single cell.. The present study provides additional evidence that the distant uninvolved skin has a prepsoriatic phenotype, which is the first step in a psoriatic cascade. The cascade between symptomless and lesional skin comprises first an abnormality in inflammation with involvement of beta1 integrin-dim cells (transit amplifying cells) subsequently eliciting an enlarged germinative compartment with increased recruitment of cycling epidermal cells and focal expression of proliferation-associated keratins, ultimately culminating in a more-or-less homogeneous epidermis with massive recruitment of cycling epidermal cells and proliferation-associated keratinization.

Topics: Cell Differentiation; Cell Proliferation; Epidermis; Fluorescent Antibody Technique, Direct; Humans; Image Processing, Computer-Assisted; Integrin beta1; Keratins; Ki-67 Antigen; Psoriasis

Avarol, a marine sesquiterpenoid hydroquinone, and 14 avarol derivatives have shown interesting anti-inflammatory properties in previous studies. In this study, avarol and derivatives were evaluated in high-throughput keratinocyte culture models using cytokeratin 10 and SKALP/Elafin expression as markers for respectively normal and psoriatic differentiation. Avarol and five of its derivatives (5, 10, 13, 14 and 15) were selected for further study. Only 10, 13, 14 and 15 were able to inhibit keratinocyte cell growth. Changes in expression levels of 22 genes were assessed by quantitative real time PCR (qPCR). From these genes, TNFalpha mRNA levels showed the strongest changes. For compound 13, 15 and dithranol (used as a model antipsoriatic drug), a dose-dependent downregulation of TNFalpha mRNA was found. The changes in TNFalpha mRNA were confirmed at the protein level for compound 13. Additionally, this compound was able to reduce also IL-8 and COX-2 mRNA levels and this effect was correlated with a reduction in COX-2 protein expression. The mechanism of action of this compound involves at least the inhibition of NF-kappaB-DNA binding activity. In conclusion, our high-throughput screening models in combination with quantitative assessment of changes in gene expression profiles identified the avarol derivative 13, a benzylamine derivative of avarol at the 4' position of benzoquinone ring, as an interesting anti-psoriatic drug candidate that inhibits keratinocyte cell growth and TNFalpha and COX-2 expression.

Topics: Animals; Antineoplastic Agents; Cell Differentiation; Cell Proliferation; Cells, Cultured; Cyclooxygenase 2; Drug Evaluation, Preclinical; Dysidea; Elafin; Enzyme-Linked Immunosorbent Assay; Humans; In Vitro Techniques; Interleukin-8; Keratinocytes; Keratins; Membrane Proteins; Psoriasis; RNA, Messenger; Sesquiterpenes; Tumor Necrosis Factor-alpha

Thermal balneotherapy with Comano's spa water (CW; Trentino, Italy) is used for psoriasis and other skin disorders but its mechanisms of action are mostly unknown. Previously, we showed that CW can interfere with the expression and secretion of various VEGF-A isoforms by cultured human psoriatic epidermal keratinocytes. In this study, confluent cultures of IL-6-hypersecreting keratino-cytes isolated from 6 psoriatic patients were exposed for 11-15 days to DMEM, the chemicals of which had been dissolved in either deionised water (DW-DMEM, controls) or CW (CW-DMEM, treated cells). As detected by means of immunocytochemistry, Western immunoblotting, and ELISA assays, the intracellular levels and secretion rates of IL-6 were drastically curtailed in the CW-DMEM-incubated keratinocytes and in their cell-conditioned media. A nearly maximal inhibition of IL-6 release had already been induced by a CW fraction in the DMEM as low as 25%. CW exposure also promptly, intensely, and persistently down-regulated the expression of cytokeratin-16 (CK-16), a marker associated with keratinocyte psoriatic phenotype. Hence, CW balneotherapy may beneficially affect the clinical manifestations of psoriasis via an attenuation of the local deregulation of several cytokines/chemokines, including IL-6 and VEGF-A isoforms, and of a concurrent, abnormal cell differentiation program entailing the expression, amongst other proteins, of CK-16.

Topics: Cells, Cultured; Humans; Interleukin-6; Italy; Keratinocytes; Keratins; Mineral Waters; Psoriasis

Our knowledge of the etiology and pathogenesis of skin diseases characterized by abnormal growth is relatively limited. Even more important, still very little is known of how epidermopoeisis is controlled in normal epidermis. There is no cure for skin diseases caused by abnormal growth control, such as psoriasis. Mechanisms are complex, additional models for epidermal growth and differentiation, and specific techniques to analyze these processes, are needed. Therefore, we have developed flow cytometric techniques to study epidermal growth over the past two decades. A prerequisite for accurate and reliable flow cytometric analysis is a high quality of epidermal single-cell suspensions. In this chapter, protocols are described for preparation of single-cell suspensions and protocols for the multiparameter flow cytometric analysis of growth behavior in normal and hyperproliferative epidermis.

Topics: Cell Differentiation; Cell Division; Flow Cytometry; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; Skin

In order to better characterize epidermal cell populations in psoriatic vs. normal skin, fluorescent immunohistochemical techniques were extended with a new labeling technique. The Zenon technique conjugates primary antibodies rapidly and quantitatively after which they are used in the same manner as covalently labeled primary antibodies. Digital microscopic images of epidermal expression of keratin 10 and keratin 6 (differentiation), Ki-67 antigen (proliferation), and keratin 15 and beta-1 integrin (basal layer) were analyzed in a standardized way. Co-expression of different proteins was demonstrated.. Sections of normal skin and psoriatic lesions were compared immunohistochemically. Antibodies against keratin 6, 10, and 15 were labeled with the Zenon technique. Antibodies against the Ki-67 antigen and beta-1 integrin were covalently fluorescein isothiocyanate-labeled. Using standardized image analysis, intensity and positive surface area of the different antibodies in the epidermis were measured.. The number of Ki-67-antigen positive cells was significantly increased in lesional psoriatic skin. Intensity and positive surface area of keratin 10 and beta-1 integrin were significantly decreased in comparison to normal epidermis. Differential expression of keratin 6 and keratin 15 was demonstrated.. Using Zenon technology and image analysis, a description of morphology, co-expression, and quantification of representative markers for epidermal cell populations is possible.

Topics: Biomarkers; Cell Count; Epidermis; Female; Fluorescent Dyes; Humans; Image Processing, Computer-Assisted; Integrin beta1; Keratins; Ki-67 Antigen; Male; Middle Aged; Psoriasis; Staining and Labeling

How the inflammatory response is initiated has been well defined but relatively little is known about how such responses are resolved. Here we show that the D6 chemokine receptor is involved in the post-inflammatory clearance of beta-chemokines from cutaneous sites. After induction of inflammation by phorbol esters, wild-type mice showed a transient inflammatory response. However, in D6-deficient mice, an excess concentration of residual chemokines caused a notable inflammatory pathology with similarities to human psoriasis. These results suggest that D6 is involved in the resolution of the cutaneous inflammatory response.

Topics: Animals; Chemokine CXCL2; Chemokine Receptor D6; Chemokines; Chemokines, CC; Dermatitis; Female; Histocytochemistry; Inflammation; Keratins; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Proliferating Cell Nuclear Antigen; Psoriasis; Receptors, CCR10; Receptors, Chemokine; Skin; von Willebrand Factor

Psoriasis is a T cell-mediated inflammatory skin disease. Recent evidence suggests that activated CD4+ helper T lymphocytes of the Th1 phenotype play an important role in the pathogenesis of the disease. For psoriatic autoreactive T cells, Keratin 17 is a major target antigen and an epitope containing ALEEAN sequence has been described, but other psoriasis-related epitopes are still unknown.. To identify the HLA DR B1*04, *07-restricted T cell epitopes on Keratin 17.. HLA DRB1*04, *07-restricted T cell epitope regions on Keratin 17 were predicted based on related software and internet servers. Keratin 17 gene was amplified from psoriatic epidermis and the proteins of the predicted epitope regions were expressed, identified and purified. T cells from psoriatic patients reacted in cultivation with peptide-major histocompatibility complex (p-MHC) compound, then the level of cell proliferation and the concentration of interferon-gamma in culture supernatant were detected. After the psoriasis-related epitope regions were narrowed down, the epitopes on them were predicted further. These epitopes were then expressed and validated by T cell response in vitro.. Four epitopes--S1 (118-132), S2 (169-183), S4 (323-337) and S4 (348-362) can stimulate the proliferation and interferon-gamma production of psoriatic T cells more effectively than other epitopes (p<0.01) and react weakly with the T cells from healthy volunteers (p>0.05).. Epitopes S1 (118-132), S2 (169-183), S4 (323-337) and S4 (348-362) are immunodominant DR B1-restricted T cell epitopes for psoriasis. Among them, S1 (118-132) contains the ALEEAN sequence while the others with different amino acid sequence have not been reported before. Further studies based on these peptides would provide a more complete understanding of the immunological basis of psoriasis.

Topics: Amino Acid Sequence; Autoimmunity; Base Sequence; Case-Control Studies; Cell Division; Cloning, Molecular; Epitopes; Gene Expression; HLA-DR Antigens; HLA-DRB1 Chains; Humans; Immunodominant Epitopes; Interferon-gamma; Keratins; Molecular Sequence Data; Psoriasis; T-Lymphocytes

A novel antibody labelling technique, the Zenon technique, was used in fluorescent immunohistochemistry for a better characterization of epidermal cell populations in a quantitative approach. With this technique, differences in proliferation and differentiation characteristics were shown between psoriatic and normal epidermis. The sensitivity of the method was investigated by assessing the effect of a mild topical treatment versus an emollient. Frozen sections of non-treated psoriatic epidermis and psoriatic epidermis treated once daily with either an emollient or betamethasone-17-valerate for only 2 weeks were compared immunohistochemically. Antibodies against keratin 6, 10 and 15 were labelled with the Zenon technique, whereas antibodies against the Ki-67 antigen and beta-1 integrin were covalently FITC-labelled. Using image analysis, these markers were measured in the epidermis in a standardized manner. Treatment of psoriasis with short-term topical steroid resulted towards normalization of Ki-67 antigen, beta-1 integrin, keratin 10 and keratin 6 expression, which are parameters for proliferation and differentiation. Although treatment with an emollient showed hardly any clinical response, changes towards a more normal phenotype could already be detected in several epidermal markers using this method.

Topics: Antibodies; Betamethasone Valerate; Drug Administration Schedule; Emollients; Female; Fluorescent Antibody Technique; Glucocorticoids; Humans; Integrin beta1; Keratins; Ki-67 Antigen; Male; Middle Aged; Predictive Value of Tests; Psoriasis; Sensitivity and Specificity; Skin

Monitoring dynamics of different cell populations in solid tissues using flow cytometry has several limitations. The interaction and changes in epidermal subpopulations in hyperproliferative skin disorders such as psoriasis, a very common chronic inflammatory skin disease, may, however, elucidate the role of different cell populations in the pathogenesis and the effect of therapy in these disorders. We describe a new, simple method for identifying and quantifying different epidermal subpopulations in hyperproliferative skin disorders and an in vivo model for epidermal hyperproliferation by using six-parameter assessment and three-color flow cytometry.. Epidermal single-cell suspensions were derived from normal human skin before and after standardized injury and from untreated and treated psoriatic lesions. Samples were stained with anti-cytokeratins 6 (K6) and 10 (K10), anti-vimentin, and 4',6-diamidino-2-phenylindole. With three-color flow cytometry, different epidermal subpopulations were further defined by six different parameters.. Correlations were found for mild psoriasis and K10(+)K6(-) cells and for moderate psoriasis and K10(-)K6(+) cells. Treatment of mild psoriasis resulted in a 70% increase of the K10(+)K6(-) cells and an 18% decrease of the K10(-)K6(-) cells, whereas treatment of more severe psoriasis resulted in a 77% increase of the K10(+)K6(-) cells and a 33% decrease of the K10(-)K6(+) cells. The tape-stripping model showed changes in suprabasal keratin expression before proliferative activity.. The present flow cytometric assay permits simultaneous quantification of epidermal differentiation, inflammation, proliferation, and hyperproliferation-associated keratinization and provides information on pathogenesis and therapy of hyperproliferative skin disorders.

Topics: Antibodies; Biopsy; Cell Division; Flow Cytometry; Humans; Keratins; Psoriasis; Reference Values; Regression Analysis; Skin

Psoriasis is recognized as a chronic inflammatory disease characterized by epidermal hyperproliferation. To identify psoriasis-related genes, we compared the mRNA populations of normal and psoriatic skin. We identified one gene, designated as cornifelin, which showed increased expression in psoriatic skin. Human cornifelin contains 112 amino acids and is expressed in the uterus, cervix, and skin. In situ hybridization analysis demonstrated the presence of human cornifelin in the granular cell layer of the epidermis. To investigate the function of cornifelin, we established a transgenic mouse line overexpressing human cornifelin. Using these mice, we have shown that cornifelin is directly or indirectly cross-linked to at least two other cornified envelope proteins, loricrin and involucrin, in vivo. Overexpression of human cornifelin correlated with decreased loricrin expression and increased involucrin expression in the transgenic mouse. However, abnormality of epidermal differentiation was not observed in the transgenic mouse.

Topics: Amino Acid Sequence; Animals; Base Sequence; Gene Expression; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Molecular Sequence Data; Protein Precursors; Psoriasis; Recombinant Proteins; RNA, Messenger; Sequence Homology, Amino Acid; Skin; Tissue Distribution

The expression of amphiregulin (AR) in the basal epidermis of transgenic mice [keratin 14 promoter AR gene (K14-ARGE)] has been previously shown to induce an early-onset and severe skin pathology, with many similarities to psoriasis. In this study, it is demonstrated that involucrin enhancer/promoter-dependent expression of human AR (INV-AR) in the suprabasal epidermis of transgenic mice also produces a cutaneous psoriasis-like phenotype. INV-AR mice possess a limited lifespan and scaling, papillomatous, erythematous skin with partial alopecia. INV-AR mouse histopathology also revealed epidermal hyperkeratosis, parakeratosis, acanthosis, and an exaggerated dermal vasculature. A dermal and epidermal infiltrate was also evident and consisted of both neutrophils and CD3(+) T lymphocytes. The histology of synovial joints in both the INV-AR mice and the K14-ARGE mice of our previous investigation was examined. The histologic examination revealed that 3-week-old INV-AR transgenic mice displayed normal knee joint histology, while 2- to 3-week-old K14-ARGE transgenic mice frequently displayed synovitis, as exemplified by the presence of a mixed leukocytic infiltration, increased vascularization, and enhanced deposition of fibrous matrix in the knee synovium. These results demonstrate that AR overexpression in both the basal and suprabasal epidermis of transgenic mice induces a phenotype that mimics cutaneous psoriasis, while basal AR expression is also associated with synovial inflammation, a precursor to the psoriasis-associated arthropathy, psoriatic arthritis. Collectively, the results implicate epidermal AR expression as a possible mediator of innate cutaneous immunity and epidermal proliferation and also as a potential trigger of both cutaneous psoriasis and psoriatic arthritis.

Topics: Age of Onset; Amino Acid Sequence; Amphiregulin; Animals; CD3 Complex; EGF Family of Proteins; Epidermis; Glycoproteins; Humans; Intercellular Signaling Peptides and Proteins; Keratin-14; Keratins; Knee Joint; Mice; Mice, Transgenic; Molecular Sequence Data; Promoter Regions, Genetic; Psoriasis; Severity of Illness Index; Synovitis; T-Lymphocytes

To search for autoantigens in psoriatic plaques, we screened cDNA libraries of plaque epidermis with psoriatic serum samples. This approach has been highly successful in identifying tumor antigens, but has not been widely applied to autoimmune disease. We identified 11 autoantigens including three with prominent reactivity and plausible disease relevance. These are keratin 13 (K13), heterogeneous nuclear ribonucleoprotein-A1 (hnRNP-A1), and a previously uncharacterized protein, FLJ00294. Serum antibody screening for these demonstrated reactivity in 40%, 38%, and 27% of psoriasis patients, respectively. Most positive samples reacted with all three, and we found that this was due to cross-reactivity among them. Enzyme-linked immunospot assay (ELISPOT) analysis of psoriatic peripheral blood T cells confirmed that these autoantigens are also recognized by T cells. This demonstrates that this is a feasible method to identify autoantigens in an autoimmune target tissue, and suggests that these antigens warrant further study in psoriasis. Furthermore, but peripheral blood of normal controls reacted to these autoantigens with essentially the same frequencies as patients, suggesting that psoriatics may have not only an immune system which is capable of reacting to certain autoantigens, but also to a skin immunoregulatory alteration which allows this normal reactivity to develop into abnormal inflammation.

Topics: Autoantigens; Cross Reactions; Gene Library; Genetic Testing; Heterogeneous Nuclear Ribonucleoprotein A1; Heterogeneous-Nuclear Ribonucleoprotein Group A-B; Humans; Keratins; Psoriasis

K16, a type I keratin, is upregulated in hyperproliferative states including psoriasis. It has been used as a marker of psoriasis and its expression is upregulated in relapsing psoriasis and downregulating in resolving. We evaluated non-lesional psoriatic skin for K16 expression.. Sixty-seven non-lesional and lesional skin samples from patients with psoriasis and normal skin from 19 non-psoriatic patients were studied by immunohistochemistry on frozen sections with K16.. Seventeen of 19 normal skin samples showed staining of basal cells in the deeper part of the rete ridges. Sixty-two non-lesional psoriatic skin samples showed intense basal staining of K16. Of the remaining five non-lesional samples, diffuse intense suprabasal staining in one, pan-epidermal staining in two, and no staining was seen in two samples. Suprabasal (37), diffuse (14), sandwich (12), and basal (3) pattern staining were seen in psoriatic skin. One psoriatic skin sample did not show any expression.. Our results demonstrate that K16 expression is also observed in non-lesional psoriatic skin and may serve as a marker of preclinical psoriasis.

Topics: Adolescent; Adult; Aged; Biomarkers; Female; Humans; Immunohistochemistry; Keratin-1; Keratins; Male; Middle Aged; Psoriasis; Skin

Antikeratin autoantibodies (AK auto Abs) are very important elements of the human immune system. To improve the outcome of studies on AK auto Abs, it is necessary to generate antikeratin human monoclonal antibodies. The purpose of present study was to isolate antikeratin human monoclonal antibodies by panning a phage antibody library. A semisynthetic phage antibody library with capacity of 4.0x10(8) members was previously constructed. Panning of the library was performed against human epidermal keratin extracted from psoriatic scales. At the last round of the panning, individual colonies were grown in culture for expression of phage antibodies. Their binding activities and specificities to keratin were determined by ELISA, and positive clones were analyzed by DNA fingerprinting. The selected clones were induced with IPTG to express soluble Fab fragments, which were further characterized by ELISA, immunohistochemistry and Western blotting. Finally, DNA sequencing of the variable regions was performed. A human antibody clone which was able to express soluble Fab fragments and recognize Mr 46,000 keratin (K17) was isolated. DNA sequencing demonstrated that the VH and VL of the antibody came from the human VH1 and Vkappa2 families, respectively. We conclude that phage antibody library technology is a powerful way to generate human monoclonal antibodies. The antikeratin antibody we isolated in the present study would be useful in the research on AK auto Abs.

Topics: Amino Acid Sequence; Antibodies, Monoclonal; Bacteriophages; Base Sequence; Blotting, Western; Cloning, Molecular; Enzyme-Linked Immunosorbent Assay; Epidermis; Gene Expression; Gene Library; Humans; Immunoenzyme Techniques; Immunoglobulin Fab Fragments; Keratins; Molecular Sequence Data; Psoriasis; Skin Neoplasms; Staining and Labeling

The association of psoriasis with Streptococcus pyogenes throat infections suggests a potential antigenic target for the T cells that are known to infiltrate psoriatic skin. Streptococcal M protein share an extensive sequence homology with the human epidermal keratins. Keratin 17 (K17), while being mostly absent from uninvolved skin, is up-regulated in psoriatic lesions. Consequentially, M-protein-primed T cells may recognize up-regulated keratin epitopes via molecular mimicry. Using in vitro lymphocyte culture and cytokine flow cytometry we demonstrate that HLA-Cw*0602(+) psoriasis patients had significant CD8(+) T cell interferon (IFN)-gamma responses to peptides from the K17 and M6 protein selected on the basis of sequence homology and predicted HLA-Cw*0602 binding. These responses were about 10 times more frequent in the skin-homing cutaneous lymphocyte-associated antigen-expressing (CLA(+)) subset of CD8(+) T cells. CD4(+) T cells showed only borderline responses. CLA(+) CD8(+) T cells from Cw6(+) non-psoriatic individuals responded to some M6 peptides but rarely to K17 peptides. Cw6(-) psoriasis patients showed a response that was intermediate between Cw6(+) patients and controls. These findings indicate that psoriatic individuals have CD8(+) T cells that recognize keratin self-antigens and that epitopes shared by streptococcal M proteins and human keratins may be targets for the CD8(+) T cells that infiltrate psoriatic skin lesions.

Topics: Antigens, Bacterial; Antigens, Differentiation, T-Lymphocyte; Antigens, Neoplasm; Bacterial Outer Membrane Proteins; Carrier Proteins; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Female; HLA-C Antigens; Humans; Interferon-gamma; Keratins; Leukocytes, Mononuclear; Male; Membrane Glycoproteins; Middle Aged; Peptides; Psoriasis; Receptors, Lymphocyte Homing; Sequence Homology, Amino Acid; Skin; Streptococcus pyogenes

The cytoskeleton in keratinocytes is a complex of highly homologous structural proteins derived from two families of type I and type II polypeptides. Keratin K2e is a type II polypeptide that is expressed in epidermis late in differentiation. Here we report the influence of keratinocyte activation, proliferation, and keratinization on K2e expression in samples of cutaneous and oral lesions. The normal expression of K2e in the upper spinous and granular layers of interfollicular epidermis is increased in keloid scars but showed distinct down-regulation in psoriasis and hypertrophic scars where keratinocytes are known to undergo activation. Unlike normal and psoriatic skin, K2e expression in hypertrophic and keloid scars began in the deepest suprabasal layer. In cutaneous basal and squamous cell carcinomas, K2e was absent in most tumor islands but the overlying epidermis showed strong expression. No significant K2e expression in nonkeratinized or keratinized oral epithelia, including buccal mucosa, lateral border of tongue and gingiva was detected. In oral lichen planus K2e expression was undetectable, but in benign keratoses of lingual mucosa induction of K2e along with K1 and K10 was observed. In mild-to-moderate oral dysplasia with orthokeratinization, K2e was highly expressed compared with parakeratinized areas but in severe dysplasia as well as in oral squamous cell carcinoma, K2e expression was undetectable. Taken together, the data suggest that K2e expression in skin is sensitive to keratinocyte activation but its up-regulation in oral lesions is a reflection of the degree of orthokeratinization.

Topics: Animals; Antibodies, Monoclonal; Base Sequence; Breast; Cell Division; DNA Primers; Epidermal Cells; Epidermis; Female; Gene Expression Regulation; Genetic Variation; Gingiva; Humans; In Situ Hybridization; Keloid; Keratinocytes; Keratins; Mice; Mouth Mucosa; Peptide Fragments; Polymerase Chain Reaction; Protein Isoforms; Psoriasis; Skin

Psoriasis has been recognized as an immunologically mediated inflammatory skin disease that has been associated with group A, beta-haemolytic streptococcal infections. Notably cross-reactive autoimmune mechanism, which is mediated by T cells reacting to epitopes that are common to streptococcal M-protein and keratin, has been proposed in psoriasis. In order to investigate this possibility, peptides corresponding to M-protein and human epidermal keratin, which share some amino acid sequence between them, were synthesized and tested for their ability to stimulate T-cells of patients with psoriasis. Among five cases examined, we isolated a CD4(+) T-cell line that recognized the type I keratin (K14)(p168-181) when it was presented by the patient's HLA-DR molecules from a single psoriatic patient, whose MHC allele was HLA-A2/A26, -B27/B16, -DR4/DR8, -DQ8. Further analysis disclosed that the critical peptide recognized by the T-cell line was 10-mer keratin(p171-180) (DLRNKILTAT). However, corresponding M6 protein with homology to K14 did not stimulate the T-cell response and no evidence for cross-reactivity was obtained. The K14-responsive T cell line produced IFN-gamma, but little IL-4 when stimulated with irradiated autologous PBMC pulsed with this peptide. Thus, the finding that human epidermal keratin peptide is immunogenic in a psoriasis patient may provide the evidence that T lymphocytes play an important role in the pathogenesis of psoriasis as an autoimmune disorder participated with Th1 like cells. However, the keratin-responsive T cell line was detected in only one of five cases of psoriasis examined, suggesting that such T cell line appears to be not so popular in psoriatic patients. No evidence for cross-reactivity to streptococcal M protein also suggests that the contribution of streptococci may simply be inducing proliferation of various repertoire of T cells (including K14-responsive T cells) possibly through a superantigen-dependent process.

Topics: Antigens, Bacterial; Bacterial Outer Membrane Proteins; Carrier Proteins; CD4-Positive T-Lymphocytes; Cell Line; Epitopes; Humans; Keratin-14; Keratins; Peptide Fragments; Psoriasis

Epithelial cell keratins make up the type I (K9-K20) and type II (K1-K8) intermediate filament proteins. In glandular epithelia, K8 becomes phosphorylated on S73 ((71)LLpSPL) in human cultured cells and tissues during stress, apoptosis, and mitosis. Of all known proteins, the context of the K8 S73 motif (LLS/TPL) is unique to type II keratins and is conserved in epidermal K5/K6, esophageal K4, and type II hair keratins, except that serine is replaced by threonine. Because knowledge regarding epidermal and esophageal keratin regulation is limited, we tested whether K4-K6 are phosphorylated on the LLTPL motif. K5 and K6 become phosphorylated in vitro on threonine by the stress-activated kinase p38. Site-specific anti-phosphokeratin antibodies to LLpTPL were generated, which demonstrated negligible basal K4-K6 phosphorylation. In contrast, treatment of primary keratinocytes and other cultured cells, and ex vivo skin and esophagus cultures, with serine/threonine phosphatase inhibitors causes a dramatic increase in K4-K6 LLpTPL phosphorylation. This phosphorylation is accompanied by keratin solubilization, filament reorganization, and collapse. K5/K6 LLTPL phosphorylation occurs in vivo during mitosis and apoptosis induced by UV light or anisomycin, and in human psoriatic skin and squamous cell carcinoma. In conclusion, type II keratins of proliferating epithelia undergo phosphorylation at a unique and conserved motif as part of physiological mitotic and stress-related signals.

Topics: Apoptosis; Cell Cycle; Cells, Cultured; Conserved Sequence; Humans; Keratinocytes; Keratins; Male; Mitosis; Organ Specificity; Phosphoproteins; Phosphorylation; Protein Isoforms; Psoriasis; Skin; Stress, Physiological

A structural, profile-based algorithm was used to identify interleukin 20 (IL-20), a novel IL-10 homolog. Chromosomal localization of IL-20 led to the discovery of an IL-10 family cytokine cluster. Overexpression of IL-20 in transgenic (TG) mice causes neonatal lethality with skin abnormalities including aberrant epidermal differentiation. Recombinant IL-20 protein stimulates a signal transduction pathway through STAT3 in a keratinocyte cell line, demonstrating a direct action of this ligand. An IL-20 receptor was identified as a heterodimer of two orphan class II cytokine receptor subunits. Both receptor subunits are expressed in skin and are dramatically upregulated in psoriatic skin. Taken together, these results demonstrate a role in epidermal function and psoriasis for IL-20, a novel cytokine identified solely by bioinformatics analysis.

Topics: Animals; Cell Line; Chromosome Mapping; Cloning, Molecular; Dimerization; DNA-Binding Proteins; Epidermis; Gene Expression; Humans; Interleukin-10; Interleukins; Keratinocytes; Keratins; Mice; Mice, Transgenic; Molecular Sequence Data; Psoriasis; Receptors, Cytokine; Sequence Homology, Amino Acid; STAT3 Transcription Factor; Trans-Activators; Up-Regulation

Epidermal changes overlying dermatofibromas (DFs) have been described as ranging from psoriasiform simple hyperplasia to basaloid hyperplasia sometimes morphologically indistinguishable from superficial basal cell carcinoma (BCC). To characterize epidermal hyperplasia overlying DFs and to determine its association with the disease process, we examined 30 cases of DF showing hyperplastic epidermis. We used nine immunohistochemical markers associated with keratinocyte proliferation or differentiation. In DFs, the dermal metallothionein (MT) expression and immunophenotypic changes with regard to epidermal differentiation varied depending on the stage of lesional evolution of the DFs. Immunostaining for epidermal growth factor receptor (EGFR), MT, and keratin 6 (K6) increased in simple hyperplastic epidermis (SHE) overlying DFs (n = 11), whereas it gradually diminished in basaloid hyperplastic epidermis (BHE) overlying DFs (n = 19). In SHE, there was a significant increase in K14 expression. Among 19 BHE cases, 12 showed premature expression of involucrin and delayed appearance of K1 along with aberrant expression of K14. Conversely, the remaining 7 BHE cases showed a pattern of involucrin and K1 similar to that of normal skin coinciding with decreased or absent dermal MT expression. Loricrin and filaggrin expression in all DFs was the same as that of normal skin. Based on the sparse positivity of Ki-67 in the hyperplastic epidermis overlying DFs, we found that the biologic ability of BHE and SHE was not apparent in the hyperproliferative state observed in psoriasis and BCC. These results suggest that the dermal fibrohistiocytic process may trigger the induction of SHE overlying DFs by an unknown mechanism and then mediate both the abnormal keratinocyte differentiation and the transformation of SHE to BHE through the evolution of the dermal lesions.

Topics: Biomarkers; Carcinoma, Basal Cell; Cell Differentiation; Cell Division; Epidermis; ErbB Receptors; Filaggrin Proteins; Histiocytoma, Benign Fibrous; Humans; Hyperplasia; Immunoenzyme Techniques; Keratinocytes; Keratins; Metallothionein; Precancerous Conditions; Psoriasis; Skin Neoplasms

Keratins are a group of cytoskeletal proteins that are found in human epidermis and other stratified squamous epithelia. Several different types of keratins have been described. Keratin 10 (K10) is a keratin that is expressed in well differentiated, suprabasal keratinocytes, and keratin 6 (K6) is a keratin which is associated with hyperproliferation. Psoriasis is a chronic inflammatory skin disease, and besides inflammation, disturbed differentiation and hyperproliferation are its hallmarks. In order to study the hyperproliferation associated keratinization in both well differentiated and poorly differentiated keratinocytes, and in order to assess the proliferative activity of all K10 and K6 subpopulations, simultaneous assessment of K6, K10, and DNA content is required. So far, a triple staining protocol had not been available. In the present study, we established a novel protocol for simultaneous measurement of K6, K10, and DNA content, which enables the characterization of the proliferative activity of several cellular subpopulations in epidermis. From 16 patients with psoriasis and from 15 healthy volunteers, punch biopsies were obtained. After preparation of single cell suspensions, cells were stained with the anti-keratin 10 IgG(1)-isotype monoclonal antibody RKSE60, with the anti-keratin 6 IgG(2a)-isotype monoclonal antibody LHK6B, and with the DNA fluorochrome TO-PRO-3 iodide. Isotype specific secondary antibodies conjugated with phycoerythrein (PE) and fluorescein isothiocyanate (FITC) were used as the second step in the staining procedure. Controls were measured omitting the primary antibodies, and gates were set in order to differentiate between the K10 and K6 subpopulations. Samples from both psoriatic patients and healthy volunteers were than measured. Owing to the IgG specificity of RKSE60 and LHK6B, no cross-reactivity was observed between these antibodies. The triple staining with RKSE60, LHK6B, and TO-PRO-3 iodide showed subpopulations of K10 expressing cells, K10/K6 co-expressing cells, and K6 only expressing cells. There was a significant difference in the proportion of K6 expression and K10/K6 co-expression between psoriatic and normal skin. Moreover, the proliferative activity of these subpopulations could be quantified by this protocol. We concluded that a triple staining protocol for the assessment of K6, K10, and DNA content, using the monoclonal antibodies LHK6B, RKSE60, and TO-PRO-3 iodide, supplies reliable and reproducible data

Topics: Adult; Aged; Biopsy; Cell Division; DNA; Female; Flow Cytometry; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Humans; Iodine; Keratin-10; Keratins; Male; Middle Aged; Psoriasis; Skin

It has been reported that opioid peptides modulate the differentiation of normal human keratinocytes and that mu-opiate receptors are expressed in human epidermis. The regulation of keratinocyte differentiation is particularly important in psoriasis, and one of the markers for hyperproliferative and differentiating skin diseases is cytokeratin 16. The finding that the endogenous mu-opiate receptor ligand beta-endorphin is increased in serum of patients with psoriasis indicates that the mu-opiate system may play an important role in the pathophysiology of the skin. In this study, we addressed the question whether there is a link between mu-opiate receptor regulation and cytokeratin 16 expression in normal and psoriatic skin. Firstly, we demonstrate that beta-endorphin concentrations between 16 and 1000 nM significantly downregulate mu-opiate receptor expression in epidermis of cultured human skin after 48 h. Secondly, we show that beta-endorphin regulates cytokeratin 16 expression in the epidermis of skin organ cultures exposed to 41-125 nM beta-endorphin for 48 h, leading to elevated cytokeratin 16 production. As expected, the expression of cytokeratin 16 was detected primarily in the suprabasal layer. The same pattern was observed in psoriatic lesional skin, i.e., mu-opiate receptor expression was significantly downregulated and cytokeratin 16 expression upregulated. These results suggest that the mu-opiate receptor system and its ligand beta-endorphin are involved in the pathogenesis of psoriasis, especially in terms of differentiation.

Topics: beta-Endorphin; Down-Regulation; Humans; Immunohistochemistry; Keratins; Organ Culture Techniques; Psoriasis; Receptors, Opioid; Skin; Up-Regulation

Citrulline-containing proteins, mainly originating from keratin K1 and formed by enzymatic deimination of arginine residues, have been identified in the cornified layers of human epidermis. We analyzed the localization and nature of the deiminated proteins in psoriatic epidermis. Immunostaining based on chemical modification of citrulline residues showed that the normal and psoriatic uninvolved epidermis contained deiminated proteins diffusely in the cornified cell layer, whereas the involved epidermis had no detectable or markedly reduced levels of deiminated proteins. Immunolabeling with polyclonal antibodies against a synthetic citrulline-containing peptide corresponding to a deiminated sequence of mouse K1 also suggested markedly decreased deiminated K1 in psoriatic involved lesions. Keratin analyses indicated that deiminated K1 present in normal and psoriatic uninvolved epidermis was not detected in the psoriatic involved epidermis. Double staining with a monoclonal antibody, 34betaB4, and the polyclonal antibodies demonstrated that epidermis with low suprabasal keratin expression was negative for deiminated K1. In contrast, intralesional acrosyringia showing decreased suprabasal keratin immunoreactivity like that of the surrounding psoriatic epidermis showed strong deiminated K1 staining. This suggests that abnormal keratin deimination is restricted to the psoriatic hyperproliferative epidermis, without affecting sweat ductal epithelia.

Topics: Biopsy; Cell Division; Citrulline; Epidermal Cells; Epidermis; Filaggrin Proteins; Humans; Hydrolases; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Protein-Arginine Deiminase Type 4; Protein-Arginine Deiminases; Psoriasis; Skin

Keratin 6 (K6) and keratin 10 (K10) are markers for epidermal hyperproliferation and differentiation, respectively, and are both expressed in the suprabasal layers of the epidermis. They may be co-expressed in different stages of the spreading psoriatic lesion, but single expression can also occur.. To investigate to what extent keratinocytes express K6 and K10, and to what extent they co-express K6 and K10 in different stages of the psoriatic lesion. We studied this in spreading psoriatic plaques.. Three 3-mm punch biopsies were obtained from the inner involved margin of a spreading lesion, from the uninvolved skin immediately adjacent to the spreading plaque, and from the distant uninvolved skin of 8 patients with incipient psoriasis. From 9 healthy volunteers, 3-mm punch biopsies were obtained as controls. After preparation of single cell suspensions of these biopsies, a triple staining protocol was performed with markers for K6 (monoclonal antibody LHK6B), K10 (monoclonal antibody RKSE60) and DNA content (TO-PRO-3 iodide). Subsequently, cells were measured with a flow cytometer and the proportion of the markers was calculated using specific software.. We observed a population of K6/K10-co-expressing cells, but also populations expressing only K6. These subpopulations varied with the involvement of the lesion. There was a statistically significant difference between the inner margin and the outer margin with respect to the proportion of K6- and K10-expressing cells, whereas more K6-positive and K10-negative cells were detected in the inner margin of the lesions. The proportion of K6/K10-co-expressing cells in the inner margin was significantly different from the distant uninvolved skin.. We confirmed that individual keratinocytes in psoriasis can express K6 or K10 depending on their localization in involved or uninvolved skin. There is a unique subpopulation of cells in the psoriatic plaques which co-express K6 and K10. More studies are required to fully understand the pathogenic relevance of co-expression and single expression of K6 and K10.

Topics: Adult; Aged; Cell Division; Female; Flow Cytometry; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Psoriasis; Skin

Topics: Adult; Aged; Antibodies, Monoclonal; Antigens, Nuclear; Biopsy; Cytoskeletal Proteins; DNA-Binding Proteins; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Nuclear Proteins; Psoriasis; Skin

In earlier papers, we suggested that the aggravation of psoriasis by antimalarial drugs (analogous to hypolipidemic drugs) could be initiated by a break in the epidermal barrier. We suggested that these drugs exerted their effect by inhibiting epidermal transglutaminase activity, and supported this hypothesis by demonstrating the effect of hydroxychloroquine sulfate (HCQS) on the morphology of cultured skin and on liver transglutaminase activity. In the present article, we describe, for the first time, the morphologic changes induced by HCQS on cultured skin of psoriatic patients.. Uninvolved (apparently normal) skin explanted from the back of two psoriatic patients was cultured in the presence of 9.2 and 13.8 mM of HCQS for 3 days. The morphologic changes were evaluated in a blind manner. The experiment was repeated twice.. Significant changes in the epidermal morphology of psoriatic skin cultured in the presence of HCQS, compared with skin cultured without the presence of the drug, were observed. The most striking changes were enhanced and irregular keratinization and dermo-epidermal detachment and cleft formation. No parakeratosis or other characteristics of psoriasis were observed.. The first changes caused by HCQS on the cultured skin of psoriatic patients are not characteristic of psoriasis, and include hyperproliferation and enhanced and irregular keratinization. The present experimental study gives further support to the hypothesis that HCQS causes an initial break in the barrier function of the epidermis (probably by inhibiting transglutaminase activity), which is followed by a physiologic response of the epidermis aimed at barrier restoration. This rather nonspecific stimulus to epidermal proliferation is probably sufficient to trigger psoriasis, in vivo, among genetically predisposed patients.

Topics: Antimalarials; Culture Techniques; Dermis; Epidermis; Humans; Hydroxychloroquine; Keratinocytes; Keratins; Melanins; Psoriasis; Skin

We studied the effect of tacalcitol (1 alpha, 24 dihydroxy vitamin D3) ointment on clinical and immunohistochemical efficacy in psoriatic patients during 2 months of treatment. The psoriasis area and severity index decreased significantly after only 1 month and the total body surface index decreased 55% after 2 months. To characterize the epidermal compartment keratin 14, keratin 16, epidermal growth factor receptor, apoptotic and Ki-67 positive cells were examined. After 1 week of treatment no significant changes were found in any of these parameters. After 2 months, keratin 16 reached the levels observed in normal skin and Ki-67 and keratin 14 expression also reduced significantly. Epidermal growth factor receptor staining and the number of apoptotic cells did not alter during treatment. We conclude that tacalcitol is effective in the treatment of plaque psoriasis. Because the main epidermal effect observed immunohistochemically is a reduction in proliferation, a combination therapy using either corticosteroids, vitamin A derivatives or dithranol seems rational.

Topics: Administration, Cutaneous; Adult; Apoptosis; Dermatologic Agents; Dihydroxycholecalciferols; ErbB Receptors; Female; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Keratins; Ki-67 Antigen; Male; Middle Aged; Ointments; Psoriasis; Severity of Illness Index; Treatment Outcome

Several published studies suggest the involvement of immune and inflammatory factors in psoriasis. We recently demonstrated that the number of circulating ICAM-1 + lymphocytes and the levels beta 2-microglobulin are useful parameters in monitoring the activity of the disease. In this study we investigated serum levels of SCCr-Ag in 24 patients with psoriasis in order to determine whether this antigen is a marker of disease activity. Our results demonstrated high serum levels of SCCr-Ag, IL-2, sIL-2R, sCD4, sCD8, sICAM-1 and beta 2-microglobulin in the acute phase of psoriasis. Furthermore, we found a positive correlation of SCC with TBSA, PASI score, sICAM-1 and beta 2-microglobulin. These data demonstrate that serum levels of SCCr-Ag depend on the severity of the disease and correlate with both immunological and inflammatory markers of disease activity. We suggest that expression of SCCr-Ag may be induced by cytokines in the microenvironment of psoriatic lesions, suggesting that SCC-Ag may play a role in the inflammatory process.

Topics: Adult; Antigens, Neoplasm; beta 2-Microglobulin; Biomarkers, Tumor; Body Surface Area; Case-Control Studies; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoradiometric Assay; Intercellular Adhesion Molecule-1; Keratins; Male; Middle Aged; Psoriasis; Serpins; Severity of Illness Index

The mature psoriatic lesion does not necessarily demonstrate changes relevant to early phases of the lesion.. In a model for relapsing psoriasis we examined the epidermal phenotype by means of a panel of immunohistochemical parameters: keratins 14 and 16, epidermal growth factor receptor (EGFR), Ki-67 antigen, and Tdt-mediated Unscheduled Nick End Labeling to detect apoptosis.. In 9 patients, we cleared psoriatic plaques by topical treatment with clobetasol-17-propionate under hydrocolloid occlusion. Relapse (defined as a clinical sum score > or = 6) was awaited. Biopsy specimens of the psoriatic lesion, the cleared skin, the relapsed plaque, and its clinically normal margin were assessed.. Psoriasis recurred after 19+/-6 weeks (mean +/- SEM). During treatment all parameters improved considerably; however, the number of apoptotic cells was not affected. Ki-67 values decreased well below the normal range. At initial relapse, the symptomless skin adjacent to the relapsing lesion (margin) showed a marked expression of keratin 16 and EGFR. Ki-67 expression was increasing in the margin but was below values of the mature lesion. The localization of cycling cells in the first suprabasal layers was a remarkable feature. Keratin 14 expression was increased in the recurrent lesion itself, but not in the symptomless margin.. Keratin 16 and EGFR expression are early phenomena in the evolution of the lesion, and they anticipate epidermal proliferation. The expression of keratin 14 follows overt epidermal hyperproliferation. The present observation in incipient psoriasis lends support to the hypothesis that the basal cell compartment does not have a primary involvement in the initiation of epidermal abnormalities in psoriasis, but that a coordinated sequence of events involving proliferation and differentiation markers in the first suprabasal layers of the epidermis could be the key to the pathogenesis of this puzzling disease.

Topics: Administration, Topical; Adult; Anti-Inflammatory Agents; Apoptosis; Clobetasol; ErbB Receptors; Glucocorticoids; Humans; Immunohistochemistry; In Situ Nick-End Labeling; Keratins; Ki-67 Antigen; Middle Aged; Psoriasis; Recurrence

Skin biopsies from healthy human skin and non-lesional skin from patients with psoriasis were cultured for 24 h and stimulated with interleukin-1 beta (IL-1 beta) and interferon-gamma (IFN-gamma) in a skin organ culture model and the induction of the psoriasiform regenerative epidermal phenotype was analysed using immunostaining. In the presence of IL-1 beta, the psoriasiform regenerative epidermal phenotype was clearly induced. This involved strong up-regulation of the expression of keratin 16, keratin 17, and keratinocyte transglutaminase (TGk) in the suprabasal layers, strong up-regulation and a shift of the expression of keratin 5 and integrin beta 1 from the basal to suprabasal keratinocytes, and induction of the expression of ICAM-1 and HLA-DR on basal keratinocytes. The effects of IL-1 beta in the organ cultures of normal skin could be completely neutralized by anti-IL-1 polyclonal antibodies. The effects of IFN-gamma in healthy and non-lesional psoriatic skin were qualitatively similar to those of IL-1 beta. The IFN-gamma-induced epidermal expression of keratin 17 and TGk could be completely blocked by culturing the biopsies in the presence of IL-1ra or anti-IL-1 antibodies, while the induction of HLA-DR and ICAM-1 was not inhibited. The induction of the psoriasiform regenerative epidermal phenotype by IFN-gamma is partially mediated via endogenous epidermal IL-1.

Topics: Adult; Aged; Epidermis; Female; Humans; Immunoenzyme Techniques; Interferon-gamma; Interleukin-1; Keratinocytes; Keratins; Male; Middle Aged; Organ Culture Techniques; Psoriasis; Recombinant Proteins; Skin; Transglutaminases; Up-Regulation

Psoriasis is a T cell-mediated inflammatory skin disease that has been associated with infections by group A beta-haemolytic streptococci. In a previous study of patients with active psoriasis we demonstrated an increased frequency of circulating Th1-like cells that responded to 20 amino acid (aa) streptococcal M-peptides sharing sequences with human keratin. These cells disappeared after ultraviolet B (UVB)-induced clinical remission. Using T cells from the blood of 17 psoriatic patients and 17 healthy controls we have now compared the numbers of interferon-gamma (IFN-gamma)-producing cells induced by seven 18-20 aa keratin peptides and five corresponding M-peptides. The most frequent and strongest responses were observed to a peptide from keratin 17 that shares ALEEAN sequence with M-protein. The responses to this peptide were stronger than to the corresponding M-peptide containing the ALEEAN sequence. After UVB treatment T cell responses to all the M- and keratin peptides were abolished, while responses to the positive control antigen streptokinase/streptodornase (SK/SD) were not affected. These findings are consistent with the notion that aa sequences which keratin has in common with M-protein may be a major target for autoreactive T cells in psoriasis.

Topics: Amino Acid Sequence; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Carrier Proteins; Cells, Cultured; Epitopes, T-Lymphocyte; Female; Humans; Keratins; Male; Molecular Sequence Data; Peptides; Psoriasis; T-Lymphocytes; Ultraviolet Rays

Epidermolytic hyperkeratosis (EHK) is a genodermatosis caused by mutations in either the keratin 1 (K1) or keratin 10 (K10) genes, and characterized by erythroderma and blistering at birth, with development of a ribbed, ichthyotic hyperkeratosis and palmoplantar keratoderma. A wide variety of mutations within the highly conserved helix termination motifs of the central rod domains of the K1 or K10 genes correlate with the highly variable phenotypic severity observed in EHK. We report a unique EHK-like phenotype exhibiting autosomal dominant inheritance with variable expressivity in four affected individuals in a single family. Clinically, affected individuals manifest transient blistering at birth followed by chronic diffuse palmoplantar keratoderma without transgradiens. Intermittent flares of non-migratory polycylic erythematous psoriasiform plaques which worsen and abate in severity were present in all affected individuals, but showed immense individual variation in both severity and duration, ranging from weeks to months. Histopathologic examination of the psoriasiform plaques demonstrated the characteristic features of EHK. Sequencing of the K1 gene in affected family members revealed a heterozygous A-to-T transversion at nucleotide 1435 within exon 7, converting isoleucine (ATT) to phenylalanine (TTT), (I479F). The mutation resides within the highly conserved helix termination motif of the helix 2B segment of the K1 gene. This unique clinical phenotype and the associated K1 mutation have not been previously described, and it is referred to here as EHK with polycyclic, psoriasiform plaques (EHK/PPP).

Topics: Amino Acid Sequence; Base Sequence; DNA Mutational Analysis; Exons; Female; Genes, Dominant; Humans; Hyperkeratosis, Epidermolytic; Infant, Newborn; Keratins; Male; Mutation; Pedigree; Phenotype; Point Mutation; Psoriasis

Topics: Humans; Keratins; Psoriasis; Retrospective Studies; Streptococcal Infections; T-Lymphocytes; Tonsillectomy; Tonsillitis; Treatment Outcome

Verruciform xanthoma is a rare clinicopathologic entity of uncertain etiology that occurs primarily in the oral mucosa. Aggregates of foam cells in the submucosal stroma or papillary dermis in association with verrucous epithelial hyperplasia are the hallmark of this lesion. Extraoral (cutaneous) occurrence of verruciform xanthoma is much rarer and has been reported mostly in the genital skin. Five cases of extraoral cutaneous verruciform xanthoma (three from the scrotum, one from the penis, and one from the nose) and one histologic "simulant" (from skin of the nose) were studied. The lesions were solitary, raised, or polypoid with cup-shaped craters filled with parakeratotic cells that blended into keratinocytes of an acanthotic and papillomatous epidermis. There was a neutrophilic infiltrate of varying intensity between plump parakeratotic cells and keratinocytes, near the surface of the epidermis. Aggregates of foam cells were present in the papillary dermis, which was highly vascular. A plasma cell predominant infiltrate was seen at the base in a bandlike fashion. Despite the architectural resemblance of verruciform xanthoma to verrucous mucocutaneous lesions related to human papillomavirus infection, it was not detected by either immunohistochemistry, in situ hybridization, polymerase chain reaction, or Southern blot analysis in any case. The foam cells were weakly positive for cytokeratin and for Factor XIIIa but negative for S-100 protein. The KP1 and Mac 387 immunostain showed focal weak staining in foam cells. We postulate that a cascade of events pursue after initial keratinocytic damage attracting neutrophils, with subsequent phagocytosis of necrotic keratinocytic debris by dermal dendrocytes, eventually leading to the ultimate manifestation of the lesion as verruciform xanthoma. The etiologic agent remains elusive, but based on our findings, we conclude that verruciform xanthoma is most likely not a human papillomavirus-associated squamoproliferative lesion and that the foam cells, a histologic hallmark of the lesion, are most likely derived from dermal dendritic cells.

Topics: Adult; Aged; Biomarkers; Female; Genital Diseases, Male; Humans; Immunohistochemistry; In Situ Hybridization; Keratins; Keratoacanthoma; Male; Middle Aged; Nose Diseases; Papillomaviridae; Polymerase Chain Reaction; Psoriasis; Transglutaminases; Xanthomatosis

The aim of this study was to determine if there were characteristic trichogram changes in scalp psoriasis in patients without clinically evident effluvium or alopecia. A total of 45 patients (17 men and 28 women, aged from 15 to 73 years) with clinically and histologically confirmed psoriasis vulgaris with scalp involvement were included. The control group consisted of 60 volunteers (23 males and 37 females aged from 15 to 74 years) with no scalp involvement. Our results from the psoriatic group showed highly increased proportions of dysplastic hair roots. Median proportion was 50% with 95% confidence interval (CI) for median 30-60%, whereas telogen hair ratio was slightly increased-median proportion was 16% with 95% CI for median 15-20%. Within the psoriasis patients' group no statistically significant correlations were found between the proportion of dysplastic hairs and the patients' age, sex, and the intensity and duration of disease. According to the results of this study, the dysplastic hairs in scalp psoriasis are statistically significant much more frequently compared with the control group. Thus, the increased proportion of dysplastic hairs in scalp psoriasis without effluvium or alopecia might be its characteristic trichogram sign.

Topics: Adolescent; Adult; Aged; Female; Humans; Keratins; Male; Middle Aged; Psoriasis; Scalp Dermatoses

Clarification of the pathogenesis of psoriasis requires separate studies of the epidermis, dermis, and inflammatory cells. We previously subcutaneously transplanted a mixture of cultured human keratinocytes and fibroblasts into mice to develop cysts with human skin structures. Using this method, we separately cultured psoriatic and normal keratinocytes and fibroblasts. Four mixtures were prepared: normal keratinocytes and normal fibroblasts (NK/NF); psoriatic keratinocytes and normal fibroblasts (PK/NF); normal keratinocytes and psoriatic fibroblasts (NK/PF); and psoriatic keratinocytes and psoriatic fibroblasts (PK/PF). Each mixture was transplanted into immunodeficient mice to observe formation of cysts and histological changes. The cysts varied in structure depending on the mixture, which suggests that psoriatic keratinocytes and fibroblasts had some abnormalities. Psoriatic fibroblasts may be partially responsible for thickening of the epidermis. Cell differentiation might have been accelerated in psoriatic keratinocytes after transplantation, resulting in the loss of epidermis structures.

Topics: Animals; Cell Communication; Cell Differentiation; Cell Transplantation; Cells, Cultured; Epidermal Cyst; Fibroblasts; Humans; Immunoenzyme Techniques; Keratinocytes; Keratins; Mice; Mice, Inbred BALB C; Mice, Nude; Mice, SCID; Psoriasis; Severe Combined Immunodeficiency; Skin; Transplantation, Heterologous

The biologically active vitamin D analog calcipotriol is effective and safe in the topical treatment of psoriasis, but its exact mechanism of action is unknown.. We investigated expression of 1,25-dihydroxyvitamin D3 receptors, markers for inflammation (CD1a, CD4, CD8, CD11b, CD15; NAP-1/interleukin-8; 55 kd tumor necrosis factor-receptor; intercellular adhesion molecule-1; HLA-DR), proliferation (proliferating cell nuclear antigen, Ki-67), and differentiation (transglutaminase K; involucrin; cytokeratin 16) in psoriatic skin during topical calcipotriol treatment.. For immunohistochemical staining we used the labeled avidin-biotin technique on cryostat-cut sections.. We found a significant increase of 1,25-dihydroxyvitamin D3 receptor expression in epidermal basal keratinocytes of lesional psoriatic skin during calcipotriol treatment. In all patients analyzed, effects on proliferation and differentiation of epidermal keratinocytes were stronger than effects on dermal inflammation. Effects on inflammation were more pronounced in the epidermal than in the dermal compartment.. Our findings indicate that analogs of 1,25-dihydroxyvitamin D3 upregulate their corresponding receptor in human keratinocytes in vivo. This mechanism may be important in the therapeutic efficacy of vitamin D analogs in psoriasis. The differential therapeutic effects in the epidermal and dermal skin compartments may be due to a reduced bioavailability of calcipotriol in the dermal compartment.

Topics: Administration, Cutaneous; Antigens, CD; Antigens, CD1; Biological Availability; Calcitriol; CD11 Antigens; CD4 Antigens; CD8 Antigens; Cell Differentiation; Cell Division; Dermatologic Agents; Epidermis; Gene Expression Regulation; HLA-DR Antigens; Humans; Immunoenzyme Techniques; Immunohistochemistry; Intercellular Adhesion Molecule-1; Interleukin-8; Keratinocytes; Keratins; Lewis X Antigen; Male; Proliferating Cell Nuclear Antigen; Protein Precursors; Psoriasis; Receptors, Calcitriol; Receptors, Interleukin; Receptors, Interleukin-8A; Receptors, Tumor Necrosis Factor; Skin; Transglutaminases; Tumor Necrosis Factor-alpha; Up-Regulation

Desmosomes are predominant among the types of plaque-bearing adhering junctions found in human skin. These structures contain a set of desmosomal cadherins and cytoplasmic plaque proteins, the synthesis of which is differentiation dependent. As plakophilin 1, a member of the armadillo gene family, is an important accessory desmosomal plaque protein, we raised several monoclonal antibodies specific for this protein and applied immunohistochemical and immunoblotting procedures to study the distribution of plakophilin 1 in desmosomes in adult and fetal skin, psoriatic epidermis, various epithelial skin tumors, and keratinocyte sheets grown in culture. In epidermis, the spinous layers were prominently immunostained by plakophilin 1 antibodies, whereas the basal cell layer was only weakly stained and the stratum corneum was entirely unstained. The staining observed in psoriatic epidermis was somewhat heterogeneous. In hair follicles, the outer root sheath (ORS) was delineated in its suprabasal cell layers, with variable staining in its upper and lower parts. All basal cells of the ORS remained unstained, as did upper inner root sheath (IRS) and matrix cells of lower bulb. In eccrine sweat glands, the reaction was confined to inner dermal ductal cells, with the acini remaining unstained. The desmosomal immunostaining observed in basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) was very heterogeneous: In general, junctions in well-differentiated stratified tumor regions were more intensely stained than sections of poorly differentiated and invasively growing BCCs and SCCs. Plakophilin 1 was also prominent in the desmosomes of keratinocyte sheets grown in culture. The cell type-specific, i.e., differentiation-dependent, distribution of desmosomal plakophilin 1 is discussed in relation both to the stratification of the cutaneous epithelia and to tumor differentiation and growth.

Topics: Bowen's Disease; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Desmosomes; Fetus; Foot; Humans; Immunohistochemistry; Keratinocytes; Keratins; Plakophilins; Proteins; Psoriasis; Skin; Skin Neoplasms

Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 are potent drugs for the treatment of psoriasis. It has recently been published that these compounds induce epidermal hyperproliferation in hairless mouse skin.. The aim of our study was to examine the effect of vitamin D3 derivatives on epidermal growth, keratinization and permeability barrier function in vivo.. Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 in isopropanol or in an ointment formula were applied to normal hairless mouse skin. Transepidermal water loss (TEWL), a marker of cutaneous barrier function, and epidermal proliferation were determined at different time points 0-264 h after treatment. In addition, light and electron microscopy studies were performed.. A single treatment in solution led to a transient (2- to 3-fold) increase in TEWL after application of calcipotriol or 1,25(OH)2D3 and to a 3- to 6-fold increase in epidermal proliferation after application of each of the compounds. Repeated applications also resulted in an up to 3-fold increase in TEWL which persisted for 3 days after the end of the treatment. By light microscopy an increase in epidermal thickness was observed. There was no sign of inflammation. Electron microscopy studies showed the formation of a transitional cell zone as a sign of a premature keratinization.. These results demonstrate that in normal mouse skin vitamin D3 and its analogues disrupt the epidermal permeability barrier by induction of epidermal proliferation and premature keratinization but without morphological signs of inflammation.

Topics: 1-Propanol; Administration, Cutaneous; Animals; Calcitriol; Cell Division; Cholecalciferol; Dermatologic Agents; Dihydroxycholecalciferols; Epidermis; Keratins; Male; Mice; Mice, Hairless; Microscopy, Electron; Ointments; Permeability; Psoriasis; Skin; Time Factors; Water Loss, Insensible

The peripheral changes in the uninvolved skin adjacent to the extending psoriatic lesions may represent early events. The sequence of these events remains controversial. In the present study we evaluated epidermal and dermal aspects of the margin of the progressive psoriatic plaque, the distant uninvolved skin and normal healthy skin, using immunohistochemistry with markers for keratinization, proliferation and connective tissue: filaggrin, involucrin, Ki-67 and tenascin. The results showed that: (i) processes in distant uninvolved skin were comparable with the observations in normal skin; (ii) in the margin zone of the spreading psoriatic lesion, following the increased appearance of tenascin, the transition into parakeratosis, abnormal expression of filaggrin, involucrin and recruitment of cycling epidermal cells, happened simultaneously. The simultaneous normalization of these epidermal processes might be a consequence of a signal which is simultaneously transduced to the basal and suprabasal cell layers of the epidermis. Based on the present results and earlier findings, we would like to propose a triple stage model for the development of the psoriatic lesion: Stage 1, involvement of the stroma; stage II, inflammatory infiltrate formation and penetration into the upper layers of the epidermis, with suprabasal expression of keratin 16; stage III, recruitment of cycling epidermal cells and abnormal terminal differentiation. Further studies are required on the regulation of tenascin expression, focusing on factors derived from the stroma affecting both recruitment of cycling epidermal cells, involucrin and filaggrin expression. An intermediate step in the dermo-epithelial interrelation is the inflammatory infiltrate, penetrating into the most superficial zone of the epidermis, and the suprabasal expression of keratin 16.

Topics: Adult; Aged; Biopsy; Cell Division; Epidermal Cells; Epidermis; Female; Filaggrin Proteins; Gene Expression Regulation; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Ki-67 Antigen; Male; Middle Aged; Protein Precursors; Psoriasis; Skin; Tenascin

Psoriasis is a T-cell mediated inflammatory skin disease which has been associated with group A, beta-haemolytic streptococcal infections. Four 20 a.a. long M6-peptides sharing 5-6 a.a. sequences with human epidermal keratins were identified. To investigate the role of potentially cross-reactive T cells in the pathogenesis of psoriasis, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) responses of circulating T cells to these peptides were analysed by ELISPOT and RT-PCR in 14 psoriatic patients, 12 healthy individuals and six patients with atopic dermatitis (AD). Untreated psoriatic patients' responses were significantly higher to these peptides than healthy and AD controls, while responses to a control M6-peptide, not sharing sequences with keratin, were negligible in all groups. No difference was found in response to streptokinase/streptodornase (SK/SD). M6-protein and peptides exclusively elicited IFN-gamma production, with little IL-4 production, even in AD patients. Interferon-gamma responses to all the M6-peptides were abolished after successful treatment of psoriatic patients, but responses to SK/SD were unaffected. The results indicate that active psoriasis is associated with Th1-like cells responding to streptococcal M6-peptides sharing sequences with human epidermal keratin. This is consistent with the hypothesis that psoriasis may be induced and exacerbated in susceptible individuals by M-protein specific Th1-like cells that cross-react with human epidermal keratin.

Topics: Adult; Amino Acid Sequence; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Carrier Proteins; Epidermis; Female; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-4; Keratins; Lymphocyte Activation; Male; Molecular Sequence Data; Psoriasis; Streptococcus pyogenes; Streptodornase and Streptokinase; T-Lymphocytes

The potent calciotropic hormone calcitriol (1,25-dihydroxyvitamin D3, 1,25(OH)2D3) has been shown to be very effective and safe in the topical treatment of psoriasis. In vitro, 1,25(OH)2D3 inhibits proliferation and stimulates differentiation of human keratinocytes. Increasing evidence suggests an immunoregulatory function of this potent steroid hormone. To further characterize the biological effects of topical calcitriol treatment in psoriasis, we have analyzed immunohistochemically the expression of markers for epidermal proliferation (proliferating cell nuclear antigen=PCNA) and differentiation (transglutaminase K, involucrin, cytokeratin 16), as well as inflammation (CD1a, 55 kDa TNF-receptor, NAP-1/IL-8) in calcitriol-treated psoriatic skin in situ. Our findings strongly support the hypothesis that calcitriol modulates keratinocyte proliferation/differentiation as well as inflammation in human skin in vivo. The immunoreactivity of markers for epidermal proliferation and differentiation, as well as of CD1a and NAP-1/IL-8, changed after 8 weeks of calcitriol treatment almost completely to the pattern characteristic for non-lesional psoriatic skin, while a large number of 55 kDa TNF-receptor positive cells could be found in the dermal compartment.

Topics: Administration, Topical; Antigens, CD1; Calcitriol; Cell Differentiation; Cell Division; Humans; Immunohistochemistry; Interleukin-8; Keratinocytes; Keratins; Proliferating Cell Nuclear Antigen; Protein Precursors; Psoriasis; Receptors, Tumor Necrosis Factor; Skin; Transglutaminases

Amphiregulin (AR) is a heparin-binding, heparin-inhibited member of the epidermal growth factor (EGF) family and an autocrine growth factor for human keratinocytes. Previous studies have shown that AR expression is increased in psoriatic epidermis. To test the hypothesis that aberrant AR expression is central to the development of psoriatic lesions, we constructed a transgene (K14-ARGE) encoding a human keratin 14 promoter-driven AR gene. Our results indicate that transgene integration and subsequent expression of AR in basal keratinocytes correlated with a psoriasis-like skin phenotype. Afflicted mice demonstrated shortened life spans, prominent scaling and erythematous skin with alopecia, and occasional papillomatous epidermal growths. Histologic examination revealed extensive areas of marked hyperkeratosis with focal parakeratosis, acanthosis, dermal and epidermal lymphocytic and neutrophilic infiltration, and dilated blood vessels within the papillary dermis. Our results reveal that AR exerts activity in the skin that is distinct from that of transgenic transforming growth factor-alpha or other cytokines, and induces skin pathology with striking similarities to psoriasis. Our observations also link the keratinocyte EGF receptor-ligand system to psoriatic inflammation, and suggest that aberrant expression of AR in the epidermis may represent a critical step in the development or propagation of psoriatic lesions.

Topics: Amphiregulin; Animals; CD3 Complex; EGF Family of Proteins; Epidermis; Gene Expression Regulation; Glycoproteins; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Keratins; Ki-67 Antigen; Mice; Mice, Transgenic; Psoriasis

Lesional psoriatic epidermis displays a number of phenotypic changes that are distinct from the differentiation program found in normal interfollicular epidermis. In psoriatic epidermis, keratinocytes are hyperproliferative and several differentiation-associated molecules are expressed that are absent in normal skin (e.g., cytokeratins (CK) 6, 16, and 17, and the epidermal proteinase inhibitor SKALP/ elafin). In addition, several molecules which are normally restricted to the stratum granulosum are strongly upregulated in the stratum spinosum (e.g., psoriasis-associated fatty acid binding protein (PA-FABP), psoriasin, involucrin, and transglutaminase). The aim of this study was to develop in vitro culture systems which (a) would allow to study the induction of normal and psoriatic differentiation pathways, and (b) would be amenable for screening of antipsoriatic drugs. Here we have investigated several models for induction of differentiation with respect to the expression of markers for the normal and psoriatic phenotype. Cell cycle parameters and expression levels of CK1, CK10, CK16, SKALP/elafin, transglutaminase, involucrin, psoriasin, and PA-FABP were assessed in these models using flow cytometry, immunocytochemistry, and Northern blot analysis. We observed that induction of differentiation with fetal calf serum resembled the psoriatic phenotype (sustained hyperproliferation; high levels of CK16, SKALP/elafin, transglutaminase, and involucrin; moderate psoriasin expression), whereas differentiation induced by growth factor depletion in a confluent culture resembled the normal differentiation phenotype (low proliferative rate; high expression levels of CK1 and CK10; moderate expression of involucrin and transglutaminase; low expression levels of SKALP/elafin and CK16; absence of psoriasin). We propose that these models can be used to study expression and pharmacological modulation of selected differentiation genes and the coordinated expression of sets of genes associated with epidermal differentiation programs.

Topics: Blotting, Northern; Carrier Proteins; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Culture Media; Fatty Acid-Binding Protein 7; Fatty Acid-Binding Proteins; Flow Cytometry; Humans; Immunohistochemistry; Keratinocytes; Keratins; Membrane Proteins; Myelin P2 Protein; Neoplasm Proteins; Phenotype; Protein Precursors; Proteinase Inhibitory Proteins, Secretory; Proteins; Psoriasis; RNA, Messenger; Serine Proteinase Inhibitors; Transglutaminases; Tumor Suppressor Proteins

The expression of SPRR (small proline-rich protein) was investigated in normal human skin and in diseased skin from patients with psoriasis, squamous cell carcinoma, basal cell epithelioma, naevus pigmentosus, ichthyosis vulgaris and several inflammatory skin diseases, by immunohistochemical staining. A polyclonal antibody was raised against a synthetic peptide for a C-terminal common region for SPRR1 and SPRR3. In immunoblot analysis, a positive band of 18 kDa was detected, which showed the presence of SPRR1 in human epidermal keratinocytes. In normal epidermis, positive staining for SPRR was observed in keratinocytes in the granular layer and the uppermost or two spinous cell layers, with no staining of the other spinous or basal layers. The staining was obvious at the cell periphery, weak at the cytoplasm, and absent in the nucleus. Staining was observed in several outer layers of the follicular infundibulum to the isthmus. No staining was detected in the inner root sheath of the hair follicles, hair matrix, sebaceous gland, eccrine gland, eccrine duct, melanocytes, Langerhans cells or fibroblasts. The arrectores pilorum, striated muscles, muscle layers of vessels, and myoepithelia of eccrine gland, were weakly stained. In psoriatic skin, stained keratinocytes were distributed in the spinous cell layers except for the basal layer. In ichthyosis vulgaris, SPRR was barely expressed in the uppermost living cell layers of the epidermis. In epidermolytic hyperkeratosis, degenerated squamous cells widely expressed SPRR. In Darier's disease, dyskeratotic cells were clearly stained. In squamous cell carcinoma, staining was observed in keratotic cells around horny pearls. In basal cell epithelioma, naevus pigmentosus, and malignant melanoma, the tumour cells or naevus cells were not stained. The distribution of SPRR was similar to that of involucrin in normal and several diseased skin, except for ichthyosis vulgaris. We conclude that SPRR is expressed in close association with epidermal differentiation in normal skin and skin diseases. The alteration of the expression of the proteins correlated to terminal differentiation, and differs from disease to disease.

Topics: Adult; Aged; Amino Acid Sequence; Cell Differentiation; Cornified Envelope Proline-Rich Proteins; Dermatitis; Epidermis; Female; Humans; Immunoblotting; Immunoenzyme Techniques; Keratins; Male; Membrane Proteins; Middle Aged; Molecular Sequence Data; Proteins; Psoriasis; Skin; Skin Diseases; Skin Neoplasms

Keratin K17, the myoepithelial keratin, is expressed in psoriasis but is not present in healthy skin. Psoriasis is associated with production of gamma interferon (IFN gamma), which induces the expression of keratin K17 by activating transcription factor STAT1. Our hypothesis states that the induction of K17 is specific for the inflammatory reactions associated with high levels of IFN gamma and activation of STAT1. One of the corollaries of the hypothesis is that the STAT1-activating cytokines should induce the expression of keratin K17, whereas those cytokines that work through other mechanisms should not. Furthermore, because the STAT activation pathway is dependent upon protein phosphorylation events, phosphorylation inhibitors should attenuate the induction of keratin K17, whereas protein phosphatase inhibitors should augment it. To test this hypothesis, we analyzed lesional samples of inflammatory diseases using immunofluorescence, transfected keratinocytes with K17 gene promoter DNAs in the presence of various cytokines, and followed nuclear translocation of STAT1 in keratinocytes using specific antibodies. Confirming the hypothesis, we found that K17 is induced in psoriasis and dermatitis caused by delayed type hypersensitivity, which are associated with high levels of IFN gamma, but not in samples of atopic dermatitis, which is not. Two cytokines, interleukin-6 and leukemia inhibitory factor, which can induce phosphorylation of STAT1, can also induce K17 expression, whereas interleukin-3, interleukin-4, interleukin-10, and granulocyte macrophage colony stimulating factor have no effect on K17 expression. As expected, staurosporine and genistein inhibited, whereas okadaic acid augmented, the induction of K17 by IFN gamma. Our data indicate that in inflammatory skin diseases, lymphocytes, through the cytokines they produce, differently regulate not only each other, but also keratin gene expression in epidermis one of their target tissues.

Topics: Cytokines; Dermatitis; Dermatitis, Atopic; DNA-Binding Proteins; Enzyme Inhibitors; Epidermis; Growth Inhibitors; Humans; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-6; Keratins; Leukemia Inhibitory Factor; Lymphokines; Phosphoprotein Phosphatases; Promoter Regions, Genetic; Protein Kinase Inhibitors; Psoriasis; STAT1 Transcription Factor; Trans-Activators; Transcription, Genetic

Renewal of epidermal cells is a highly coordinated process in which terminal differentiation balances the proliferative rate in the germinative compartment. Exact quantitative data on cell cycle parameters of normal human epidermis are fragmentary, and do not allow firm conclusions on issues such as cell cycle time, duration of the various cell cycle phases, and the pool sizes of the different cellular populations. As part of a study on bone marrow cell cycle kinetics, 14 lymphoma patients were infused with the thymidine analogue iododeoxyuridine (IdUrd). This provided us with the unique opportunity to study the cell cycle kinetics in normal epidermis obtained from these patients. Single epidermal cell suspensions were prepared from skin, stained with propidium iodide (PI) for relative DNA content, and simultaneously labeled with an anti-IdUrd antibody to detect DNA-synthesizing cells. In parallel samples suprabasal cells were analyzed by labeling with an anti-cytokeratin 10 antibody. Analysis was performed using bivariate flow cytometry. The results showed that 3.5% of the total epidermal cell population was in S-phase. An S-phase duration of 9.7 +/- 0.6 h and a cytokeratin 10-positive pool size of 59.6 +/- 4.6% were obtained. The duration of the G1-phase and the G2M-phase were calculated to be 7.6 +/- 2.0 h and 11.1 +/- 2.0 h, respectively. From these data a total cell cycle time can be calculated of 28.4 h. Combining this data with previous findings we were able to determine a similar cell cycle time of 27.8 h, and pool sizes of the epidermal cells: 30% quiescent (resting, G0) and 10% cycling cells. The implications of these findings for the interpretation of deviations in growth control as found in hyperproliferative skin diseases (e.g. psoriasis) are discussed.

Topics: Antibodies, Monoclonal; Cell Cycle; DNA; Flow Cytometry; Humans; Idoxuridine; Keratinocytes; Keratins; Kinetics; Propidium; Psoriasis; S Phase; Skin

In order to confirm and further explore the significance of the overexpression of the CRABP II (cellular retinoic acid binding protein type II) and psoriasin genes in psoriatic versus normal skin, we examined the mRNA expression levels of these two genes by in situ hybridization in skin samples from psoriatic plaques and in one case from the border between a psoriatic plaque and uninvolved skin. Both genes were markedly upregulated in lesional skin, with a shift from low to high expression in the transitional zone of the plaque. Expression of the cytokeratin 1 (K1) gene was, in contrast, high in normal skin and decreased in the transition from uninvolved skin to psoriatic plaque, Examination of mRNA levels of CRABP II and psoriasin in other hyperproliferative and inflammatory skin diseases showed high expression of psoriasin, and in some cases also of CRABP II, in atopic dermatitis, mycosis fungoides, Darier's disease and inflammatory lichen sclerosus et atrophicus. In atrophic lesions of lichen sclerosus et atrophicus that lacked an inflammatory infiltrate, these changes were only weakly expressed. These findings demonstrate that altered epidermal gene expression of K1, psoriasin and CRABP II is not disease-specific and may reflect instead an altered state of epidermal differentiation and/or may be linked to the inflammation and cellular infiltration common to all the conditions studied.

Topics: Biopsy; Calcium-Binding Proteins; Case-Control Studies; Humans; Keratins; Psoriasis; Receptors, Retinoic Acid; RNA, Messenger; S100 Calcium Binding Protein A7; S100 Proteins

Bone morphogenetic protein-6 (BMP-6) belongs to the family of TGF-beta-related growth factors. In the developing epidermis, expression of BMP-6 coincides with the onset of stratification. Expression persists perinatally but declines after day 6 postpartum, although it can still be detected in adult skin by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. We constitutively overexpressed BMP-6 in suprabasal layers of interfollicular epidermis in transgenic mice using a keratin 10 promoter. All mice expressing the transgene developed abnormalities in the skin, indicating an active transgene-derived factor. Depending on the pattern of transgene expression, the effects on proliferation and differentiation were completely opposite. Strong and uniform expression of the BMP-6 transgene resulted in severe repression of cell proliferation in embryonic and perinatal epidermis but had marginal effects on differentiation. Weaker and patchy expression of the transgene evoked strong hyperproliferation and parakeratosis in adult epidermis and severe perturbations of the usual pattern of differentiation. These perturbations included changes in the expression of keratins and integrins. Together with an inflammatory infiltrate both in the dermis and in the epidermis, these aspects present all typical histological and biochemical hallmarks of a human skin disease: psoriasis.

Topics: Acanthosis Nigricans; Animals; Animals, Newborn; Antigens, CD; Bone Morphogenetic Protein 6; Bone Morphogenetic Proteins; Cell Differentiation; Cell Division; Epidermal Cells; Epidermis; Gene Expression Regulation, Developmental; Humans; Integrin alpha6; Keratin-10; Keratins; Keratosis; Mice; Mice, Transgenic; Mouth Mucosa; Promoter Regions, Genetic; Psoriasis; RNA, Messenger; Skin; Stomach

Psoriasis vulgaris is associated with the HLA-Cw6 and Cw7 antigens. However, it has not yet been clarified if the HLA-Cw6 and Cw7 genes themselves are the susceptible gene related to this disease or if it is some other non-HLA gene in a linkage disequilibrium with these HLA-C alleles. The S gene, recently identified in the HLA class I region 160 kb telomeric of HLA-C, encodes a keratin-like protein and is expressed specifically in the granular layer of the epidermis. Therefore, it is tempting to speculate that the S gene is one of the strong candidate genes responsible for the pathogenesis of psoriasis vulgaris. Direct sequencing of the first and second exon of the S gene after polymerase chain reaction (PCR) amplification has allowed the identification of two diallelic polymorphic sites in exon I and seven diallelic polymorphic sites in exon 2, three among which result in amino acid exchanges, a Ser-Phe substitution at amino acid position 186, a Gly-Val substitution at position 393 and a Ser-Leu substitution at position 394. No significant difference in the dimorphic distributions of the S gene was observed between the patients with psoriasis vulgaris and healthy controls, suggesting that the susceptible gene for psoriasis is not the S gene itself.

Topics: Disease Susceptibility; Genes, MHC Class I; Genetic Markers; HLA-C Antigens; Humans; Keratins; Polymorphism, Single-Stranded Conformational; Psoriasis

Psoriasis is characterized by immune activation, increased proliferation and abnormal differentiation of keratinocytes. The reported anti-psoriatic mechanisms of action in vivo of vitamin D analogues include reduction of keratinocyte proliferation and induction of keratinocyte terminal differentiation. We investigated whether the anti-psoriatic effect of the natural active vatamin D analogue, calcitriol, applied topically, is due to direct effects on keratinocytes alone or also due to immunoregulatory effects of calcitriol. Psoriasis patients were treated with topical calcitriol (0.005%) and a vehicle control for 8 weeks. Disease activity was assessed by a severity index and quantitative histopathological markers. In vitro studies of lymphocyte proliferation and gamma interferon secretion and of keratinocyte proliferation complemented the clinicohistopathologic studies. A heterogeneous response to calcitriol treatment could be segregated based upon elimination of K-16 keratin expression. Calcitriol treatment decreased keratinocyte proliferation, normalized keratinocyte differentiation and decreased immune activation in plaques. The histologic response to vitamin D treatment of psoriasis includes suppression of both immune and keratinocyte activation in situ. These studies provide a basis for rational combination of anti-psoriatic treatments and for the design of new vitamin D analogues to treat psoriasis.

Topics: Administration, Topical; Calcitriol; Cell Differentiation; Chemotaxis, Leukocyte; Epithelium; Humans; Inflammation; Keratinocytes; Keratins; Psoriasis; T-Lymphocytes

Psoriatic hyperproliferative epidermis is characterized by a regular elongation of rete ridges, accompanied by altered keratinization. Another notable finding is close positioning of the vasculature to the suprapapillary epidermis. These architectural/morphological changes are naturally described by a concept of epidermal remodelling based on decreased epidermal turnover time. The recently described positioning of stem cells to the tips of dermal papillae fits nicely with this concept.

Topics: Cell Division; Epidermis; Humans; Keratinocytes; Keratins; Models, Biological; Psoriasis; Regional Blood Flow; Stem Cells

The environmental contaminant dioxin exerts most of its effects by activating the aryl hydrocarbon receptor (AhR). The AhR is considered to play not only a role in the regulation of xenobiotic metabolism, but also for development, growth, and differentiation. The transcript levels of the AhR and its associated translocator protein (ARNT) were found to increase with ongoing differentiation in the human keratinocyte cell line HaCaT. Correspondingly, in situ hybridization studies in normal human skin revealed an absence of AhR-expression in proliferating basal cells and increasing transcript levels in upper cell layers, in dependence of keratinocyte differentiation. AhR expression in differentiation-deficient hyperproliferative psoriatic skin was markedly decreased. When keratinocytes were continuously treated with 1 microM retinoic acid (RA), the upregulation of AhR- and ARNT-mRNA levels was inhibited as was keratin 4-expression, a marker of HaCaT-keratinocyte differentiation. In contrast, treatment of already differentiated cells with RA did not down-regulate these transcript levels. The mRNA levels of the prevalent retinoic acid receptors in keratinocytes, RAR gamma and RXR alpha, were not influenced by the process of differentiation or by addition of RA. Our data suggest that the regulation of AhR-, ARNT- and keratin 4-expression by RA is indirect and mediated by a yet to be identified factor.

Topics: Aryl Hydrocarbon Receptor Nuclear Translocator; Cell Differentiation; Cell Division; Cells, Cultured; DNA-Binding Proteins; Gene Expression Regulation, Developmental; Humans; Keratinocytes; Keratins; Polychlorinated Dibenzodioxins; Psoriasis; Receptors, Aryl Hydrocarbon; Receptors, Retinoic Acid; Retinoic Acid Receptor gamma; Retinoid X Receptors; RNA, Messenger; Time Factors; Transcription Factors; Tretinoin

The immortalized human keratinocyte cell line HaCaT was used to assess the effect of interferon-gamma (IFN-gamma) on expression of keratin K17. Both IFN-gamma and K17 have been implicated in the pathophysiology of psoriasis. Western and quantitative enzyme-linked immunosorbent assay analyses demonstrated increasing induction of K17 protein by 48 h exposure to IFN-gamma at concentrations of 10, 50, and 250 U/ml. At 50 U/ml IFN-gamma, immunohistochemical analysis revealed numerous K17-positive foci, whereas in situ hybridization demonstrated K17 message in the majority of cells. In addition, at low (5 U/ml) concentrations of IFN-gamma, cell proliferation and protein synthesis decreased, as determined by 3H-thymidine labeling and 14C-amino acid uptake. These data suggest that aberrant K17 expression observed in psoriatic lesions may be a consequence of IFN-gamma overexpression, and that the HaCaT cell line may be a useful in vitro model system to elucidate the underlying mechanisms.

Topics: Antigens, Differentiation; Blotting, Western; Cell Line; Enzyme-Linked Immunosorbent Assay; Humans; Interferon-gamma; Keratinocytes; Keratins; Psoriasis; RNA, Messenger; Up-Regulation

The acid cytokeratin K17 is inducible by interferon-gamma (IFN-gamma), a characteristic unique for cytokeratins analysed so far. In this report, we analysed the molecular basis of K17 expression by IFN-gamma in epithelial cells. The 5'-flanking region of the K17 gene (positions -1762 to -13), cloned in front of a chloramphenicol acetyl transferase (CAT) reporter gene construct, conferred responsiveness to IFN-gamma but not IFN-alpha in transient transfection assays. Sequence analysis revealed three putative gamma-interferon activation sites (GAS). Band-shift assays and transient transfections with CAT reporter gene constructs were used to characterize and to dissect the functional importance of each of the putative GAS elements. In the band shift assay, GAS3 (positions -1528 to -1515) was found to bind GAF/STAT91 and to compete with tryptophanyl-tRNA synthetase (IFP53/WRS)-GAS for binding to GAF; in contrast, GAS1 (positions -183 to -171) and GAS2 (positions -290 to -277) were neither able to bind to nor to compete for GAF/STAT91. However, deletion constructs and mutational analysis of CAT reporter gene constructs harbouring the 5'-flanking region (positions -1762 to -111) in front of the heterologous promoter revealed that the distal GAS3 site was dispensible, but that alteration of the GAS1 element rendered the promoter uninducible by IFN-gamma. Surprisingly, transfection of a CAT-reporter gene construct harbouring a promoter segment (positions -111 to +13) devoid of the GAS elements revealed enhanced CAT-gene expression upon IFN-gamma treatment. The interaction of GAS1 with the interferon-responsive promoter region in the physiological context remains to be clarified.

Topics: Base Sequence; Cell Line; Chloramphenicol O-Acetyltransferase; DNA Mutational Analysis; Enhancer Elements, Genetic; Gene Expression Regulation, Enzymologic; HeLa Cells; Humans; Interferon-gamma; Keratins; Molecular Sequence Data; Psoriasis; Sequence Deletion; Transcriptional Activation

Specimens of trichilemmal cyst, malignant trichilemmoma, keratoacanthoma, and epidermal cyst were examined to characterize keratin peptides in hair-follicle-derived tumors. Keratins were extracted from the specimens and analyzed by two-dimensional gel electrophoresis and densitometry; the results were then compared with those for normal epidermis, the outer root sheath of hair follicles, psoriatic epidermis, and various nonfollicular cutaneous epithelial tumors. The specific nonfollicular tumors examined were squamous cell carcinoma, Bowen disease, actinic keratosis, eccrine porocarcinoma, and sebaceous carcinoma. Immunohistochemistry also was performed with a few anti-keratin monoclonal antibodies. As a general rule, K6 and K16 were expressed in hyperproliferative conditions, such as epidermal tumors, and K17 was coexpressed in the same lesions. The ratio of K16 to K17 in many epithelial skin tumors has been unclear until now. K17 content exceeded K16 content in most follicular tumors, whereas in almost all the nonfollicular tumors and the psoriatic epidermis, K17 levels were less than or about equal to K16 levels. There was a significant difference in the ratio of K16 to K17 between follicular and nonfollicular skin tumors. These results indicate that alterations in the content of these keratins may be associated with follicular differentiation.

Topics: Epidermis; Hair; Humans; Immunohistochemistry; Keratins; Keratoacanthoma; Neoplasms, Basal Cell; Psoriasis; Skin Neoplasms

Topics: Alleles; Epidermis; Genes, MHC Class I; HLA-C Antigens; Humans; Keratinocytes; Keratins; Membrane Glycoproteins; Psoriasis; Serine; Skin Diseases; Vitronectin

During the epidermis maturation different keratin genes are expressed in strictly definite sequences according to the level of epidermis. In psoriasis this sequence in deranged (permutation) Perhaps this is due to implication of homeobox system similar to what occurs in (Ftz) gene. This problem is reviewed and discussed in this paper. All keratin as well as involukrine genes have been sequenced. Regions of K5 gene have been found which play regulatory role in transcription. It has been achieved using transfection K5 into keratinocytes. Psoriatic injury reversed by yEPP peptide of chick Y. sack cells. It is well established that protooncogenes are involved in transcription regulation. Expression of c-myc is increased in psoriatic plagues whereas that of c-fos and c-jun is decreased. This fact may be regarded as an indirect evidence for abnormal functioning c-myc in regulation. It is not ruled out that the system implicated in shedding in reptiles recapitulate in psoriasis.

Topics: Adolescent; Adult; Aged; Epidermis; Female; Gene Expression Regulation; Genes, fos; Genes, Homeobox; Genes, jun; Genes, myc; Humans; Keratins; Male; Middle Aged; Psoriasis; Transcription, Genetic

Psoriasis is characterized by immune activation and increased epidermal proliferation. Cyclosporine acts by reducing T lymphocyte numbers and lymphokine production. Anthralin inhibits keratinocyte proliferation.. We investigated whether topical anthralin would augment clearing of psoriasis produced by systemic cyclosporine.. Twelve patients with psoriasis were treated with cyclosporine (5 mg/kg per day). Patients applied anthralin only to plaques on half of their body. They were treated until a remission or maximum benefit was achieved. Disease activity was assessed by a severity index and quantitative histopathologic markers.. Of the 12 patients, the skin of five cleared within 10 weeks irrespective of anthralin use. The other seven (slow responders) continued treatment for a mean of 18 weeks. Slow responders had a significantly lower severity index, a thinner epidermis, fewer CD8+ cells, and fewer proliferating keratinocytes on the anthralin-treated side than on the non-anthralin-treated side.. The combination of cyclosporine and topical anthralin is effective in patients who are slow to respond to cyclosporine alone.

Topics: Administration, Cutaneous; Administration, Oral; Adult; Anthralin; CD8-Positive T-Lymphocytes; Cell Division; Cyclosporine; Drug Administration Schedule; Drug Therapy, Combination; Epidermis; Female; Humans; Keratinocytes; Keratins; Lymphocyte Count; Lymphokines; Male; Middle Aged; Psoriasis; Remission Induction; Skin; T-Lymphocytes

Increasing evidence suggests that the retinoid-X receptors RXR(-alpha,-beta,-gamma) play a crucial part in regulating the transcriptional activity of several steroid hormone receptors, including 1,25-dihydroxyvitamin D3 receptors (VDR) and retinoic acid receptors (RAR-alpha,-beta,-gamma). We developed a new technique for immunohistochemical in situ detection of RXR receptors, and investigated the localization of RXR-alpha in normal and psoriatic human skin using a recently raised corresponding specific antibody. RXR-alpha positive cells related to the skin were phenotyped by sequential sections and a double-labelling procedure for the simultaneous demonstration of this nuclear receptor and cell membrane antigens, as well as cytokeratin 10, HLA-DR and vimentin. Our findings indicate that: (i) RXR-alpha is strongly expressed in normal and psoriatic human skin; (ii) most of the cell types in normal human skin, including keratinocytes, melanocytes, fibroblasts and skin immune cells such as Langerhans cells, reveal strong nuclear immunoreactivity for RXR-alpha, with less cytoplasmic staining; (iii) altered levels or distribution of RXR-alpha in the skin do not appear to be involved in the genesis of psoriasis vulgaris, but subepidermal and subcellular distribution suggest a function of RXR-alpha in the transition from proliferation to differentiation in epidermal keratinocytes; (iv) expression in the hair follicle points to a contribution from RXR-alpha to hair growth.

Topics: Adult; Animals; Epidermis; HLA-DR Antigens; Humans; Immunoenzyme Techniques; Keratinocytes; Keratins; Male; Mice; Nuclear Proteins; Psoriasis; Receptors, Retinoic Acid; Retinoid X Receptors; Sebaceous Glands; Skin; Transcription Factors; Vimentin

Keratinocyte differentiation in psoriasis was examined using a panel of monospecific monoclonal antibodies to keratins (K), including two recently developed monoclonal antibodies raised to carboxy terminal peptides of K6 (LL020) and K16 (LL025). Keratinocytes from normal skin, untreated psoriatic plaques and non-lesional psoriatic skin, were cultured using multiple in vitro systems. Time-lapse cinephotography was used to measure the intermitotic time of normal and psoriatic keratinocytes in both low calcium-defined and serum-containing media. The intermitotic time did not differ significantly between psoriatic and normal keratinocytes. Keratin expression of psoriatic and normal keratinocytes in vitro was examined by both gel electrophoresis and immunocytochemistry. K6, K16 and K17 were detected suprabasally in all culture systems in vitro, but only in interfollicular psoriatic epidermis in vivo, and not in normal skin. Small subpopulations of keratinocytes expressed simple epithelial keratins K7, K8, K18 and K19 in cultures on plastic substrates, but these keratins were absent in skin equivalents of normal or psoriatic skin. No psoriasis-specific pattern of differentiation was found in vitro. As the K6 peptide antibody reacted with basal cells of normal skin, probably due to K5 cross-reactivity, K16 expression determined by LL025 was found to be the most sensitive indicator of the psoriatic state of differentiation, and this antibody is recommended for future work on psoriasis. K17 had a distinct pattern of tissue distribution in normal skin: K17, but not K16, was present in basal myoepithelial cells in sweat glands, and the deep outer root sheath, but K17 distribution paralleled that of K16 in suprabasal psoriatic epidermis. As keratins K6, K16 and K17 are expressed in keratinocyte hyperproliferation, when high levels of certain cytokines are also expressed, the role of growth factors and regulatory nuclear transcription factors in the control of K6, K16 and K17 expression in psoriasis requires further study, in order to provide insight into the relationship between proliferation and differentiation.

Topics: Biomarkers; Cell Differentiation; Cell Division; Cells, Cultured; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Humans; Immunoenzyme Techniques; Keratinocytes; Keratins; Photography; Psoriasis; Skin

The concept of epidermal architecture that depends on epidermal turnover time is described. The epidermal architecture is determined, not by a simple proliferative condition, but rather by an epidermal turnover time (that depends both on cell number as well as on the proliferative condition). This is in relation to 'keratinization process' that requires a definite time to be completed. Using the concept, hyperproliferative 'psoriasiform' epidermis and hypoproliferative 'atrophic' epidermis are naturally described.

Topics: Atrophy; Cell Count; Cell Division; Epidermal Cells; Epidermis; Hypertrophy; Keratins; Models, Biological; Psoriasis; Time Factors

Increased levels of IgA antibodies to cytokeratin-18 (CK-18) and epidermal keratins (EpK) in the sera of patients with rheumatoid arthritis (RA) have been demonstrated previously. In the present study investigations were carried out to determine whether levels of these autoantibodies were also raised in the spondyloarthropathies, and whether there was any association with particular disease manifestations.. Using specific enzyme linked immunosorbent assays (ELISA) measurements were taken of IgA, IgG and IgM antibodies to EpK and to CK-18 in the sera of patients with psoriatic arthropathy, ankylosing spondylitis (AS), Reiter's syndrome, psoriasis and in normal subjects.. IgA antibodies to both EpK and CK-18 were significantly increased in sera from patients with psoriasis and psoriatic arthropathy but not in the sera from the patients with AS or Reiter's syndrome, or in the controls. In psoriatic arthritis, however, these levels were significantly higher only in those patients with peripheral joint disease and not in those with axial arthritis alone. There was no significant increase in antibody levels in patients with AS or Reiter's syndrome. There were no differences in the levels of IgG or IgM antibodies to CK-18 or EpK between the patient groups and controls.. Raised levels of IgA antibodies to CK-18 and EpK in psoriatic arthropathy and psoriasis probably reflect exposure of intracellular cytokeratin antigens to the immune system after damage to cytokeratin containing cells, and suggests a common pathogenic mechanism in these conditions which involves production of cytokeratin autoantibodies. In patients with psoriatic arthropathy, such a mechanism appears only to be operating in patients with peripheral joint involvement and not in those with axial arthritis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Arthritis, Psoriatic; Arthritis, Reactive; Autoantibodies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunoglobulin A; Immunoglobulin G; Keratins; Male; Middle Aged; Psoriasis; Skin; Spondylitis, Ankylosing

The morphology of hair follicles was examined in psoriatic scalp biopsies and compared with normal scalp. In scalp psoriasis the lower outer root sheath and hair matrix were not affected by the psoriatic changes, although there was an irregular expansion in the proximal lower outer root sheath. This area has been characterized, by the presence of keratin K19-containing cells, as the putative stem cell region. In addition, marked shrinkage of the sebaceous glands was seen in the psoriatic scalp, as previously reported. A panel of monospecific monoclonal antibodies to individual epithelial keratins was used to analyse scalp specimens immunohistochemically. Keratin expression in scalp was generally unaffected by psoriasis, except for widespread expression of suprabasal keratins K16 and K17 in suprabasal interfollicular psoriatic scalp epidermis. Simple epithelial keratins K8 and K18 were not found in follicular epithelium from either normal or psoriatic scalp, using multiple monospecific antibodies. This study shows that keratin K17 is induced suprabasally during epidermal hyperproliferation, and cannot therefore be considered a hair follicle-specific keratin.

Topics: Adolescent; Adult; Cell Differentiation; Child; Child, Preschool; Epithelium; Female; Humans; Immunohistochemistry; Infant; Keratinocytes; Keratins; Male; Middle Aged; Psoriasis; Scalp; Scalp Dermatoses; Sebaceous Glands

We have investigated markers of epidermal proliferation and differentiation in terms of keratin expression, the morphology of the cutaneous vasculature, and numbers of cutaneous mast cells, in patients with chronic plaque psoriasis. Using the phenomenon of the 'active edge', we have studied these features in the psoriatic plaque itself, and in the clinically normal active and inactive edges of the same plaque. Our results confirm the anticipated changes in keratin profiles, mast cell numbers and psoriatic morphology of the vasculature within the plaque itself. They further indicate that the vascular changes precede the epidermal and mast cell features at the active edge, and that the inactive edge is inactive for all of these variables. Mediators responsible for the vascular proliferation and elongation must be present in increased amounts at the active edge when compared with the inactive, and include locally produced and circulating factors.

Topics: Cell Count; Electrophoresis, Polyacrylamide Gel; Epidermis; Humans; Keratins; Mast Cells; Psoriasis

Nail pathology shares some common features with skin pathology, but it also has its own peculiar aspects. The anatomical and physiological characteristics of the nail unit probably play a major role in determining these pathological differences. Although the presence of keratohyaline granules is a normal feature of the skin, there is no granular layer in the normal nail matrix. As a consequence, nail matrix hypergranulosis should be considered a separate entity from skin hypergranulosis. In our review of 150 longitudinal nail biopsy specimens, keratohyaline granules were seen in the nail matrix of 24 cases of lichen planus, 29 cases of spongiotic trachyonychia, 10 cases of psoriasis, and three cases of Hallopeau acrodermatitis. In all cases, the presence of keratohyaline granules was associated with the absence of the normal keratogenous zone. Similar nail matrix features were detectable in three cases of malignant melanoma, two cases of primary systemic amyloidosis, and one case of histiocytoid hemangioma compressing the nail matrix. Our data suggest that inflammatory and compressive insults to the nail matrix cause both disappearance of the keratogenous zone and matrix keratinization with the formation of keratohyaline granules. Skin hypergranulosis reflects a hyperplasia of a normal skin component. In the nail matrix, however, hypergranulosis represents the appearance of structures not normally present. Nail matrix hypergranulosis should be considered a pattern of nail matrix reaction to different inflammatory insults. It is therefore more analogous to epidermal parakeratosis than to epidermal hypergranulosis.

Topics: Acrodermatitis; Amyloidosis; Biopsy; Epidermis; Epithelium; Hemangioma; Humans; Hyalin; Hyperplasia; Keratins; Keratosis; Lichen Planus; Melanoma; Nail Diseases; Nails; Onychomycosis; Psoriasis

The morphological and biochemical characteristics of psoriasis are well documented, but the pathogenesis of this disease is not clearly understood. A variety of in vitro models of psoriasis have been developed in attempts to identify the trigger factors, but no model so far reproduces the stable psoriatic phenotype accurately. In the present work, we initially checked the immunohistochemical distribution of proliferation/differentiation markers in psoriatic skin in vivo, and our results largely confirm previously reported data. However the study was performed using a new series of monoclonal antibodies to keratin. Subsequently we took normal or psoriatic skin biopsies, reconstructed skin equivalents using a recently described model and analysed the proliferation/differentiation status of the resulting epidermis. Dramatic morphological and antigenic differences were found between normal and psoriatic skin in vivo, but whatever the source of the initial biopsy, a unique in vitro phenotype was obtained in the reconstructed epidermis. This phenotype was marked by mild hyperproliferation and an altered distribution of differentiation-associated antigens suggesting a need for extracutaneous stimuli to maintain the psoriatic phenotype in vitro.

Topics: Antibodies, Monoclonal; Antibody Specificity; Biomarkers; Cell Differentiation; Cell Division; Cells, Cultured; Epidermal Cells; Epidermis; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Keratins; Psoriasis; Sensitivity and Specificity; Skin

Topics: Bandages, Hydrocolloid; Biomarkers; Colloids; Epidermis; Filaggrin Proteins; Humans; Intermediate Filament Proteins; Keratins; Occlusive Dressings; Protein Precursors; Psoriasis; Skin

A murine monoclonal antibody, FB1, reacted with the basal keratinocytes of human stratified epithelia. One-dimensional and two-dimensional immunoblotting assays, performed on keratins extracted from HaCat cells and normal human keratinocytes, showed that FB1 recognizes K14. When LL002, another K14 monoclonal antibody is added, the FB1 stained area in the 2D-immunoblot seems to cover a fraction of the LL002 spot. Immunohistochemical data obtained from studies on normal human tissues supported the K14 specificity of FB1, but when compared with two other monoclonal antibodies, LL002 and RCK107 reacting with K14, some differences appeared. These differences were mainly seen in sweat glands, hair follicles, psoriatic epidermis and salivary glands. In psoriatic epidermis, FB1 showed a heterogeneous pattern of staining of the basal cell compartment. Intense reactivity was only observed at the bottom of the rete ridges. Staining diminished and finally disappeared in the basal cells above the dermal papillae. This observation supports the view that an increased germinative cell population in psoriasis involves a partially differentiated amplifying compartment in which the number of cell divisions is increased.

Topics: Animals; Antibodies, Monoclonal; Epitopes; Humans; Immunohistochemistry; Keratin-14; Keratinocytes; Keratins; Microscopy, Immunoelectron; Psoriasis; Reference Values; Staining and Labeling

The topical application of a hydrocolloid occlusive dressing (HCD) has been shown in various studies to have an antipsoriatic effect as monotherapy but especially in combination with a topical corticosteroid. The aim of the present study was to assess the effect of 3 weeks of HCD monotherapy at the immunohistochemical level. Ten patients were treated. Before and after treatment, a biopsy was taken, and immunohistochemical stainings were carried out with markers for epidermal growth, keratinization, inflammation and endothelium. Suprabasal expression of keratin 16, the number of cycling epidermal cells and the number of polymorphonuclear leucocytes and T lymphocytes tended to decrease during treatment. The endothelial markers did not change during HCD treatment. This study confirms the antipsoriatic effect of HCD and demonstrates that its effect upon some markers of inflammation, epidermal proliferation and keratinization is modest.

Topics: Adult; Aged; Bandages, Hydrocolloid; Cell Cycle; Colloids; Endothelium, Vascular; Epidermis; Erythema; Female; Humans; Immunohistochemistry; Keratins; Male; Middle Aged; Neutrophils; Occlusive Dressings; Psoriasis; Staining and Labeling; T-Lymphocytes

In the human hair follicle, outer root sheath (ORS) cells constitutively express the hyperproliferation-associated keratins 6, 16 and 17 instead of keratins 1 and 10 found in interfollicular epidermis. In organotypic cultures. ORS cells form a stratified epithelium which in many respects resembles psoriatic skin: it has a hyperplastic tissue architecture and a poorly developed granular layer, and expresses hyperproliferation-associated keratins. Therefore, we studied the effects of the antipsoriatic compounds 1 alpha,25-dihydroxy-vitamin D3 (1 alpha,25-(OH)2-D3) and its synthetic derivative calcipotriol on cultured ORS cells. In monolayer cultures, 10(-6) M 1 alpha,25-(OH)2-D3 or calcipotriol completely blocked ORS cell proliferation. This inhibitory effect was substantially reduced at 10(-8) M. Incubation of organotypic ORS cultures with both vitamin D analogues resulted in a marked thinning of the living cell compartment concomitant with a thickening of the horny layer. A reduced expression of differentiation markers such as keratins 10, 16 and 17, involucrin and filaggrin paralleled the thinning of the stratum Malpighi. As determined by quantification of BrdU-positive cells, ORS cell proliferation was apparently not affected by the vitamin D analogues, indicating that these compounds mainly operate by accelerating the differentiation pathway within the suprabasal living cell compartment. No alteration in the expression of the alpha 6- and beta 1-integrin chains was found.

Topics: Adult; Calcitriol; Cell Differentiation; Cell Division; Dermatologic Agents; Filaggrin Proteins; Hair; Humans; Keratins; Models, Biological; Organ Culture Techniques; Psoriasis

Monospecific antibodies to individual keratin polypeptides can be used to examine the tissue and cellular coexpression of members of keratin pairs. Monospecific monoclonal and polyclonal antibodies have been raised to keratins 1 and 10 using both crude cytoskeletal extracts and synthetic peptides. The tissue distribution of these keratins has been determined against a panel of freshly frozen normal tissues from humans, rodents and pigs. Epidermal expression has been examined in psoriatic plaques, and healing wounds, as examples of epidermal hyperproliferation. Cultured keratinocytes in monolayer (low calcium), stratified (high calcium), and complex cultures, transformed keratinocytes, and tumour cell lines, have been examined for the in vitro expression of these keratins. The sensitivity and precise localization of reactivity with these monospecific antibodies gives a highly accurate picture of individual cell expression. There is confirmation of coexpression of keratins 1 and 10 in epidermal and mucosal sites, and with keratin 16 in hyperproliferative states. These monospecific antibodies provide an important means of examining keratin expression in epidermal tumours and keratinizing disorders, and of seeking keratin mutations in cell lines and in skin diseases.

Topics: Adult; Animals; Antibodies, Monoclonal; Biomarkers; Cell Differentiation; Cell Division; Cell Line; Cells, Cultured; Epidermal Cells; Epidermis; Gene Expression Regulation; Humans; Infant, Newborn; Keratinocytes; Keratins; Keratosis; Mice; Mice, Inbred BALB C; Mucous Membrane; Psoriasis; Skin; Skin Neoplasms; Swine; Wound Healing

We examined the expression of the transglutaminase 1 (TGase 1) gene in frozen sections of normal and psoriatic epidermis by means of non-radioactive in situ hybridization with digoxigenin-labelled cRNA probes. TGase 1 mRNA was expressed in the granular layer of normal epidermis, regardless of ortho- or hyperkeratosis. However, in psoriatic epidermis, TGase 1 mRNA was detected in the suprabasal spinous layer, but not in the subcorneal layer. These results indicate that TGase 1 gene expression is limited to the last stage of keratinization in normal epidermis and this regulation is disturbed in psoriasis.

Topics: Adult; Cell Differentiation; Gene Expression Regulation, Enzymologic; Humans; In Situ Hybridization; Keratins; Male; Psoriasis; RNA, Messenger; Skin; Transglutaminases

In this study we define the proliferative compartments of in vivo human epidermis, using specific antibodies related to cell differentiation (beta 1 and beta 4 integrins and K1/K10 differentiation keratins) and cell cycle (proliferating cell nuclear antigen [PCNA]) in combination with flow cytometric quantitation of the DNA content and optical characteristics of the cells. The beta 1 integrin (CD29) marked both of the potentially proliferative subsets in normal epidermis. One subset of normal epidermis is CD29+ K1/K10-, which was predominantly basal, and found to be comprised of slow cycling, small cells with primitive cytoplasmic organization. The vast majority (95.5%) of these cells were in a quiescent state (G0/early G1) as indicated by their lack of the cyclin, PCNA. The other proliferative subset of normal epidermis was CD29+ K1/K10+, which was suprabasal and occasional basal, highly proliferative, larger in size, and which exhibited a more complex cytoplasmic structure. Because early differentiation (K1/K10 expression) has begun in the CD29+ K1/K10+ subset, it is highly likely that they represent the proliferative population which is capable of transiently amplifying itself before terminal differentiation. Within lesional psoriatic epidermis, similar proliferative cell populations were present as in normal epidermis, and the hyperproliferative defect was localized to the beta 1 and beta 4 integrin+, K1/K10- populations, which in normal epidermis is basally located and quiescent with regard to cell cycle. In psoriatic epidermis, a six- to sevenfold increase in the number of cells in the S/G2+M phase of cell cycle was found among CD29+ K1/K10- cells (p < 0.05). Furthermore, all lesional K1/K10- cells showed high PCNA positivity, indicating that all these cells had been recently induced into cell cycle. By contrast, the proportion of cycling cells among lesional psoriatic CD29+ K1/K10+ keratinocytes was similar to normals. Anti-HLA-DR, CD45, and vimentin antibodies were used to concomitantly track the proliferative states of Langerhans cell, melanocyte, and infiltrating leukocyte populations. In normal epidermis, the cycling fractions (cells in S/G2/M phase) of these cells were similar to the CD29+K1/K10- keratinocytes, whereas in lesional epidermis their cycling pools were increased relative to normal, but not so much as the proliferative fractions of psoriatic CD29+ K1/K10- keratinocytes. These data demonstrate the use of simultaneous analysis of int

Topics: Antigens, CD; Autoantigens; Cell Division; Epidermis; Flow Cytometry; HLA-DR Antigens; Humans; Integrin beta1; Integrins; Keratinocytes; Keratins; Leukocyte Common Antigens; Light; Nuclear Proteins; Proliferating Cell Nuclear Antigen; Psoriasis; Scattering, Radiation; Vimentin

Topics: Animals; Autoantibodies; Dermatitis, Contact; Dogs; Female; Guinea Pigs; Humans; Immunohistochemistry; Keratins; Male; Psoriasis; Rabbits; Skin

Etretinate, a second generation retinoic acid, has been reported to be useful in the treatment of psoriasis and other keratinizing disorders. The effectiveness of etretinate for these disorders are studied in a 10-year retrospective study of all patients treated with etretinate in a skin clinic in Singapore.. The case records of 190 cases of psoriasis and other keratinizing disorders treated with etretinate were analyzed. Information collected included demographic data, dosage of etretinate taken, response and side effects, clinical follow-up, and relapse.. Most of the cases (72.6%) had psoriasis (138/190). The others had different keratinizing disorders. The dose of etretinate used was 0.15 to 1 mg/kg/day (median 0.36 mg/kg/day), and the duration of the treatment varied from 1 to 120 months (median 6 months). Etretinate was coadministered with UVB (ReUVB) or PUVA (RePUVA) in 89 (46.8%) patients. In psoriasis, the response to treatment was excellent in 41.3% (57/138), good in 36.2% (50/138), fair in 15.9% (22/138), and poor in 6.5% (9/138) of the cases. Patients with plaque-type psoriasis did better with combination therapy than with monotherapy. Those with keratinizing disorders showed excellent, good, fair, and poor responses in 32.7% (17/52), 32.7% (17/52), 25.0% (13/52), and 9.6% (5/52) respectively. Adverse effects were noted in 102 (53.7%) cases and were generally mild and tolerable. Etretinate was discontinued in 24 (12.6%) patients due to significant toxicity.. Etretinate is effective for treating psoriasis and other keratinizing disorders. Combination therapy is preferred in chronic plaque psoriasis. Adverse effects are common, but mild and tolerable.

Topics: Adolescent; Adult; Child; Etretinate; Female; Humans; Keratins; Male; Middle Aged; Psoriasis; Retrospective Studies; Skin Diseases

Topics: Adolescent; Adult; Aged; Antibodies; Enzyme-Linked Immunosorbent Assay; Female; Focal Infection; Humans; Immunoglobulin G; Immunoglobulin M; Keratins; Male; Middle Aged; Psoriasis; Staphylococcal Infections; Streptococcal Infections; Tonsillectomy; Tonsillitis

The skin equivalent (SE) has been validated as a model for studies on proliferation of keratinocytes. SEs were prepared from normal skin by implanting punch biopsies on dermal equivalents consisting of fibroblasts in a collagen matrix. The outgrowths were measured by planimetry. An immunohistochemical investigation with antibodies against markers associated with proliferation was performed on frozen sections from SEs during outgrowth at 3-6 days (SE6) as well as after completion of outgrowth at 21 days (SE21). Biopsies from normal controls and from uninvolved and involved skin in psoriatic patients were also studied. The antibodies used were Ki-67, cytokeratin 8.12, and antibodies against the receptors for epidermal growth factor (EGFr) and transferrin (TFr). The increase in area was linear during the first 7 days of culture and usually reached the edges of the dermal equivalent at this time. In SE6 TFr was expressed in the basal part of the outgrowth while the other markers were not observed. In SE21 and in psoriasis there was abundant epidermal staining of Ki-67-positive nuclei and cytokeratin 8.12 was detected in the suprabasal part of the epidermis. EGFr and TFr were seen in the basal layer in SE21. In the psoriatic lesions these receptors were found both in the basal and suprabasal layers. The lack of proliferation markers in SE6 indicates that the initial increase in the area of keratinocytes is due to migration from the punch biopsies. Increased cell proliferation is present in SE21, a finding in common with psoriasis and wound healing. The skin equivalents should therefore be an appropriate model for studies on these phenomena.

Topics: Adult; Aged; Cell Division; Cells, Cultured; ErbB Receptors; Female; Humans; Immunohistochemistry; Keratinocytes; Keratins; Ki-67 Antigen; Male; Middle Aged; Neoplasm Proteins; Nuclear Proteins; Psoriasis; Receptors, Transferrin; Skin

Three biopsies of normal skin and 15 biopsies collected from patients with psoriasis vulgaris were analyzed for the expression of the 50 kd cytokeratin using direct immunofluorescence and ABC technique before and after local treatment with anthralin 0.1% in a petrolatum base, with 0.05% betamethasone dipropionate cream, and finally, after PUVA treatment. Antiserum against the 50 kd anti-cytokeratin reacted with tissue sections of normal skin, staining cells in the basal layer, while the psoriatic skin sections before the various treatments showed a staining concerning the whole thickness of the epidermis. After the various therapies, the 50 kd cytokeratin immunoreactivity was observed only in the basal layer of those psoriatic skin sections that showed complete clinical clearing, while it was observed in the whole thickness of psoriatic patches that did not clear. These data suggest that the normalization of the 50 kd cytokeratin expression pattern can be considered as a marker of clinical remission of psoriasis.

Topics: Administration, Topical; Adolescent; Adult; Anthralin; Anti-Inflammatory Agents; Betamethasone; Epidermis; Female; Fluorescent Antibody Technique; Glucocorticoids; Humans; Immunoenzyme Techniques; Keratins; Male; Methoxsalen; Middle Aged; Molecular Weight; Psoriasis; PUVA Therapy; Skin

Cyclosporine (CSA) decreases lymphokine synthesis and keratinocyte proliferation in vitro, but its in vivo mechanism of action in treating recalcitrant psoriasis is incompletely understood. Ten psoriasis patients were treated with CSA (2-7.5 mg/kg/d) with clinical improvement in nine of 10 patients. Skin biopsies before and after 1-3 months of CSA treatment were studied for evidence of immune and keratinocyte activation using immunoperoxidase and Northern blotting analysis. The number of activated, IL-2 receptor+ T cells in plaques after CSA treatment was reduced in all patients by a mean of 60%. Seven of 10 patients showed a decrease in keratinocyte HLA-DR expression; five of seven showed a decrease in gamma-IP-10 immunoreactivity, suggesting a decline in gamma interferon levels in plaques after CSA therapy. We studied the effect of CSA treatment in vivo on TGF-alpha, IL-6, and keratin K16 expression, three markers of keratinocyte growth activation. Expression of keratinocyte TGF-alpha and IL-6, which are elevated in active psoriatic epidermis, did not change in these patients after CSA treatment. The majority of patients (five of eight) continued to express the hyperproliferative keratin K16 after CSA treatment. Our results suggest that the predominant direct mechanism of action of Cyclosporine in vivo is a diminution of T-cell activation in plaques, with attendant decreased lymphokine production.

Topics: Cyclosporine; HLA-DR Antigens; Humans; Interleukin-6; Keratins; Lymphocyte Activation; Psoriasis; Receptors, Interleukin-2; Regeneration; Skin; T-Lymphocytes; Transforming Growth Factor alpha

In normal epidermis, as previously reported, the first signs of differentiation occur within the basal layer in a subpopulation of keratinocytes that start to express K1 and K10 "supra-basal" keratin transcripts (20-30% of the basal cells) and proteins (5-10% of the basal cells). We found that in psoriatic lesions, the basal layer was devoid of cells expressing these early differentiation markers. This was already the case at the periphery of the lesions, where epidermis, although slightly acanthotic, still completes the keratinization process. In the center of the lesions, not only the basal layer, but also several rows of suprabasal cells, were negative for keratin K10 transcripts or protein. Moreover, the upper nucleated layers of involved epidermis were also devoid of K10 keratin transcripts or proteins. In normal epidermis, as previously reported, transcripts for the "basal" K5 keratin were mainly restricted to the basal layer, whereas the protein persisted in a few suprabasal layers. We found that in psoriatic epidermis, K5 keratin transcripts persisted in several suprabasal layers up to the level where K10 keratin transcripts appeared. These data, although not contradictory with previous reports showing a reduction of K1-K10 keratins and other differentiation markers in psoriasis, demonstrate that these quantitative changes are in fact the result of major qualitative differences in the distribution of these markers in psoriatic versus normal skin. Our results indicating that the onset of differentiation is delayed in psoriasis show that, contrary to conclusions accepted so far, not only the suprabasal compartment, but also the basal one, is abnormal in psoriatic epidermis.

Topics: Cell Differentiation; Epidermis; Humans; Keratins; Protein Precursors; Psoriasis; RNA Probes; RNA, Messenger

Human epidermal cell cultures were used to study the effects of retinoids on keratinocyte differentiation. Keratin profiles were studied by quantitative gel electrophoresis of culture extracts, whereas the extent of envelope formation was assessed in an enzyme-linked immunosorbent assay (ELISA) using an antibody that specifically recognizes keratinocyte envelopes. Exposure of cultures to a variety of different retinoids produced both dose-dependent decreases in keratin 16 with consequent increases in the keratin 14: keratin 16 ratio, and a decrease in envelope formation. The order of activity in both assays was similar: arotinoid ethyl ester (Ro 13-6298) greater than or equal to arotinoid acid (Ro 13-7410) much greater than all trans retinoic acid (Ro 1-5488) greater than acitretin (Ro 10-1670) greater than or equal to etretinate (Ro 10-9359), the only difference being that acitretin was slightly more active than etretinate in the keratin assay whereas these retinoids were equi-active in the envelope assay. Analysis of the lesional keratins of psoriasis patients showed that etretinate caused a reduction in keratin 16 and an increase in the keratin 14:keratin 16 ratio, although the magnitude of these changes and their correlation with clinical improvement was variable. As the in vitro assays reported here are simple and quick, they allow rapid screening of compounds for retinoid-like activity.

Topics: 3T3 Cells; Animals; Biopsy; Cell Differentiation; Cell Membrane; Etretinate; Growth; Humans; Keratinocytes; Keratins; Mice; Psoriasis; Retinoids; Skin

Molecular mimicry between mycobacterial heat-shock protein (hsp) 65 and host tissue antigens have been implicated in the autoimmune pathogenesis of certain idiopathic diseases. Here, we demonstrated that two of our previously characterized monoclonal antibodies (mAb), Ne5 and Nd4 that were directed to a carboxy-terminal epitope on the mycobacterial hsp 65, specifically cross-reacted with suprabasal cytokeratin of the normal human skin. These mAb also showed similar keratin staining of hair follicle epithelia and produced no reaction with other dermal components. Both mAb strongly stained the cytoplasm of the majority of freshly isolated epidermal keratinocytes from the normal human skin. None of these mAb showed staining with human HeLa cells and with human skin fibroblasts. Immunoblotting using total keratin extract prepared from isolated epidermal keratinocytes revealed that mAb Ne5 and Nd4 specifically reacted with a molecular size of 65,000-67,000 MW keratin protein(s) and such reactivity was not observed from cytoskeletal proteins extracted from HeLa cells and skin fibroblasts. Comparison of immunoblotting reactivity with conventional anti-cytokeratin mAb further revealed that mAb Ne5/Nd4 recognized a 65,000-67,000 MW molecular-sized protein corresponding to cytokeratin 1/2 from the same keratinocyte extract as anti-cytokeratin mAb. Preincubation of mAb Ne5/Nd4 with the purified mycobacterial hsp 65 abolished this keratin cross-reactivity in both immunohistochemistry and immunoblotting. Moreover, these mAb showed no keratin staining in lesional psoriatic skin and also reacted weakly with cultured epidermal keratinocytes. Since mAb Ne5/Nd4 specifically recognized a 67,000-65,000 MW molecular-sized protein(s) derived from epidermal keratinocytes and the known characteristics of epidermal cytokeratin 1/2 appeared to be consistent with present results, we concluded that Ne5/Nd4 cross-reactive protein(s) in the human epidermis is suprabasal cytokeratin 1/2. Comparison of the previously mapped Ne5/Nd4 epitope region of amino acid residues 525-540 of the mycobacterial hsp 65 with the entire sequence of human 65,000 MW keratin revealed that a stretch of nine amino acids of the Ne5/Nd4 epitope sequence resembled certain regions of the carboxy-terminus of the human 65,000 MW keratin. This similarity of the mycobacterial hsp 65 probably contributes to the cytokeratin cross-reactive epitope. Our results presented here demonstrate direct evidence of immunological

Topics: Amino Acid Sequence; Antibodies, Monoclonal; Antigens, Bacterial; Cells, Cultured; Cross Reactions; Epidermis; Epitopes; Heat-Shock Proteins; Humans; Immunoenzyme Techniques; Keratins; Molecular Sequence Data; Mycobacterium tuberculosis; Psoriasis

Transforming growth factor-alpha (TGF-alpha) is thought to be the major autocrine factor controlling growth in epidermal cells. To explore further the role of TGF-alpha in epidermal growth and differentiation, we used a human keratin K14 promoter to target expression of rat TGF-alpha cDNA to the stratified squamous epithelia of transgenic mice. Unexpectedly, the only regions of epidermis especially responsive to TGF-alpha overexpression were those that were normally thick and where hair follicle density was typically low. This included most, if not all, body skin from 2-day- to 2-week-old mice, and ear, footpad, tail, and scrotum skin in adult mice. In these regions, excess TGF-alpha resulted in thicker epidermis and more stunted hair growth. Epidermal thickening was attributed both to cell hypertrophy and to a proportional increase in the number of basal, spinous, granular, and stratum corneum cells. During both postnatal development and epidermal differentiation, responsiveness to elevated TGF-alpha seemed to correlate with existing epidermal growth factor (EGF) receptor levels, and we saw no evidence for TGF-alpha-mediated control of EGF receptor (EGFR) expression. In adults, no squamous cell carcinomas were detected, but benign papillomas were common, developing primarily in regions of mechanical irritation or wounding. In addition, adult transgenic skin that was still both sensitive to TGF-alpha and subject to mild irritation displayed localized regions of leukocytic infiltration and granular layer loss, characteristics frequently seen in psoriasis in humans. These unusual regional and developmental effects of TGF-alpha suggest a natural role for the growth factor in (1) controlling epidermal thickness during development and differentiation, (2) involvement in papilloma formation, presumably in conjunction with TGF-beta, and (3) involvement in psoriasis, in conjunction with some as yet unidentified secondary stimulus stemming from mild mechanical irritation/bacterial infection.

Topics: Animals; Animals, Newborn; Blotting, Northern; Cell Differentiation; Cell Division; Cells, Cultured; Epidermis; ErbB Receptors; Humans; Keratins; Mice; Mice, Transgenic; Papilloma; Phenotype; Psoriasis; Transforming Growth Factor alpha; Wounds and Injuries

The immunohistochemical distribution of the epidermal proteins filaggrin, involucrin, and cytokeratins is characteristic in normal epidermis. This distribution may change as a result of malignant transformation or abnormal differentiation. The present study was conducted to determine the patterns of reactivity of psoriatic epidermis to antibodies against various epidermal proteins and to clarify abnormal differentiation or maturation of the keratinocytes in psoriatic epidermis. Anti-human filaggrin, anti-human involucrin, and twelve kinds of anti-cytokeratin antibodies were used in this study. Cryostat or paraffin-embedded sections were stained with these antibodies by the avidin-biotin peroxidase technique. The epidermis of the noninvolved skin of patients with psoriasis vulgaris showed the distribution seen in normal skin. However, involved psoriatic skin revealed little or no reaction in the stratum corneum or in the granular layer with the anti-filaggrin antibody. Cells positively staining with anti-involucrin antibody paradoxically appeared in the lower cell layers of involved psoriatic epidermis. An anti-keratin antibody, AE1, stained suprabasal cells in involved psoriatic epidermis, although this antibody selectively stained epidermal basal cells in normal skin. The other anti-keratin antibodies, especially KL1, PKK1, and a polyclonal anti-keratin antibody, were less reactive with involved psoriatic skin than with normal skin. These observations suggest that the maturation pathway of keratinocytes in active psoriatic lesions differs qualitatively from that in normal epidermis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Autoantibodies; Cell Differentiation; Epidermis; Female; Filaggrin Proteins; Humans; Immunohistochemistry; Intermediate Filament Proteins; Keratins; Male; Middle Aged; Protein Precursors; Psoriasis; Skin

Inflammatory Linear Verrucous Epidermal Nevus (ILVEN) has been suggested to be a separate disease entity. However, the distinction from linear psoriasis has been discussed in the literature over recent decades. The aim of the present study was to investigate, in addition to the clinical and histological criteria, the immunohistochemical aspects of inflammation, epidermal proliferation and keratinization. From a clinical and histological point of view, ILVEN and psoriasis, according to the established criteria, have been proved to overlap. The immunohistochemical study suggests that the following procedures have an additional diagnostic impact: assessment of elastase-positive cells, assessment of keratin 16 and of keratin 10.

Topics: Adult; Aged; Antibodies, Monoclonal; Chromoblastomycosis; Diagnosis, Differential; Humans; Keratins; Keratosis; Male; Middle Aged; Nevus; Pancreatic Elastase; Psoriasis; Skin Neoplasms

Psoriasis can be triggered by haemolytic streptococcal infections. As M protein is a major pathogenic surface antigen in these streptococci, the cross-reactivity between streptococcal M protein surface antigens and human epidermis was investigated. The conserved component common to the few M proteins investigated consists of an alpha-helical 'coiled-coil' configuration, similar to sub-units of human keratin. The amino acid sequence of protein M6, one of the M proteins that has been fully sequenced, was compared with that of 4721 ubiquitous peptides, by computer-assisted analysis using a protein-sequence data bank. Of all human proteins in the data bank 50-kDa keratin type 1 showed the closest homology with protein M6. Further evaluation revealed that this homology mainly involved the heptapeptide repeat patterns, which form the alpha-helical 'coiled-coil' structure, in both M6 and 50-kDa keratin. Cryostat sections of normal, involved and uninvolved psoriatic skin were studied for cross-reactivity with rabbit antisera raised against 10 different M proteins. All these antisera reacted with the stratum corneum of normal and psoriatic epidermis to a variable extent. Staining of keratinocyte cytoplasm was also observed, but this tended to be more prominent in lesional than in uninvolved and normal skin. Some of the M antisera also stained dendritic cells in the upper dermis as well as endothelium and smooth muscle. These cross-reactivities might be relevant to the pathogenesis of post-streptococcal psoriasis.

Topics: Amino Acid Sequence; Antigens, Bacterial; Antigens, Surface; Bacterial Outer Membrane Proteins; Bacterial Proteins; Carrier Proteins; Cross Reactions; Electronic Data Processing; Epidermis; Fluorescent Antibody Technique; Humans; Keratins; Molecular Sequence Data; Psoriasis; Sequence Alignment

A monoclonal antibody (ES3A) was raised against a mouse graft-versus-host reaction (GVHR) model. This antibody was against basal cell cytoplasm and reacted with an acidic (pI 6.2) 50 kDa keratin of human epidermis. However, ES3A reacted with several lower layers of epidermal cells in psoriasis and seborrhoeic keratosis. Acanthotic seborrhoeic keratosis showed varying patterns even in a single lesion. If combined with FACS analysis, ES3A-positive cells could be quantified. Normal skin showed 28%, while psoriasis and seborrhoeic keratosis showed 44% and 51%, respectively. ES3A-positive compartments of the acanthotic type of seborrhoeic keratosis were larger than those of the hyperkeratotic type. ES3A may be suitable for quantification of germinative or proliferative cells.

Topics: Animals; Antibodies, Monoclonal; Cell Fusion; Cell Separation; Dermatitis, Seborrheic; Disease Models, Animal; Epidermal Cells; Flow Cytometry; Fluorescent Antibody Technique; Graft vs Host Reaction; Humans; Immunoblotting; In Vitro Techniques; Keratins; Keratosis; Mice; Propidium; Psoriasis

Topics: Anthralin; Antibodies, Monoclonal; Biomarkers; Calcitriol; Frozen Sections; Gene Expression; Humans; Immunohistochemistry; Keratins; Psoriasis; Skin

Keratin expression in lesional, marginal and uninvolved psoriatic epidermis was analysed by one- and two-dimensional gel electrophoresis and immunoblotting. Keratins K1, K5, K6, K10, K14, and K16 were identified in lesional epidermis. Keratins K6 and K16 were found in all epidermis probes of uninvolved skin, but never occurred in normal epidermis of control skin samples. By means of laser-densitometric evaluation of one-dimensional gels a downregulation of K1 and K10 and an upregulation of K6 and K16 was found in psoriatic epidermis. Unexpectedly, the level of K5 was considerably lower and the level of K14 considerably higher in lesional skin than in normal epidermis. These results demonstrate that not only basal keratinocytes in lesional epidermis but also suprabasal keratinocytes in uninvolved psoriatic epidermis express an altered differentiation pattern. The latter phenomenon could be very important in understanding the development of the so-called "Köbner effect" in psoriatic epidermis.

Topics: Adult; Aged; Aged, 80 and over; Biopsy; Cell Division; Cells, Cultured; Densitometry; Electrophoresis, Gel, Two-Dimensional; Epidermis; Female; Gene Expression; Humans; Immunoblotting; Keratins; Lasers; Male; Middle Aged; Psoriasis

To study the development of the psoriatic lesion, biopsies were taken from the margin of spreading plaques and acute pinpoint papules. Consecutive sections across the margin were stained using different monoclonal antibodies to characterize epidermal growth (Ki-67) and abnormal keratinization (Ks8.12, RKSE60). All three immunohistochemical markers showed pronounced changes in the lesional skin with a clear transition to the uninvolved skin. The suprabasal Ks8.12 binding was the earliest change found in the epidermis, and its localization high in the suprabasal compartment indicates that metabolic dysregulation in this cell population was not a consequence of the recruitment process in the basal layer.

Topics: Adult; Biomarkers; Cell Division; Female; Humans; Keratins; Male; Middle Aged; Psoriasis; Skin; Staining and Labeling

The in vivo expression of the proliferation keratins K 6 and K 16 is characteristic for hyperproliferative diseases and positive in psoriatic lesions. Our study was performed in order to investigate the in vitro expression of proliferation keratins and to settle the question whether they are also inducible in organ cultures of uninvolved psoriatic skin, possibly differing from normal skin.

Topics: Adult; Aged; Aged, 80 and over; Biopsy; Cell Division; Female; Humans; Keratins; Male; Middle Aged; Peptides; Psoriasis; Skin

Using the monoclonal antibody Ks8.12 directed against keratins 13 and 16, we have studied psoriatic and trauma-induced hyperproliferation. In normal skin 40 and 64 h after sellotape stripping and in psoriatic lesions, a pronounced suprabasal staining by Ks8.12 was observed. In unstripped normal skin and in the clinically uninvolved skin of psoriatic patients some patchy staining limited to the basal layer was seen. The clear distinction between normal and hyperproliferative skin in keratin expression as detected by Ks8.12 indicates that this antibody may serve as a marker for epidermal proliferation in psoriasis.

Topics: Antibodies, Monoclonal; Epidermis; Humans; Immunohistochemistry; Keratins; Psoriasis; Time Factors

Psoriasis occurring in a patient with lamellar ichthyosis is reported. Similar self-limited episodes had occurred earlier. On histopathologic examination of a biopsy specimen, an eruptive plaque showed parakeratosis and a reduction in the granular layer. Electrophoretic analysis of the keratins isolated from the epidermis of a plaque showed a reduction in the amount of the 67 kd polypeptide compared to the keratins of ichthyosis epidermis. Both of these findings support the diagnosis of psoriasis. Epilyt, applied daily, was effective in removing scales.

Topics: Adult; Biopsy; Dermatologic Agents; Female; Humans; Hyperplasia; Ichthyosis; Keratins; Psoriasis; Skin

Studies on the anti-keratin intermediate filament autoantibodies (anti-KIF-Abs) in sera from psoriasis (Pso) patients were performed by immunoblotting and enzyme-linked immunosorbent assay (ELISA). According to their reactivities against keratin subunits (50, 56.5, 58, and 63-68 kd), sera were divided into four groups. However, no significant differences in these reactive patterns were found between healthy volunteers and Pso patients. In the second experiment, anti-KIF-Abs in sera from Pso patients, pustulosis palmaris et plantaris (PPP) patients, atopic dermatitis (AD) patients, systemic lupus erythematosus (SLE) patients, and healthy volunteers were determined by ELISA, using as substrate keratins purified from normal human stratum corneum and 48- and 50-kd keratins purified from psoriatic stratum corneum. The serum titers of anti-KIF-Abs against 48- and 50-kd keratins in Pso patients were significantly higher than those in PPP patients, AD patients, SLE patients, or healthy volunteers. The elevated titers of anti-KIF-Abs against the 48- and 50-kd keratins in sera of Pso patients showed a significant decrease with improvement of psoriatic lesions. The above results suggest that anti-KIF-Abs against 48- and 50-kd keratins in sera of Pso patients have some relevance to the severity of the disease and can be used as a marker for the evaluation of the disease activity of psoriasis.

Topics: Autoantibodies; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Immunoblotting; Intermediate Filaments; Keratins; Molecular Weight; Psoriasis; Suppuration

We studied keratin expression in benign epidermal skin diseases, and in Bowen's disease by using three monoclonal cytokeratin antibodies. In adult normal skin, these antibodies bind only to the follicular epithelium (PKK1), the basal keratinocytes (PKK2), or the suprabasal cells in interfollicular epidermis (KA5). Additionally, in fetal epidermis, the PKK1 antibody reacts with basal keratinocytes. In psoriasis and lichen planus, the PKK2 antibody distinctly revealed all epidermal cell layers by immunostaining. However, a negative basal cell-like layer was revealed in both lesions with the KA5 antibody. In pityriasis rubra pilaris, the basal cell layer was uniformly stained with the PKK2 antibody, but only some keratinocytes in upper cell layers showed fluorescence and, in chronic eczema, the 3-4 lowest epidermal cell layers were reactive. The PKK1 antibody did not stain interfollicular keratinocytes in any of the benign proliferative skin diseases studied. In Bowen's disease, a heterogeneous staining pattern with varying intensity among individual cells was seen with all of the antibodies used. Our results suggest different changes in keratin expression in chronic benign and malignant epidermal diseases that may reflect the mechanisms behind these changes.

Topics: Antibodies, Monoclonal; Bowen's Disease; Carcinoma, Squamous Cell; Epidermal Cells; Fluorescent Antibody Technique; Humans; Keratins; Psoriasis; Skin Diseases; Skin Neoplasms

The presence of intercellular adhesion molecule-I (ICAM-I) on keratinocytes of psoriatic skin lesions before and during 8-methoxapsoralen and UVA light (PUVA) treatment was studied. ICAM-I was expressed on the keratinocytes in biopsies of the skin lesions of five patients with psoriasis. The patients who responded to PUVA treatment had a concurrent reduction of ICAM-I expression on the keratinocytes with a reduction of the number of cells in the mononuclear cellular infiltrate and a lessening of the severity of the disease. Patients who went into remission during therapy and then relapsed showed an increase in ICAM-I expression on keratinocytes with an increase in the number of cells in the mononuclear cell infiltrate and an increase in the severity of the disease. HLA-DR expression on keratinocytes was variable during treatment and showed no strong correlation with disease severity.

Topics: Antigens, Surface; Cell Adhesion Molecules; Epidermis; HLA-DR Antigens; Humans; Keratins; Psoriasis; PUVA Therapy; Time Factors

A microprecipitation method was used to test sera of psoriasis patients and control persons for precipitating keratin interfilament antibody (KIF-Ab). Precipitating KIF-Ab were detected in 83% of the psoriasis patients. The sera of only 20.5% of controls without dermatological diseases and 40% of nonpsoriatic patients contained KIF-Ab. The mean KIF-Ab titer of the control and psoriasis group did not differ significantly. The different therapy had different effects on the detectability of precipitating KIF-Ab. Upon completion of dithranol treatment and clinical healing, all sera reacted with KIF from psoriasis scales (pso-sc). PUVA treatment lowered the Ab-titer as well as the number of seropositive sera. These results were confirmed by means of immunoblot and immunodot techniques. Sera from psoriasis patients contained Ab of the IgG and IgM-types against 65, 55 and 45 kD proteins. KIF-IgA-Ab were found frequently in the cases of severe forms of psoriasis.

Topics: Adolescent; Adult; Aged; Autoantibodies; Chemical Precipitation; Female; Humans; Keratins; Male; Middle Aged; Psoriasis

Keratinocyte expression of the monocyte/macrophage surface antigens defined by OKM1 and OKM5 antibodies (Ortho Diagnostics) was examined using the peroxidase anti-peroxidase immunohistochemical technique. A range of inflammatory cutaneous disorders were investigated, including lichen planus, psoriasis and atopic dermatitis. Positive suprabasal keratinocyte expression of OKM5 antigen was observed in all disorders, while keratinocyte staining with OKMI antibody was consistently negative. These results provide further evidence that keratinocytes may play an important role in cutaneous immune responses. Furthermore, they are consistent with the recent observation that HLA-DR positive keratinocytes may modulate cutaneous immunological reactions by inducing T-cell unresponsiveness.

Topics: Adult; Antigens, Differentiation; CD36 Antigens; Dermatitis, Atopic; Epidermis; Humans; Keratins; Lichen Planus; Lupus Erythematosus, Discoid; Lymphoma; Monocytes; Psoriasis; Skin Diseases; Skin Neoplasms

Psoriasis is a common papulosquamous skin disease. The histopathology is characterized by epidermal hyperplasia and inflammation. Recent studies suggest that keratinocyte proliferation and inflammation in psoriasis are manifestations of the same underlying pathological process. Interleukin 6 (IL-6), a cytokine that is a major mediator of the host response to tissue injury and infection, is produced by both keratinocytes and leukocytes in culture. IL-6 expression was studied in psoriatic plaques by immunoperoxidase staining with two different polyclonal anti-recombinant IL-6 antisera and by in situ nucleic acid hybridization with IL-6 cRNA probes. Epidermal and dermal cells in active psoriatic plaques from 35 psoriasis patients stained heavily for IL-6 as compared with nonlesional skin and with plaques after treatment with antimetabolic and antiinflammatory agents. Absorption of the anti-recombinant IL-6 antisera with purified fibroblast-derived IL-6 or with recombinant IL-6, but not bovine serum albumin, removed the immunostaining. Increased levels of IL-6 were detected in the plasma of patients with active psoriasis (mean 3 ng/ml) by using two different bioassays. IL-6 production by proliferating keratinocytes was suggested by IL-6-specific immunostaining in cultured normal and psoriatic keratinocytes and by the detection of mRNA specific for IL-6 in psoriatic epidermis by in situ hybridization. IL-6 stimulated the proliferation of cultured, normal human keratinocytes as assessed by two different assays. Thus, IL-6 could directly contribute to the epidermal hyperplasia seen in psoriatic epithelium as well as affect the function of dermal inflammatory cells.

Topics: Cell Division; Cells, Cultured; Epidermal Cells; Epidermis; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Interleukin-6; Interleukins; Keratins; Nucleic Acid Hybridization; Psoriasis; Recombinant Proteins; RNA, Messenger; Skin; Transcription, Genetic

Assessment of antikeratin antibodies (AKAs), using an indirect immunofluorescence technique with rat esophageal keratin as antigen, was performed in 14 patients with arthropathy and palmoplantar pustular eruptions, six of whom also had psoriasis vulgaris. Twelve patients had seronegative spondyloarthropathy. They were all AKA negative. Two patients had classical seropositive-erosive rheumatoid arthritis (RA), and were both AKA positive. This suggests that AKA is not related to the arthropathy associated with pustulosis palmoplantaris or psoriasis, or to the presence of pustular eruptions on the palms and soles. The finding of AKA in RA is in keeping with previous findings.

Topics: Aged; Autoantibodies; Dermatitis; Female; Humans; Immunoglobulin G; Joint Diseases; Keratins; Male; Middle Aged; Psoriasis

Keratinocytes from involved psoriatic plaques (PP), uninvolved, clinically symptomless skin of psoriatic patients (PN) and normal healthy skin (NN) have been cultured in a low calcium serum free system for multiple passages. In this way, the keratinocytes were removed from microenvironmental factors present in the skin. While the basal rate of proliferation of the PP, PN and NN keratinocytes was not different, the PP cells produced more transforming growth factor-alpha (TGF-alpha) than NN cells, and the antiproliferative response of PP cells to gamma interferon (IFN-gamma), a product of activated T lymphocytes, was reduced. We studied IFN-gamma because it can inhibit the proliferation of NN keratinocytes, induce their differentiation and the appearance of two immunoregulatory cell surface molecules, HLA-DR and intercellular adherence molecule-I (ICAM-I), and because in another epithelial cell system, epidermal growth factor (EGF) modulates IFN-gamma activity. The mean antiproliferative effects of IFN-gamma at 50,200, and 500 U/ml for the PP group (n = 10) was less compared to the NN group (n = 11); P less than 0.001, while the PN group (n = 5) had a less dramatic, but statistically significant, reduction in growth inhibition by IFN-gamma only at 200 and 500 U/ml compared to NN cells; P less than 0.05 and P less than 0.01, respectively. The amount of TGF-alpha produced and secreted by PP keratinocytes from five different individuals was significantly greater than by NN keratinocyte cultures. In addition, IFN-gamma induced TGF-alpha to a lesser extent in PP keratinocytes compared to NN keratinocyte cultures. Keratinocytes isolated from atopic dermatitis and Sézary syndrome patients were similar to NN keratinocytes. In contrast to its differential effects on TGF-alpha production and proliferation, IFN-gamma induced similar amounts of HLA-DR and ICAM-I on PP, PN and NN keratinocytes. Thus, for the PP keratinocytes, there was a dissociation between the antiproliferative and immunomodulatory effects of IFN-gamma. These results support our previous hypothesis that the hyperproliferation and altered differentiation of keratinocytes in psoriatic plaques is linked to an altered responsiveness of the keratinocytes to IFN-gamma. Moreover, these results provide an in vitro correlate of our in vivo observation of increased TGF-alpha levels in psoriatic plaques. A new pathophysiological model to understand psoriasis is proposed which integrates these observations involv

Topics: Adult; Cell Division; Cells, Cultured; Epidermal Cells; Epidermis; Female; Humans; In Vitro Techniques; Interferon-gamma; Keratins; Male; Middle Aged; Psoriasis; Recombinant Proteins; Transforming Growth Factors

We have used monoclonal antibodies to study the expression of calgranulins by keratinocytes in inflammatory dermatoses. Calgranulins are intracellular calcium binding proteins which have inflammatory cytokine activity and are composed of at least two different chains, calgranulin A and B. Antibody CF 145 and CF 557 identify calgranulin A and B, respectively. MAC 387 recognizes a molecule probably containing both calgranulins. Keratinocytes in normal skin did not contain these molecules. The keratinocytes in 52 cases of different inflammatory dermatoses showed expression of both calgranulin chains in lesional but not in non-lesional skin. Keratinocytes in inflammatory dermatoses therefore express an intracellular calcium binding protein which has cytokine activity.

Topics: Antibodies, Monoclonal; Calcium-Binding Proteins; Calgranulin A; Calgranulin B; Dermatitis, Contact; Epidermis; Graft vs Host Disease; Humans; Keratins; Psoriasis; Skin Diseases, Vesiculobullous

Keratin-type intermediate filament proteins show characteristic expression in normal and pathologic epidermis. Some keratins are restricted to the basal cell layers, and others occur exclusively in the suprabasal compartment. SDS-gel-electrophoresis and immunohistochemistry are generally used for the assessment of keratin profiles and their localizations. In the present investigation, flow cytometric analysis of four different monoclonal antibodies (MAb) against intermediate filament-type proteins, in addition to measurement of relative DNA content, was performed on cell suspensions derived from lesional and clinically uninvolved skin of psoriatic patients and from skin of healthy controls. MAb Ks8.12, reacting with keratins 13 and 16, was used as a marker for hyperproliferation. Pab601 recognizes the basal cell layer(s) of human epidermis. Keratin 10 expression as a marker of keratinization was quantified with RKSE60 and the anti-vimentin MAb MVI was used as a marker for non-keratinocytes. Psoriatic skin showed significantly reduced numbers of RKSE60-positive cells and MVI-positive cells compared with normal skin. In contrast to normal skin and uninvolved skin of psoriatic patients in which only a minority of the cells were Ks8.12 positive, up to 60% of the cell population in psoriatic lesions bound with MAb. Simultaneous measurement of relative DNA content and MAb binding showed that Pab601 binding was associated with cells in S-phase and G2M-phase of the cell cycle, whereas RKSE60 and Ks8.12 binding were associated with diploid cells. Multiparameter flow cytometry allows quantitative population analysis that could lead to a better understanding of the complex mechanisms of epidermal growth control under normal and pathologic conditions.

Topics: Adult; Antibodies, Monoclonal; DNA; Epidermis; Flow Cytometry; Fluorescent Antibody Technique; Humans; Intermediate Filament Proteins; Interphase; Keratins; Middle Aged; Psoriasis

Recently the treatment of psoriasis with vitamin D3, its metabolites or analogues, has been reported to be clinically effective and free from side effects. We have quantified the changes in the levels of epidermal lesional keratins in 15 psoriatic patients receiving oral 1 alpha(OH)D3 treatment. Lesions were sampled before treatment commenced and at monthly intervals for 4-6 months. Clinical resolution occurred in 7 patients within this time; 3 other patients showed incomplete lesion resolution and the remaining 5 patients showed a complete lack of response within the treatment period. Lesion resolution, as judged by clinical criteria, was accompanied by significant changes in the levels of three of the keratin polypeptides and smaller changes in others. Keratin 2 increased to levels greater than those in normal epidermis, while keratins 16 and 18 decreased to normal levels. Changes in the levels of keratins 1 and 5 were small and those of keratins 7, 10 and 14 minimal. These changes were compared with values found during lesion resolution with other therapies used in psoriasis, i.e. topical dithranol, PUVA, oral etretinate and hydroxyurea and were highly reminiscent of those observed during PUVA therapy but contrasted with those during etretinate treatment. The decrease in level of keratin 16, a hyperproliferation marker, suggests that 1 alpha(OH)D3 inhibits keratinocyte proliferation, but at the same time the overproduction of keratin 2, a major keratin of the granular cells, indicates that there is an increase in the number of cells in the later stages of differentiation.

Topics: Anthralin; Calcium; Cell Differentiation; Cell Division; Densitometry; Electrophoresis, Polyacrylamide Gel; Etretinate; Female; Humans; Hydroxycholecalciferols; Hydroxyurea; Immunoblotting; Keratins; Male; Phosphates; Psoriasis; PUVA Therapy; Skin

Quantitative changes in the levels of keratin polypeptides extracted from keratotome shavings from psoriatic epidermis were measured by using one-dimensional SDS-PAGE, followed by scanning densitometry. Values obtained were compared with results for non-lesional epidermis and from epidermis from normal individuals. Patients on four different treatment regimens were investigated by repeated sampling over 3-4 months starting before therapy commenced. The levels of four keratins changed significantly: keratins I (70 kd) and 2 (66 kd) tended to rise to normal levels, while keratins 16 (50 kd) and 18 (44 kd) fell to normal levels. There were differential effects as well as differences in the rates of normalization depending upon the treatment regimen. The most rapid normalization of the levels of all four keratins was observed with topical dithranol (anthralin) treatment (five patients) with plaque resolution and keratin level normalization after 7-9 weeks. Oral hydroxyurea (three patients) had similar effects, but over a longer time scale (20 weeks). In contrast, oral etretinate (four patients) caused a normalization of all except keratin 2 (66 kd) over a period of 20-28 weeks, and keratin I (70 kd) levels tended to 'overshoot' the normal level. PUVA (five patients) caused rapid normalization (in 9-12 weeks) of keratins 2 (66 kd) and 18 (44 kd), but had much weaker effects on keratins I (70 kd) and 10 (57 kd). These results suggest that resolution of lesions as judged by clinical criteria can occur without normalization of the keratin electrophoretic profile. Possibly the most reliable marker of clinical resolution was the reduction in keratin 16 (50 kd), since treatment effects on the differentiation of keratins I (70 kd) and 2 (66 kd) were different.

Topics: Anthralin; Densitometry; Electrophoresis; Epidermis; Etretinate; Female; Gene Expression; Humans; Hydroxyurea; Immunoblotting; Keratins; Male; Molecular Weight; Psoriasis; PUVA Therapy

It is now well established that epidermis, like many other tissues, contains a phospholipase A2 that is responsible for the initiation of the arachidonic acid cascade. Here we report that human epidermis also contains a second, quite distinct enzyme of the phospholipase A2 group, which is unique in its extreme activity against phospholipids in true solution. It also differs from the classic cutaneous enzyme in that (a) its activity is not reduced by pretreatment of the skin with corticosteroids in vivo nor by treatment of the epidermal homogenate with alkaline phosphatase in vitro, and (b) its activity is reduced, rather than increased, in the lesions of inflammatory diseases such as psoriasis. The enzyme seems to occur mainly in fully differentiated keratinocytes, its level being low in the basal cell layer of epidermis and in keratinocytes cultured in vitro. On the basis of these observations, we suggest that this new phospholipase A2 is responsible for the degradation of phospholipids that accompanies the terminal keratinization process.

Topics: Alkaline Phosphatase; Cells, Cultured; Clobetasol; Eczema; Epidermis; Humans; Keratins; Lichen Planus; Phospholipases; Phospholipases A; Phospholipases A2; Psoriasis; Skin Diseases; Trypsin

Psoriatic epidermis is characterized by increased DNA synthesis and disturbed differentiation. Even though these processes are closely associated, most investigations do not give insight into temporal/spatial relationships between both events. We previously developed a double labeling method for the simultaneous demonstration of the germinative and differentiated epidermal compartments in normal human skin by using tritium-labeled thymidine ([3H] Thd) incorporation and immunoperoxidase staining of 67 kD keratin polypeptides. In this paper we report the results of combined evaluation of these compartments in stable plaques of psoriasis. Scanning of skin sections with an automatic image analyzer allows objective quantification of areas of total epidermis, 67 kD+ differentiated epidermis and numbers of [3H] Thdr+ nuclei. Our data indicate that the 67 kD- undifferentiated psoriatic epidermis is expanded. Increased numbers of [3H] Thd+ basal and suprabasal psoriatic keratinocytes are present and most of them (97.9%) pertain to the 67 kD- compartment. Keratin identification in scales taken from the same sites showed a variable but distinct decrease of 67 kD keratin polypeptides. Hence, the hyperplastic epidermis of stable plaques of psoriasis is characterized by the presence of increased numbers of [3H] Thd+ cells, which primarily belong to the undifferentiated (67 kD-) basal and suprabasal compartments, especially in the lowermost parts of the elongated interpapillary rete ridges. These changes are associated with a relative decrease of synthesis of 67 kD polypeptides and the presence in the scales of keratins that confer a characteristic hyperproliferative epidermal keratin pattern to the psoriatic plaque.

Topics: Cell Differentiation; DNA; Epidermis; Humans; Image Processing, Computer-Assisted; Keratins; Psoriasis

Epidermis in cryostat sections of skin biopsy specimens from patients with psoriasis and from healthy individuals bound bovine erythrocytes (E) sensitized with rabbit IgG antibodies (A)(EA). No binding occurred using E or E sensitized with IgM or F(ab')2 fragments of IgG. The binding of EA was inhibited by human IgG and by Fc fragments of IgG, whereas human IgA, IgM, albumin, and F(ab)2 fragments of IgG did not inhibit the binding, indicating the presence of receptors for the Fc part of IgG (FcR). EA bound mainly to stratum spinosum and most strongly above FcR-positive cell infiltrates in dermal papillae. The binding of EA to sections from patients with active psoriasis was stronger than to sections from patients with stationary psoriasis vulgaris. Sections of unaffected skin from patients with psoriasis and healthy individuals also bound EA, but the binding was weaker than to sections of psoriatic lesions. The receptors were sensitive to periodic acid, formaldehyde, and heat. Using immune complexes of horseradish peroxidase (HRP) and rabbit IgG antibodies to HRP, the receptors were localized to the outer aspect of the keratinocytes and to the inflammatory cells in the microabscesses. The strongest binding occurred in the same areas which adhered EA most strongly. FcR on dendritic epidermal cells could not be demonstrated in situ. A monoclonal antibody against FcR also stained the outer aspect of most keratinocytes throughout the epidermis. FcR on keratinocytes support the assumption that these cells contribute to immune reactions in the skin.

Topics: Antigen-Antibody Complex; Epidermis; Erythrocytes; Horseradish Peroxidase; Humans; Immunoglobulin G; Keratins; Middle Aged; Psoriasis; Receptors, Fc; Receptors, IgG

Topics: Antibodies, Monoclonal; Epidermal Cells; Epidermis; Fluorescent Antibody Technique; Humans; Keratins; Psoriasis; Stem Cells

The formation of a cornified envelope (CE) is a major event in the terminal differentiation of epidermal cells. Nomarski contrast microscopy of the envelopes purified from different sources reveals the existence of two major, but morphologically distinct classes: the very irregularily shaped fragile type CEf, and the polygonal rigid type CEr. Human keratinocytes in submerged culture are only able to produce type CEf. Specimens from healthy human epidermis contain largely type CEr. Psoriatic scales from different patients show both types in varying proportions. Tape stripping of normal epidermis reveals that type CEf is present in the lowermost layers of the stratum corneum and type CEr is present in the upper layers, indicating that the two types represent a different stage of maturation. Cyanogen bromide peptide mapping of electrophoretically purified envelopes reveals striking differences between cultured keratinocytes, normal epidermis, and psoriatic scales but also slight interindividual variations. This variability supports the view that the molecular CE composition is not strictly determined. On the other hand, no difference could be detected in the peptide maps of CEf and CEr obtained after tape stripping from the same healthy volunteer indicating that CE maturation within the stratum corneum does not involve the provision of qualitatively new proteins.

Topics: Cell Differentiation; Cells, Cultured; Cyanogen Bromide; Electrophoresis, Polyacrylamide Gel; Epidermal Cells; Epidermis; Humans; Keratins; Peptides; Psoriasis; Reference Values

Cyclosporin A, which is a specific immunosuppressive compound, has recently been demonstrated to be of significant benefit in the treatment of psoriasis. Because hyperplasia is a major feature of psoriasis, we have investigated whether this drug acts directly to inhibit keratinocyte growth. We have determined the concentration range of cyclosporin in the epidermis of psoriatic patients undergoing cyclosporin therapy and the effect of this concentration range on the growth of cultured keratinocytes. After 7 days of treatment, psoriatic involved epidermis contained 1.1 +/- 0.3 ng cyclosporin/micrograms DNA. Based on tissue wet weight this is approximately 2.8 micrograms cyclosporin/ml. This value was 10 times that of trough blood samples. On day 3 of treatment, involved epidermis contained 7 times more cyclosporin than scale, while on day 7 this ratio was reduced to 2. This suggests that cyclosporin was initially associated with the lower layers of the epidermis and distributed upward with time. Exposure of adult human keratinocytes, cultured on collagen in the presence of human serum, to cyclosporin (0.1-30 micrograms/ml, 0.4-13-fold higher than lesional cyclosporin) for 2-6 d had no effect on the rate of incorporation of thymidine into DNA. Cyclosporin (1-30 micrograms/ml) also had not effect on the reinitiation of DNA synthesis of quiescent cells subsequent to the reintroduction of serum. In contrast, cyclosporin (1-10 micrograms/ml) inhibited growth of keratinocytes cultured on plastic culture dishes in serum free media (MCDB 153). For a given dose of cyclosporin, cell associated drug was 2-3 times greater in serum free compared to serum containing media. This may contribute in part to the sensitivity of keratinocytes in serum free media to growth inhibition by cyclosporin. These data demonstrate that human epidermis contains a high concentration of cyclosporin after oral administration, and that, under certain conditions, concentrations of cyclosporin within the range found in vivo can inhibit growth of cultured keratinocytes. Because cyclosporin regulates lymphocyte function in vivo and in vitro, the demonstrated antiproliferative effects of cyclosporin on psoriatic keratinocytes in vivo may be due to inhibition of a mononuclear leukocyte-derived keratinocyte growth factor(s) in combination with direct inhibition of keratinocyte growth.

Topics: Blood Physiological Phenomena; Calcium; Cells, Cultured; Culture Media; Cyclosporins; DNA; Dose-Response Relationship, Drug; Epidermal Cells; Epidermis; Humans; Keratins; Psoriasis

The ultrastructure of human affected and unaffected psoriatic epidermis was studied in skin biopsies from 5 patients and 3 normal controls. Transmission electron microscopic investigations revealed abnormalities in all cell layers of the affected epidermis. Common to psoriatic keratinocytes from affected epidermis was the reduction of tonofilaments. The essential ultrastructural changes were located in the stratum granulosum and stratum corneum. Thus, absence of the fusion between the keratohyalin granules and the tonofilaments was found in stratum granulosum. The keratinocytes of the stratum corneum showed a large accumulation of ribosomes and vesicles resembling lipid vesicles.

Topics: Actin Cytoskeleton; Adult; Epidermis; Humans; Keratins; Microscopy, Electron; Middle Aged; Psoriasis

We report here the isolation and characterization of three antisera, each of which is specific for a single keratin from one of the three different pairs (K1/K10, K14/K5, K16/K6) that are differentially expressed in normal human epidermis and in epidermal diseases of hyperproliferation. We have used these antisera in conjunction with monospecific cRNA probes for epidermal keratin mRNAs to investigate pathways of differentiation in human epidermis and epidermal diseases in vivo and in epidermal cells cultured from normal skin and from squamous cell carcinomas in vitro. Specifically, our results suggest that: (a) the basal-specific keratin mRNAs are down-regulated upon commitment to terminal differentiation, but their encoded proteins are stable, and can be detected throughout the spinous layers; (b) the hyperproliferation-associated keratin mRNAs are expressed at a low level throughout normal epidermis when their encoded proteins are not expressed, but are synthesized at high levels in the suprabasal layers of hyperproliferating epidermis, coincident with the induced expression of the hyperproliferation-associated keratins in these cells; and (c) concomitantly with the induction of the hyperproliferation-associated keratins in the suprabasal layers of the epidermis is the down-regulation of the expression of the terminal differentiation-specific keratins. These data have important implications for our understanding of normal epidermal differentiation and the deviations from this process in the course of epidermal diseases of hyperproliferation.

Topics: Amino Acid Sequence; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cell Transformation, Neoplastic; Cells, Cultured; Epidermal Cells; Epidermis; Humans; Immune Sera; Immunoassay; Immunohistochemistry; Keratins; Molecular Sequence Data; Nucleic Acid Hybridization; Psoriasis; RNA, Messenger; Skin Neoplasms; Tumor Cells, Cultured

We investigated the effects of 1,25-dihydroxyvitamin D3 (1,25-(OH)2-D3) on cultured fibroblasts and keratinocytes from patients with psoriasis and treated 17 patients with psoriasis with orally or topically administered 1,25-(OH)2-D3. Cultured fibroblasts from three of five patients showed a normal response to the antiproliferative activity of a physiologic dose of 1,25-(OH)2-D3, whereas fibroblasts from the other two had a partial resistance to the drug. Cultured keratinocytes from two patients with psoriasis possessed nuclear receptors for 1,25-(OH)2-D3 and the drug caused a dose-dependent inhibition of proliferation and induction of terminal differentiation of these cells similar to effects in normal cultured keratinocytes. Ten of 14 patients with moderate to severe psoriasis who received oral 1,25-(OH)2-D3 showed significant clearing of their hyperkeratotic plaques. Three patients had complete clearing that was sustained with maintenance therapy, but four patients received little or no benefit from the therapy. By the administration of 1,25-(OH)2-D3 as a single oral dose at bedtime, larger doses of the drug could be tolerated without evidence of hypercalciuria or hypercalcemia. Three patients who received topical 1,25-(OH)2-D3 showed a rapid response with complete clearing after 6 weeks of therapy. Therefore, these preliminary findings suggest that orally or topically administered 1,25-(OH)2-D3 may be a safe and effective alternative therapy for the treatment of psoriasis.

Topics: Administration, Cutaneous; Administration, Oral; Adult; Biopsy; Calcitriol; Calcium; Double-Blind Method; Epidermis; Female; Fibroblasts; Humans; In Vitro Techniques; Keratins; Male; Middle Aged; Psoriasis; Recurrence

The pathologic features of psoriatic plaques are inflammation and increased epidermal turnover. IP-10, a cytokine the expression of which is induced by gamma-interferon, is a member of a family of soluble mediators with inflammatory and growth-promoting activities. IP-10 protein was detected in keratinocytes and the dermal infiltrate from active psoriatic plaques using an affinity-purified rabbit anti-IP-10 antibody in immunoperoxidase studies. Successful treatment of active plaques decreased IP-10 expression in plaques. These results were corroborated by Northern blot analysis with an IP-10 cDNA probe. We have previously detected activated T cells and HLA-DR keratinocytes in active psoriatic plaques. Since IP-10 is detected in delayed cellular immune responses, the present study further points to the role of ongoing cellular immune responses in the pathogenesis of psoriasis.

Topics: Blotting, Northern; Chemokine CXCL10; Chemokines, CXC; Epidermis; Humans; Immunoenzyme Techniques; Interferon-gamma; Keratins; Proteins; Psoriasis; RNA, Messenger

Recent studies suggest that the immune system is involved in the pathogenesis of psoriasis. We studied the expression and distribution of immunocompetent cells and of some chosen differentiation antigens of keratinocytes at various stages of lesion development, using indirect and amplified immunofluorescence, and avidin-biotin-peroxidase method. Serial cryostat sections were collected so as to allow comparative studies of adjacent parts of each biopsy sample with various immunocytochemical markers. Our results indicate that focal intra-epidermal infiltration of otherwise unaltered epidermis with lymphocytes, mostly of the T-helper phenotype, was the first perceptible change occurring in patients with eruptive psoriasis. Modification of the Langerhans' cell staining was observed in these initial subclinical lesions. A significant reduction of the cell frequency was noted in psoriatic papules and plaques. Changes in the epidermal antigen expression could be observed in the developed lesions only. The simultaneous appearance of histologic signs of psoriasis and the modification of keratinocyte antigens indicates that both events are related to the epidermis hyperproliferation, possibly induced by focal inflammatory reaction.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antigens, Differentiation; Dendritic Cells; Epidermis; Female; HLA-DR Antigens; Humans; Keratins; Lymphocytes; Male; Middle Aged; Psoriasis

Topics: Epidermis; Humans; Keratins; Microscopy, Electron; Mitochondria; Psoriasis

Human cell hybrids derived from fusions between carcinoma (HeLa) and normal epidermal keratinocytes can be used as immunogens to generate monoclonal antibodies against keratinocyte differentiation-specific antigens. We examined the immunofluorescent staining patterns of these monoclonals in normal human and psoriatic (lesional and uninvolved) skin tissue sections. The immunofluorescent staining patterns indicate that the monoclonal antibodies are recognizing a number of different keratinocyte antigens. Three monoclonals (HLK7, HLK3, and HLK20) displayed different immunofluorescent reactivity in these tissues. These new monoclonal antibodies will be useful tools to study the differentiation abnormalities in psoriasis and may serve as a marker of the psoriatic diathesis.

Topics: Antibodies, Monoclonal; Antigens, Differentiation; Biopsy; Cells, Cultured; Epidermal Cells; Fluorescent Antibody Technique; Humans; Keratins; Psoriasis

Topics: Epidermis; Humans; Immunohistochemistry; Keratins; Psoriasis

We have investigated the immunoperoxidase staining pattern in the epidermis and dermal infiltrates of highly inflamed portions of psoriatic lesions, selecting for biopsy early pinpoint lesions or margins of active plaque lesions. We found positive intercellular staining for HLA-DR antigens in localized areas of the epidermis in about half of the patients tested. In contrast, OKT6 antigen was found only on the dendritic cells in the epidermis and dermis in all cases. These findings support the hypothesis that an active cellular immune reaction involving the epidermis, possibly associated with the expression of HLA-DR antigens on keratinocytes, occurs in the highly inflamed areas of psoriatic lesions, particularly in early pinpoint lesions or at the edges of spreading plaque lesions.

Topics: Dermatitis; Epidermal Cells; Epidermis; HLA-D Antigens; HLA-DR Antigens; Humans; Immunoenzyme Techniques; Keratins; Psoriasis

Protein extracts from normal human epidermis reassemble in vitro into 8-10 nm diameter filaments characteristic of intermediate filaments, whereas extracts from psoriatic epidermal scales reassemble, under identical conditions, into a variety of paracrystalline bundles. Optical diffraction and image analysis of these paracrystalline bundles reveal an axial repeat of 16.5 nm, which subdivides into three bands of 5.5 nm, and a lateral spacing of 5.1 nm. This information, together with available sequence studies of intermediate filaments and biochemical data, suggests that the subunit of psoriatic keratin is made up essentially from the coiled-coil alpha-helical rod domain of the normal keratin subunits, whereas the random coil domains are missing or greatly reduced in size.

Topics: Cytoskeleton; Humans; Image Processing, Computer-Assisted; Intermediate Filaments; Keratins; Microscopy, Electron; Psoriasis

We have measured the RNA and DNA content and examined cell surface characteristics of human epidermal cells derived from normal skin, and lesional and nonlesional areas of psoriatic skin prior to and following treatment on a modified Goeckerman protocol. Our results show that cells from active psoriatic lesions contain greater numbers of basal keratinocytes when compared with either nonlesional skin from the same patients or skin from healthy volunteers and individuals with other inflammatory skin lesions. Follow-up measurements 2-3 weeks after the initiation of therapy showed that the numbers of basal keratinocytes in resolving psoriatic lesions had decreased and approached normal levels. Multiparameter RNA/DNA flow cytometric analysis on parallel samples from the same psoriasis patients revealed an increased growth fraction and proportion of cycling cells in both the nonlesional and lesional skin compared with controls. Furthermore, the cellular RNA content was elevated in lesional psoriatic skin when compared with either nonlesional or normal skin. Flow cytometric examination of nonlesional and lesional epidermal cells obtained 2-3 weeks after the commencement of therapy revealed that the growth fraction and mean RNA content of the keratinocytes from resolving psoriatic plaques decreased in response to therapy. In contrast, the proportion of keratinocytes within the S + G2 + M phases of the cell cycle remained elevated. These data indicate that "uninvolved" psoriatic skin exhibits characteristics more closely resembling lesional psoriatic skin than normal skin. The results further suggest that quantitation of cellular RNA content and basal cell number might be sensitive indicators of early treatment response in psoriasis.

Topics: Adult; Aged; Cell Count; Cell Membrane; DNA; Epidermis; Female; Flow Cytometry; Humans; Keratins; Male; Middle Aged; Psoriasis; RNA; Skin

Basal keratinocyte herniations (BKH) have been used as markers of psoriatic activity. Abnormal multipolypoid forms herniating through large gaps in the basal lamina have been found to characterize biopsies from psoriatic patients with concomitant alpha 1-antitrypsin deficiency, and appear to be a marker of excessive proteolytic activity. The finding of similar multipolypoid BKH in patients with generalized pustular psoriasis of the von Zumbusch variety (but not in patients with psoriasis vulgaris), in the absence of concomitant alpha 1-antitrypsin deficiency, would support the concept of the presence of large amounts of proteolytic enzymes in the dermis of patients with this syndrome. The large proportion of BKH directly associated with dermal neutrophils, and the presence of clusters of high-density BKH overlying collections of dermal neutrophils, suggests that neutrophilic proteases are largely responsible for BKH formation in patients with this syndrome.

Topics: Adult; alpha 1-Antitrypsin; Epidermis; Female; Humans; Keratins; Male; Microscopy, Electron; Middle Aged; Neutrophils; Phenotype; Psoriasis; Skin

Topics: Animals; Cell Division; Cells, Cultured; Fluorescent Antibody Technique; Histocytochemistry; Humans; Immunoenzyme Techniques; Keratins; Ornithine Decarboxylase; Psoriasis; Skin; Skin Neoplasms; Ultraviolet Rays

Topics: Anthralin; Epidermis; Humans; Keratins; Microscopy, Electron; Mitochondria; Psoriasis

Immunocytochemical studies with a monoclonal anti-HLA-DR antibody were performed on skin sections and keratinocyte (KTC) suspensions obtained from suction blisters of active psoriatic plaques. HLA-DR+ KTCs were found in the plaques of 23 of 38 patients with active psoriasis. Of these 23, 16 had clinical findings typical of psoriatic arthritis (PA); none of the 15 patients who lacked HLA-DR+ KTCs had PA. Although KTC HLA-DR expression was more prevalent in patients with severe skin disease, 7 of the 23 patients with HLA-DR+ KTCs in active psoriatic plaques had mild skin disease; 4 of these 7 had PA. Nail pitting or duration of skin disease did not account for increased incidence of PA in patients with HLA-DR+ KTCs. All psoriasis patients with arthritis received nonsteroidal antiinflammatory drug therapy; 14 received additional therapy directed primarily to the cutaneous manifestations of psoriasis. Nine of these noted arthritis improvement with concurrent skin response; however, in 5 patients, arthritis activity increased, despite improvement of the cutaneous disease. Two other patients, treated with methotrexate, also had concurrent skin and joint improvement. These data suggest that psoriasis patients with HLA-DR+ KTCs are at increased risk for the development of associated arthritis.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Arthritis; Epidermis; Histocytochemistry; HLA-D Antigens; HLA-DR Antigens; Humans; Immunochemistry; Keratins; Psoriasis

Psoriatic and control human hair follicle keratinocytes were cultured on bovine eye lens capsules in Epicult dishes for a period of 5-6 weeks and examined using light microscopy. The following morphological differences between cultures were observed: 1. The lower cell layers contained predominantly flattened cells in psoriatic cultures instead of roundish in control cultures. 2. The differentiation pattern was irregular in psoriatic cultures instead of regular in control cultures. 3. The differentiated zone of psoriatic cultures was more compact and thicker in comparison to normal cultures. These differences might allow discrimination between normal and psoriatic cultures.

Topics: Cells, Cultured; Epidermal Cells; Hair; Humans; Keratins; Psoriasis

Membrane-associated thioredoxin reductase (TR) activity has been measured in 3-mm punch biopsy specimens from psoriatic and uninvolved skin in eight patients with chronic plaque-type psoriasis vulgaris. The mean specific activity for this free radical reducing enzyme in psoriatic vs uninvolved skin was 28.5 +/- 8.0 U vs 16.8 +/- 4.25 U. Because TR contains two reactive thiolate groups at its active site, this enzyme reacts with anthralin to form a covalent anthralin-TR complex causing irreversible enzyme deactivation. This mode of action for anthralin was confirmed by using pure TR from Escherichia coli. Keratinocyte cell cultures, grown from normal and psoriatic skin of one donor, revealed 24% and 42% inhibition of cell surface TR activity, respectively, in the presence of 2 X 10(-5)M anthralin. Time-dependent topical application of anthralin on guinea pig skin gave 70% inhibition of TR with concentrations of 0.25% to 1.0% after 24 hours in open and occlusive applications of the drug. Short contact with 1% anthralin showed 70% inhibition after 120 minutes.

Topics: Adult; Animals; Anthralin; Cells, Cultured; Epidermal Cells; Female; Guinea Pigs; Humans; Keratins; Male; NADH, NADPH Oxidoreductases; Psoriasis; Skin; Thioredoxin-Disulfide Reductase; Time Factors

The surface structure of 11-12 days old cultures from biopsies of normal skin and uninvolved psoriatic skin was investigated by scanning electron microscopy. The keratinocytes formed flat, thin, and well-organized layers of elongated, tightly apposed cells. Newly-formed cells with few and short microplicae and microvilli were seen at the colony periphery. On the maturing cells towards the colony center, the number and size of microplicae or microvilli increased gradually. No differences were found between the structures on normal keratinocytes and keratinocytes from uninvolved psoriatic skin, but in keratinocytes from involved psoriatic skin glomerulus-like rolled microplicae occasionally were found.

Topics: Adult; Cells, Cultured; Epidermis; Humans; Keratins; Microvilli; Middle Aged; Psoriasis

Topics: Drug Evaluation; Etretinate; Humans; Keratins; Lichen Planus; Psoriasis; PUVA Therapy; Retinoids; Skin; Skin Diseases; Time Factors

Topics: Animals; Epidermis; Humans; Keratins; Mice; Psoriasis; T-Lymphocytes

Topics: Adolescent; Adult; Aged; Autoantibodies; Child; Electrophoresis; Female; Humans; Intermediate Filament Proteins; Keratins; Male; Middle Aged; Psoriasis

Topics: Adolescent; Adult; Aged; Autoantibodies; Child; Enzyme-Linked Immunosorbent Assay; Female; Humans; Intermediate Filament Proteins; Keratins; Male; Middle Aged; Psoriasis; Skin

Topics: Adrenergic beta-Antagonists; Aged; Epidermis; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Keratins; Middle Aged; Psoriasis

Three keratin antibodies (RKSE 60, Clone 77, and a rabbit polyclonal) and 2 vimentin antibodies (Vim ab and a rabbit polyclonal) were investigated using frozen sections of normal and psoriatic skin. Of these, the monoclonals RKSE 60 and Vim ab were selected for quantitative population analysis of healthy epidermis, psoriatic uninvolved epidermis, and psoriatic lesions. Suspensions of isolated cells were prepared from biopsy specimens by trypsinization, and stained with RKSE 60 or Vim ab using an indirect immunofluorescence assay. Our results showed an increase in the germinative fraction from the normal value of 30% to almost 50% in the psoriatic lesion; in absolute terms this corresponds to a 6-fold increase in the size of the germinative compartment. More interesting, the germinative psoriatic uninvolved epidermis (38%) was also significantly higher than normal. The percentage of vimentin-positive cells (Langerhans cells and melanocytes) was nearly double that of normal in both the lesion and the uninvolved psoriatic epidermis. We conclude that, in contrast to statements frequently encountered in the literature, the "uninvolved" skin of the patient is morphologically and functionally different from that of the healthy individual.

Topics: Adult; Antibodies, Monoclonal; Cell Differentiation; Epidermal Cells; Fluorescent Antibody Technique; Frozen Sections; Humans; Keratins; Middle Aged; Psoriasis; Vimentin

Using transmission electron microscopy, we studied, quantitatively, basal keratinocyte herniations (BKH) in relation to the other basement membrane zone changes in psoriatic lesions of varying clinical activity, and in psoriasiform skin diseases. BKH appears to correlate with disease activity. They do not occur passively as a result of the formation of gaps in the basal lamina. BKH in active psoriasis are associated with electron-lucent areas suggestive of proteolytic enzyme release. Their apparent association with Langerhans cells, neutrophils, macrophages, and endothelial cells may point to these cells as the source of proteolytic enzymes in psoriasis. BKH may prove to be a useful marker for clinical psoriasis.

Topics: Basement Membrane; Endothelium; Epidermal Cells; Epidermis; Female; Humans; Keratins; Langerhans Cells; Macrophages; Male; Microscopy, Electron; Neutrophils; Psoriasis

An antiserum prepared against a glycoprotein (GP37) extracted from the upper epidermal layers, was used to stain frozen sections of human oral mucosa, normal and abnormal skin by an indirect immunofluorescence technique. On normal human epidermis, this antiserum mainly reacted with the cytoplasm of granular cells, whereas on buccal mucosa the recognized antigen was observed as scattered dots limited to the upper epithelial layers. In epidermal diseases, alterations in the staining pattern were observed. In psoriasis, the labelling was markedly diminished; in contrast, in lichen planus it was intense and present on the 3-6 uppermost cellular layers. Basal cell epitheliomas were almost negative, except around horn cysts. In Bowen's disease dyskeratotic cells were strongly labelled. In squamous cell carcinomas a clear-cut staining was observed in squamous nests. On cultures, GP37 expression could be induced by growing epidermal cells in vitamin A-depleted medium. The biological significance of the observed staining patterns remains to be precised. Nevertheless, GP37 represents a sensitive marker of epidermal differentiation and may be useful in skin pathology and in in vitro studies.

Topics: Animals; Carcinoma, Squamous Cell; Epidermal Cells; Fluorescent Antibody Technique; Glycoproteins; Humans; In Vitro Techniques; Keratins; Lichen Planus; Macaca; Mice; Psoriasis; Rabbits; Skin Diseases; Skin Neoplasms

Topics: Cell Membrane; Epidermal Cells; Epidermis; Humans; Keratins; Lactoperoxidase; Membrane Proteins; Peptides; Psoriasis; Trypsin

Among several monoclonal antibodies (moABs) directed against human interleukin 2 (IL-2), the 15-2 moAB raised in our laboratory against unglycosylated recombinant IL-2 (produced in Escherichia coli) cross-reacted with a human skin epitope. This moAB gave a strong staining on the cell-surface membranes of keratinocytes from the granular layer of the epidermis. In addition, the 15-2 moAB stained 15% of epidermal cell suspensions obtained from suction blisters and reacted with cells from the spinous layer in parakeratosis and psoriasis, as well as with spinous epithelioma cells. Preincubation of the 15-2 moAB with pure human recombinant IL-2 abrogated skin binding, whereas a polyclonal antikeratin antiserum did not block 15-2 skin binding. Two other anti-IL-2 moABs, one directed against unglycosylated recombinant IL-2 (17-2 moAB) and one against glycosylated natural IL-2 (9B11 IE5 moAB), were unreactive on skin. Taken together, the data suggest that the 15-2 moAB binds to an epitope cross-reacting with, but different from, IL-2 which is located in the cell-surface membranes of granular layer cells. This cross-reactive epitope may provide a useful probe for the study of human epidermal cell differentiation.

Topics: Animals; Antibodies, Monoclonal; Binding Sites, Antibody; Epidermis; Humans; Interleukin-2; Keratins; Mice; Mice, Inbred BALB C; Parakeratosis; Psoriasis; Staining and Labeling

Topics: Carbon Radioisotopes; Epidermis; Glycine; Humans; Keratins; Kinetics; Protein Precursors; Psoriasis; Skin

Immunoperoxidase staining of skin sections and immunofluorescence analysis of keratinocyte suspensions obtained from suction blisters of psoriatic plaques were performed using an mAb, Josh 524.4.1, and Fab'2 fragments of a rabbit antiserum, both of which are directed against nonpolymorphic determinants of HLA-DR molecules. HLA-DR+ keratinocytes were present in plaques, but not normal-appearing skin, from a significant portion of patients with active psoriasis. Double-labelling immunofluorescence experiments with either the monoclonal or polyclonal anti-HLA-DR antibody, in conjunction with the mAb OKT6, which identifies DR+ Langerhans cells, demonstrated that HLA-DR molecules were present on OKT6- keratinocytes. The dermal infiltrate of psoriatic plaques contained T cells expressing the activation antigens, IL-2 receptor (Tac) and HLA-DR, as well as macrophages and OKT6+ cells. There was little difference in the characteristics of the dermal infiltrate between the lesions with or without HLA-DR+ keratinocytes. OKT6+ presumptive Langerhans cells were also found in the dermal infiltrates of patients with lichen planus, contact dermatitis, spongiotic dermatitis, erythema multiforme, basal and squamous cell carcinoma. Studies of keratinocyte suspensions showed that 7-84% of keratinocytes were HLA-DR+. Flow cytometry experiments showed that keratinocytes at all stages of differentiation were HLA-DR+. However, the stem cell-enriched population contained the highest proportion of HLA-DR+ cells. HLA-DR expression by keratinocytes correlated with disease activity. The expression was reversible with successful medical therapy. HLA-DR+ keratinocytes may activate T cells directly or may present an as yet unknown antigen to T cells. These studies provide further support for the hypothesis that immunological mechanisms play an important role in the pathogenesis of psoriasis.

Topics: Dermatitis; Epidermal Cells; Epidermis; HLA-D Antigens; HLA-DR Antigens; Humans; Interferon-gamma; Keratins; Langerhans Cells; Macrophages; Psoriasis; Skin; T-Lymphocytes

Certain arachidonic metabolites may play a pathogenic role in psoriasis. Platelets are rich sources of 12-hydroxy-eicosatetraenoic acid (12-HETE) and thromboxane A2, mediators of skin inflammation and platelet aggregation, respectively. We have studied untreated psoriatic patients without a history of diabetes mellitus and smoking. In psoriatics, platelet aggregation elicited by thrombin, ADP, and ristocetin was significantly enhanced as compared with healthy adult volunteers. Enhancement of platelet aggregation was detected in patients with both minimal and widespread disease. The formation of 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT), a cyclooxygenase product, and 12-HETE, a 12-lipoxygenase product, was increased in psoriatics with widespread disease but not in those with minimal disease. Formation of 12-HETE was stimulated to a higher degree (125%) than HHT (98%) in psoriasis (P less than 0.05). Addition of platelet-derived 12-HETE to cultured human epidermal keratinocytes resulted in a stimulation of the DNA synthesis (68% at 10(-7) M). These data suggest that platelet activation occurs in psoriasis, and that release of inflammatory and mitogenic compounds by activated platelets may play a role in the pathophysiology of psoriasis. Whether enhanced platelet aggregation in psoriasis is associated with occlusive vascular disease needs further investigation.

Topics: 12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; Blood Platelets; DNA; Epidermal Cells; Female; Humans; Hydroxyeicosatetraenoic Acids; In Vitro Techniques; Keratins; Male; Platelet Aggregation; Psoriasis; Stimulation, Chemical

Topics: Acitretin; Arachidonic Acid; Arachidonic Acids; Cell Line; Humans; Keratins; Psoriasis; Skin; Tretinoin; Ultraviolet Rays

Topics: Antibodies, Monoclonal; Basement Membrane; Biopsy; Cell Division; Humans; Immunoenzyme Techniques; Keratins; Psoriasis; Skin

Keratin-agarose plate and keratin-polyacrylamide enzymography methods were developed to demonstrate proteolytic digestion of epidermal keratin. By applying these methods, keratin hydrolase was purified from Tris-buffered saline extract of psoriatic scales by 50% ammonium sulfate precipitation, passage through a lysine-Sepharose column, DEAE-Sepharose, Sephacryl S-200, high-performance cation-exchange chromatography on Mono S, and aprotinin-Sepharose affinity chromatography. The final preparation demonstrated a single protein band at molecular weight 30,000 judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Furthermore, in keratin-polyacrylamide slab gels, the purified enzyme preparation showed a translucent band at molecular weight 30,000, indicating keratin digestion. Keratin hydrolase digested reassembled epidermal keratin as well, whereas it had no effect on guinea pig hair keratin. The enzyme demonstrated a high level of hydrolytic activity on Ile-Pro-Arg-p-nitroanilide and other peptidyl arginine substrates, while it showed a low level of activity on Val-Leu-Lys-p-nitroanilide, and no activity on Arg-Pro-Tyr-p-nitroanilide, Glu-Pro-Val-p-nitroanilide, or Ala-Ala-Ala-p-nitroanilide. The keratin hydrolase was a serine proteinase, inactivated by diisopropylfluorophosphate, phenylmethylsulfonyl fluoride, tosyl-lysyl-chloromethyl ketone, antipain, leupeptin, soybean trypsin inhibitor, aprotinin, and p-aminobenzamidine. The keratinolytic activity was not detected in normal epidermal extract.

Topics: Adult; Animals; Electrophoresis, Polyacrylamide Gel; Female; Guinea Pigs; Humans; Keratins; Male; Middle Aged; Peptide Hydrolases; Protease Inhibitors; Psoriasis; Sepharose; Skin

Products of the HLA-D gene, the la or DR antigens, have been shown to control interactions between certain cells with immune functions. Three HLA-D alleles have been reported to be associated with a markedly increased relative risk in psoriatic individuals. We report the existence of apparently unique anatomic interactions between lymphocytes and basal keratinocytes, between Langerhans' cells and basal keratinocytes, and between Langerhans' cells and lymphocytes, deduced on the basis of (1) characteristic cytoplasmic processes extending from one cell into the cytoplasm of adjacent cells, with frequent absence of the intervening plasma membrane at the apexes of these processes, and (2) intimate apposition of the plasma membranes between the interaction cells over a large surface area. These interactions were noted in five of ten untreated psoriatic patients (three of four patients with Koebner's phenomenon) and in one of six treated psoriatic patients, but in none of 17 controls. Intercellular space abnormalities, probably secondary to excessive proteolytic enzyme release, and basal keratinocyte herniations in psoriasis may result from these anatomic interactions.

Topics: Adult; Cell Communication; Cytoplasm; Female; Histocompatibility Antigens Class II; Humans; Intracellular Fluid; Keratins; Langerhans Cells; Lymphocytes; Male; Microscopy, Electron; Middle Aged; Psoriasis

Frozen sections of punch biopsies from normal epidermis and psoriatic involved and uninvolved epidermis have been examined immunocytochemically using a panel of anti-keratin monoclonal antibodies with various specificities in the skin. Since psoriasis is thought to involve hyperproliferative expansion of the basal compartment from one to about three cell layers in thickness, the samples were screened with antibodies to intermediate filament determinants associated with basal cells, suprabasal cells and hyperproliferating keratinocyte-derived cell lines, respectively. The basal-suprabasal division was observed to be intact, with only one layer of basal cells demarcated by the specific antibodies used under all circumstances. This suggests that (a) psoriatic "basal cell hyperproliferation' may not specifically involve the basal cell compartment containing the stem cells, but rather a population of amplifying transit cells which are predominantly suprabasal, and that (b) while keratinocyte differentiation begins as the cells lose contact with the basal lamina, the first stages at least of differentiation are not dependent on the loss of the capacity to divide.

Topics: Antibodies, Monoclonal; Antigens; Cell Differentiation; Cells, Cultured; Epidermis; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Keratins; Psoriasis

Topics: Cell Nucleus; Dose-Response Relationship, Radiation; Epidermis; Humans; Hyperplasia; Keratins; Lentigo; Melanocytes; Melanosis; Photochemotherapy; Psoriasis; PUVA Therapy; Skin Neoplasms

A skin equivalent model has been used to fabricate tissues with psoriatic and normal cells. Psoriatic fibroblasts can induce hyperproliferative activity in normal keratinocytes. The psoriatic epidermis from lesions continues to proliferate at high rates for at least 15 days in this model, and normal fibroblasts are unable to suppress this hyperproliferation. The primary defect in psoriatic skin may reside in the dermal fibroblast.

Topics: Adult; Animals; Collagen; Epidermal Cells; Female; Fibroblasts; Humans; In Vitro Techniques; Keratins; Male; Mice; Mice, Nude; Psoriasis; Skin

Topics: Calmodulin; Cell Division; Humans; Keratins; Psoriasis; Skin; Skin Diseases

psi-3 is a monoclonal antibody that recognizes a 135,000 molecular weight structural component of maturing keratinocytes in psoriasis (the psi-3 antigen) but fails to bind to any constituent of keratinocytes in normal epidermis. This paper describes the occurrence of the psi-3 antigen in a variety of dermatopathologic conditions using immunoperoxidase (biotin-avidin-peroxidase) and immunofluorescence methods which show excellent concordance. In 35 of 36 specimens of psoriasis vulgaris, psi-3 antibody consistently immunolabels the cytoplasm of keratinocytes above the basal layer. At the edges of psoriatic plaques, psi-3 antibody staining extends for a variable distance into lesion-free epidermis. A similar pattern has been found in a certain number of other conditions described in the paper, including squamous cell carcinoma and condyloma acuminatum, but not Darier's disease, basal cell carcinoma, nor lamellar ichthyosis. In all but one condition, the outermost or basal layer of cells is never stained. The only disease in which the lowermost cell layer is stained is a lichen planus-like lesion. The occurrence of psi-3 antigen cannot be correlated with any histologic feature of psoriasis such as acanthosis, loss of the granular layer, or hyperproliferation. The antigen appears to be a unique keratinocyte constituent which is expressed in certain pathologic conditions and which is not detected by any other histologic or immunophenotyping method. It is a potentially valuable addition to the panel of antibodies available for characterizing epithelial cells.

Topics: Antibodies, Monoclonal; Antigens; Biopsy; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Keratins; Psoriasis; Skin; Skin Diseases; Skin Neoplasms; Staining and Labeling

A murine hybridoma secreting an IgM monoclonal antibody (KL3) was produced by cell fusion of mouse myeloma cells with spleen cells from mice immunized with human epidermal keratins. On normal human epidermis KL3 stained the intercellular spaces from the stratum germinatum to the stratum granulosum with a fluorescence intensity increasing from the basal layer to the upper layers. Basal cells were not stained on the side facing the basement membrane. About 90% of free keratinocytes isolated after trypsinization were labelled by KL3 in a punctate staining. Immunoelectron microscopy allowed us to show that the antigen recognized by KL3 was exclusively localized on the keratinocyte membrane especially in the desmosomal plaques. KL3 reactivity was not modified by preincubation of skin sections with lectins showing a selective intercellular labelling of upper layers of epidermis or pemphigus antisera, nor by adsorption of the antibody on NP40 soluble proteins of the epidermis. Though KL3 reactivity was completely abolished after adsorption of purified keratins, no immunological reactivity of KL3 was detected with epidermal keratin polypeptides blotted on nitrocellulose paper. In psoriatic epidemis and epidermal tumors KL3 reactivity was drastically modified. These results suggest that KL3 recognized a keratinocyte membrane antigen implied in the epidermal differentiation process.

Topics: Animals; Antibodies, Monoclonal; Antigens, Surface; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Cell Differentiation; Cell Membrane; Epidermal Cells; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; Keratins; Macaca fascicularis; Mice; Psoriasis; Rabbits; Skin; Skin Diseases; Skin Neoplasms; Warts

Calcium has been shown to regulate the proliferation of epidermal keratinocytes in vitro. We became interested in the role of the calcium binding protein, calmodulin, in hyperproliferative, low calcium regulated keratinocytes in vitro and in the in vivo hyperproliferative state, psoriasis. Calmodulin levels were measured by radioimmune assay in neonatal mouse keratinocytes grown in 0.02 mM calcium (hyperproliferative) and 1.2 mM calcium (normal) media, and in cells that had been grown in low calcium medium and then switched to normal calcium. On a whole culture basis the normal cells had more calmodulin than the low calcium cells. However, when low calcium monolayers were compared to the normal basal monolayer, the low calcium hyperproliferative cells had more calmodulin. Cells that were switched from 0.02 mM calcium to 1.2 mM calcium showed increasing calmodulin levels over time. Psoriatic plaques contained 2-3 times more calmodulin than the skin of normal controls when examined on a per micrograms of DNA, per micrograms of protein, and per gram of wet weight basis. Adjacent uninvolved psoriatic skin also had significantly elevated calmodulin levels in all data bases except per microgram of protein/cm2. These data suggest that increased calmodulin levels are associated with epidermal hyperproliferation and/or with the state of differentiation.

Topics: Animals; Calcium; Calmodulin; Cell Division; Cells, Cultured; Epidermal Cells; Epidermis; Humans; Keratins; Mice; Mice, Inbred BALB C; Psoriasis

Clinically uninvolved psoriatic epidermis shows increased DNA synthesis in vivo. We have studied the DNA synthesis of cultured keratinocytes from uninvolved psoriatic skin. Trypsinized epidermal cells were plated on plastic dishes pre-coated with bovine collagen type I. In initial studies, normal human serum was found to be superior to fetal bovine in supporting the growth of human epidermal keratinocytes. Furthermore, keratinocyte cultures established in the presence of normal human serum produced large keratin proteins (68,000 daltons) indicating that the terminal steps in cell differentiation can occur in vitro. In subsequent experiments keratinocyte cultures were grown in medium supplemented with 10% normal human serum. Confluent cultures of keratinocytes from uninvolved psoriatic epidermis had an increased DNA synthesis determined both as the incorporation of [3H]thymidine and as the autoradiographic labelling index. The DNA synthesis of both normal and psoriatic keratinocyte cultures increased in response to incubation in medium with 10% psoriatic serum. The ability of keratinocytes from uninvolved psoriatic epidermis to maintain an increased DNA synthesis suggests the presence of an inherent defect within the population of epidermal keratinocytes in psoriasis. Such a culture system can be used as an in vitro model for the study of psoriasis.

Topics: Adult; Blood; Cells, Cultured; Culture Media; DNA; Epidermis; Humans; Keratins; Molecular Weight; Psoriasis; Thymidine

Extensive morphological and biochemical studies were carried out on two sibling cases of ichthyosis linearis circumflexa. The condition was found to be similar to psoriasis in the following ways: effectiveness of PUVA therapy, psoriatic changes on light and electron microscopy, high urinary polyamine levels, elevation of several enzyme activities in the scales, and a remarkable change in keratin molecules. These results may reflect an increased epidermal proliferation with a reduced epidermal transit time, as occurs in psoriasis.

Topics: Adolescent; Adult; Amino Acids; Electrophoresis, Polyacrylamide Gel; Female; Humans; Ichthyosis; Keratins; Male; Microscopy, Electron; Polyamines; Psoriasis; Skin

Isoquinoline has been identified as a component of coal tar which causes interfollicular regions of parakeratotic stratum corneum in mouse tail epidermis to become orthokeratotic, with concomitant production of a granular layer. In this respect it behaves similarly to coal tar itself, and isoquinoline may contribute to the anti-psoriatic activity of coal tar.

Topics: Animals; Coal Tar; Cricetinae; Epidermis; Isoquinolines; Keratins; Mice; Mice, Inbred Strains; Psoriasis

A histological comparison was made between normal mouse tails and those treated with crude coal tar, and the effect of crude coal tar on the keratin profile of the living cells of treated animals was examined. The prophylactic effect of crude coal tar on the neonatal mouse tail is described. The variation in the anatomical site of prekeratin of the dorsal and tail epidermis of the mouse is reported. These results are discussed with reference to the use of the mouse tail as a model for screening drugs for the treatment of psoriasis.

Topics: Animals; Coal Tar; Disease Models, Animal; Keratins; Male; Mice; Mice, Inbred Strains; Molecular Weight; Protein Precursors; Psoriasis; Skin

Human keratinocytes were investigated for the presence of single cilia. Almost all basal keratinocytes were found to carry a single cilium in normal, occluded, and psoriatic skin. The ciliary structure was progressively reduced in keratinocytes approaching the surface. No remnants of the ciliary apparatus were found in the granular layer. In one case of nickel-allergic dermatitis (patch test), the keratinocytes had lost their cilia; the significance of this surprising finding remains to be elucidated.

Topics: Adult; Cilia; Dermatitis, Contact; Epidermal Cells; Epidermis; Humans; Keratins; Microscopy, Electron; Nickel; Psoriasis

Composite and intranuclear keratohyalin granules were observed in an electron-microscopic study of geographic tongue of psoriasis. The significance of intranuclear keratohyalin is not known, but it seems to be related to the disturbed proliferation and differentiation of keratinocytes in psoriasis.

Topics: Cell Nucleus; Cytoplasmic Granules; Glossitis, Benign Migratory; Humans; Keratins; Microscopy, Electron; Mouth Mucosa; Psoriasis; Tongue

The keratin polypeptide composition of uninvolved psoriatic epidermis is the same as normal epidermis. In contrast, involved epidermis from psoriasis contains significant amounts of two keratin polypeptides, molecular weight (MW) 56 and 50 kilodaltons (KD). Involved epidermis contains decreased amounts of keratins MW 69, 66.5 and 65.2 KD. Slices of involved psoriatic epidermis incorporate [35S]methionine into all the keratins including those of MW 56 and 50 KD. Unexpectedly, we found that [35S]methionine is incorporated into 56 and 50 KD keratins in uninvolved epidermis even though these keratins are not abundant enough to be seen in Coomassie stained gels. The two-dimensional patterns of iodinated tryptic peptide digests of keratin MW 50 KD from uninvolved, involved and normal epidermis are identical. We conclude that keratins of MW 56 KD and 50 KD are abundant in involved psoriatic epidermis. They are not present in normal or uninvolved psoriatic epidermis, but are rapidly synthesized when normal epidermis (neonatal foreskin) or uninvolved psoriatic epidermis are removed and placed in organ culture. The two keratins may be markers of increased cell proliferation or injury.

Topics: Aged; Autoradiography; Cells, Cultured; Electrophoresis, Polyacrylamide Gel; Humans; Infant, Newborn; Keratins; Male; Methionine; Molecular Weight; Peptide Fragments; Psoriasis; Skin; Sulfur Radioisotopes

We studied the localization of p230, an immunoanalogue of erythroid alpha-spectrin, in normal and psoriatic human epidermis and in cultured human keratinocytes. In immunofluorescence microscopy of frozen sections of normal skin a bright cytoplasmic staining was seen in the cortical area of the keratinocytes. Similar staining was also seen in lesional and uninvolved areas of psoriatic epidermis. The pericytoplasmic localization of p230 could also be seen in cultured human keratinocytes: a lamina-like reticular staining was seen mostly confined to the ventral cytoplasmic aspect and to junctional areas of the cells. Immunoblotting of electrophoretically separated polypeptides of epidermal cells revealed a distinct polypeptide of Mr 230 kD. The results indicate that alpha-spectrin-like polypeptides form a major cytoskeletal framework in human epidermal cells in both normal and psoriatic skin.

Topics: Cells, Cultured; Epidermal Cells; Epidermis; Fluorescent Antibody Technique; Humans; Keratins; Peptides; Psoriasis

To establish a method for separating different keratinocyte subpopulations in the epidermis, we studied the specificity of monoclonal antibody 4F2 for keratinocytes. Preliminary screening experiments had previously demonstrated 4F2 reactivity with the epidermis. 4F2 reacted with a subpopulation (19.29 +/- 5.23%) of human epidermal cells in suspension. The membrane antigen identified by 4F2 continues to be expressed by cultured keratinocytes. In frozen tissue section using an indirect immunofluorescence technique, the 4F2-positive cells in the basal layer are sharply demarcated from the negative suprabasilar layers. Even in the hyperproliferative state of psoriasis, the 4F2 reactivity is confined to the basal layer. Cell suspensions of psoriatic epidermis demonstrated a greater percentage of reactivity with 4F2 (49.51% +/- 6.50%), probably reflecting the expanded population of basal layer cells. Monoclonal 4F2, therefore, reacts with a membrane antigen present on basal keratinocytes, and provides a probe for use in the isolation of the basal keratinocyte subpopulation. Thus, this antibody should be useful in studies of normal and aberrant differentiation of the epidermis.

Topics: Antibodies, Monoclonal; Antigens, Surface; Cell Division; Epidermis; Fluorescent Antibody Technique; Humans; Keratins; Psoriasis

Topics: Cytoplasm; Epidermis; Humans; Keratins; Psoriasis

Topics: Electrophoresis; Gingiva; Humans; Keratins; Psoriasis; Skin

We have isolated a murine monoclonal antibody, called psi-3, which immunolabels maturing keratinocytes in psoriatic skin but not in normal epidermis. The staining is cytoplasmic and is not extractable with 1% Triton X-100, which suggests that the psi-3 antigen is a structural component of the keratinocyte. Neither basal cells nor invading inflammatory cells are stained in psoriatic skin and the antigen appears to be associated specifically with maturing and not proliferating keratinocytes. Keratinocytes cultured in vitro from skin from nonpsoriatic individuals display the antigen in a granular pattern in differentiated cells. The antigen is also expressed after tape-stripping of normal skin and, therefore, represents an inducible product of normal keratinocytes. The antigen is destroyed by proteinase K and appears to be a protein. On discontinuous sodium dodecyl sulfate-gel electrophoresis, the antigen has been found to have a molecular weight of 135,000. The psi-3 antigen is interpreted as a new keratinocyte product expressed in psoriasis, culture, wound healing, and certain other pathologic skin conditions. The synthesis of such a new antigen would not be expected if keratinocyte maturation in psoriasis is a truncated version of the normal system and supports the hypothesis that psoriatic keratinocytes are following an alternative pathway. Results using experimental injury suggest that the psoriatic pathway is normally expressed during wound healing.

Topics: Animals; Antibodies, Monoclonal; Antigens; Cells, Cultured; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Hybridomas; Keratins; Mice; Mice, Inbred BALB C; Psoriasis; Skin; Wound Healing

We describe the effect of two polyamine antimetabolites on polyamine and macromolecule synthesis of cultured human keratinocytes obtained by suction blisters from normal skin and the uninvolved skin of psoriatic patients. The concentrations of spermidine and spermine steadily increased during the culture of normal keratinocytes in vitro, whereas the putrescine concentration showed only a transient rise at the beginning of the active growth phase. Treatment with difluoromethylornithine decreased the concentrations of putrescine and spermidine in both normal and uninvolved psoriatic keratinocytes, but had no effect on either DNA or protein synthesis. Methylglyoxal bis(guanylhydrazone) marginally decreased the levels of spermidine and spermine and significantly inhibited the DNA and protein synthetic activities. Pretreatment of uninvolved psoriatic keratinocytes with difluoromethylornithine enhanced the accumulation of methylglyoxal bis(guanylhydrazone), resulting in a profound inhibition of cellular macromolecule synthesis. This synergistic effect was not seen in normal keratinocytes. Thus, although no statistically significant difference was observed between the cells derived from normal and uninvolved psoriatic epidermis, the psoriatic keratinocytes appeared to be more sensitive to the action of polyamine antimetabolites. The inhibition of DNA and protein synthesis by methylglyoxal bis(guanylhydrazone) was prevented by concomitant treatment with spermidine.

Topics: Antimetabolites; Cell Division; Cells, Cultured; DNA; Eflornithine; Guanidines; Humans; Keratins; Mitoguazone; Ornithine; Protein Biosynthesis; Psoriasis; Putrescine; Skin; Spermidine; Spermine

Topics: Epidermal Cells; Humans; In Vitro Techniques; Interferon Type I; Interferon-gamma; Keratins; Psoriasis; T-Lymphocytes

Quantification and characterization of [3H]-fucose-labeled cell surface glycoproteins are reported. Two approaches have been compared; first the analysis of glycoprotein shed spontaneously into the medium during incubation of keratinocytes in vitro, and second the study of material released by exposure of the cell to proteolytic enzymes. It is shown that psoriatic keratinocytes "shed" glycoprotein more rapidly than normal, although the material is of similar molecular weight (mainly "biantennary" transferrin type glycopeptides). By contrast, the percentage of glycoprotein released by proteolysis of psoriatic keratinocytes is normal, but the molecular weight distribution of the labeled glycopeptides is markedly altered. The abnormal turnover and composition of fucose-labeled glycoproteins from the cell surface may be related to the loss of growth control in psoriatic epidermis.

Topics: Cell Membrane; Glycopeptides; Glycoproteins; Humans; Keratins; Membrane Proteins; Peptide Fragments; Psoriasis; Skin; Trypsin

High-resolution electrophoresis has been used to extend previous observations on the polypeptide composition of keratins in psoriatic epidermis. We have compared psoriatic scale keratins with normal and with scale extracts from several different epidermal disorders. Uninvolved psoriatic epidermis contained prekeratin and keratin of normal profile (68, 60, 58, 52 kDa and 66, 58, 55 kDa, respectively). Prekeratin from involved psoriatic epidermis showed a variable quantitative reduction in the 68-kDa polypeptide and an altered expression of smaller polypeptides (Mr 40 000-55 000). Keratin from the psoriatic lesion was abnormal and appeared 'prekeratin-like'. Keratin from the involved stratum corneum of patients with seborrhoeic eczema. Darier's disease and common dandruff were also similar to prekeratin, but that from ichthyosis and toxic epidermal necrolysis was normal. These results suggest that psoriatic keratinocytes have a defective but variable expression of prekeratin polypeptides. Furthermore, the differentiation-linked modification of prekeratin to keratin is defective in psoriasis, a phenomenon found in other hyperkeratotic epidermal disorders.

Topics: Adolescent; Adult; Aged; Child; Electrophoresis, Polyacrylamide Gel; Female; Humans; Keratins; Male; Middle Aged; Molecular Weight; Protein Precursors; Psoriasis; Reference Values; Skin

Topics: Adult; Aged; Cell Membrane; Desmosomes; Epidermis; Etretinate; Freeze Fracturing; Humans; Keratins; Middle Aged; Psoriasis; Tretinoin

Keratin was extracted from normal human horny cells of the leg, calluses of the sole, and psoriatic scales. After dissociation in sodium dodecyl sulfate the polypeptides were separated by Laemmli's gel electrophoresis method and their molecular weights and relative amounts determined. Normal horny cells contained 3 polypeptide chains of Mr 67K, 59K, and 57K, while those of callus contained 9 polypeptides of Mr 67K, 66K, 63K, 62K, 58K, 54K, 52K, 48K, and 45K. In both cases all keratin polypeptides participated in filament reassembly in vitro and were recovered from the filaments. In psoriatic scale keratin, 7 prominent polypeptides were detected having Mr 67K, 59K, 57K, 50K, 48K, 42K, and 40K. The 67K polypeptide could not be recovered from reassembled filaments. Ultrastructural studies revealed that these filaments are imperfect and readily aggregate into thick fibrils. These observations indicate that there are significant differences in composition of keratin of normal horny cells, calluses, and psoriatic scales.

Topics: Callosities; Electrophoresis, Polyacrylamide Gel; Epithelium; Humans; Keratins; Peptides; Psoriasis

The variations in the expression of epidermal keratins occurring during retinoid therapy were studied in patients with psoriasis before and during oral administration of etretinate (aromatic retinoid Ro 10-9359) and compared with normal epidermis. Sodium dodecyl sulfate polyacrylamide gel electrophoresis densitometric readings were performed on keratins extracted from epidermal cells obtained through trypsinization of skin specimens. Epidermal cells were also tested by immunofluorescence (IF) for the presence of BMZ antigens as markers of basal cells and for the presence of TK and KP (67 K, 63 K, and 55 K) with sera with antibodies against BMZ antigens and specific antisera for TK and KP. In psoriasis-involved epidermis, SDS-PAGE anaLysis showed lower amounts of 67 K and increased amounts of 63 K and 55 K, as compared with normal epidermis. Low proportions of cells expressing by IF the 67 K and the 63 K were also noted, with a defective expression of these two KP by suprabasal keratinocytes in psoriasis-involved epidermis. During etretinate administration, a return toward the normal electrophoretic pattern and a correction of the defective cellular expression of these two KP were obtained parallel with clinical improvement. These findings indicate the presence in involved psoriatic epidermis of a population of suprabasal keratinocytes that do not express the high-molecular-weight KP and show a normalization of the relative proportions of the KP with etretinate.

Topics: Adolescent; Adult; Antigens; Electrophoresis, Polyacrylamide Gel; Epidermis; Etretinate; Fluorescent Antibody Technique; Humans; Keratins; Male; Middle Aged; Peptides; Psoriasis; Tretinoin

Topics: Adult; Aged; Child; Electrophoresis, Polyacrylamide Gel; Epidermis; Female; Humans; Keratins; Male; Middle Aged; Peptides; Psoriasis; Sodium Dodecyl Sulfate

Surface glycoproteins of cultured human keratinocytes from normal skin and uninvolved psoriatic epidermis, isolated by the suction blister method, were studied by two different methods. Cells were cultured on collagen-coated culture dishes and showed a fibrillar keratin-specific staining by immunofluorescence. Surface labelling experiments using the neuraminidase/galactose oxidase/sodium borohydride method (which labels the penultimate galactose moieties of glycoproteins) revealed one major glycoprotein with Mr 53 kD (kilodaltons) both in normal keratinocytes and in keratinocytes from uninvolved psoriatic skin. The periodate/sodium borohydride method (which labels the terminal sialic acids in glycoproteins) by contrast revealed three major glycoproteins, with Mr 53 kD to 63 kD, in normal keratinocytes but only a single major glycoprotein, with Mr 53 kD, in keratinocytes from uninvolved psoriatic skin. Treatment of cultured keratinocytes with etretinate appeared to restore the normal pattern of surface glycoproteins in uninvolved psoriatic keratinocytes.

Topics: Cells, Cultured; Epidermis; Etretinate; Glycoproteins; Humans; Keratins; Membrane Proteins; Psoriasis

Topics: Administration, Oral; Electrophoresis, Polyacrylamide Gel; Epidermis; Etretinate; Humans; Keratins; Molecular Weight; Psoriasis

The glycocalyx of epidermal keratinocytes from psoriatic patients has been investigated by means of lectins. Striking changes were found in the levels of glucose and/or mannose (concanavalin A) and of N-acetylglucosamine and/or sialic acid (wheat germ agglutinin) on the surface of cells from the psoriatic lesion. Smaller but significant changes were seen in the clinically uninvolved epidermis of the patient. A marked increase in the affinity of the cell surface for Ulex europus agglutinin (fucose-specific) confirms our previous reports of structural alterations in fucose-containing oligosaccharides in psoriasis.

Topics: Binding Sites; Carbohydrate Metabolism; Cell Membrane; Humans; Keratins; Lectins; Psoriasis; Skin

Polyacrylamide gel electrophoretic investigations were carried out on solubilized proteins from psoriatic and normal stratum corneum obtained by adhesive tape stripping. The proteins in the scales adhering to the tape were solubilized by incubating the tape in 1% sodium dodecyl sulphate (SDS) solution. The electrophoretic behaviour of these solubilized proteins on SDS-polyacrylamide gel was compared with the alpha-fibrous proteins (keratin) of callus. The proteins isolated from callus of normal human heel showed six main bands which were similar to those of the keratin isolated by the 8 M urea-mercaptoethanol method. The lesional skin of forty-five psoriatic patients consistently showed nine main bands on polyacrylamide gels, but only two main bands were observed in the non-lesional, non-heel skin. Six of these nine bands had mobilities and relative intensities almost identical with those of alpha-keratin extracted by the mercaptoethanol method, but the other three bands had greater mobilities on the gels. These results suggest that this technique may have considerable potential for studying changes in alpha-keratin in patients with psoriasis and other disorders of keratinization.

Topics: Callosities; Electrophoresis, Polyacrylamide Gel; Humans; Keratins; Methods; Proteins; Psoriasis; Skin

Intubation of unfixed and unfrozen slices of skin in diaminobenzidine allows visualization of a peroxidatic activity in perinuclear envelope of suprabasal keratinocytes undergoing orthokeratotic differentiation. Basal keratinocytes and melanocytes are always negative. This enzyme is absent in mucous and parakeratotic (psoriatic) differentiation. Mitochondria are also strongly stained by this technique and it was shown that the number of epidermal mitochondria is greatly increased in psoriatic lesions.

Topics: 3,3'-Diaminobenzidine; Benzidines; Cytochrome-c Peroxidase; Endoplasmic Reticulum; Histocytochemistry; Humans; Keratins; Mitochondria; Nuclear Envelope; Peroxidases; Psoriasis; Skin

Topics: Adhesives; Humans; Keratins; Proteins; Psoriasis; Skin; Sulfhydryl Compounds

The effect of cellular pathology and keratinization of skin and nasal cells upon binding of Staphylococcus aureus were examined. Adherence with epithelial cells obtained from either the skin or nasal mucosa of patients with atopic dermatitis was greater than that observed with normal cells (p less than 0.001); the difference in adherence between psoriatic and normal cells was not statistically significant. Tested nasal cells were microscopically differentiated into 4 general types based on stage or layer of keratinization: spinous, low granular, high granular, and keratin. The degree of adherence was related to the progress of keratinization. Data indicated the existence of 2 types of receptors for S. aureus on nasal cells: One, present upon both granular and fully keratinized cells, is not blocked by teichoic acid and appears responsible for the higher bacterial counts on atopic cells; the second is found on keratinized cells only and is susceptible to teichoic acid.

Topics: Adhesiveness; Bacterial Physiological Phenomena; Dermatitis, Atopic; Epithelium; Humans; Keratins; Nasal Mucosa; Psoriasis; Skin; Skin Physiological Phenomena; Staphylococcus aureus

Topics: Acne Vulgaris; Humans; Isotretinoin; Keratins; Psoriasis; Skin Diseases; Skin Neoplasms; Tretinoin; Vitamin A

Topics: Cytoskeleton; Humans; Keratins; Molecular Weight; Protein Precursors; Psoriasis; Skin; Skin Diseases

We report the uptake of four labelled sugars by keratinocytes isolated from normal epidermis, psoriatic 'uninvolved' skin and psoriatic lesions. Our findings include the following: (1) The rate of uptake of all sugars by the psoriatic lesion is increased. (2) This abnormally high uptake diminishes dramatically during 22 h incubation in vitro. (3) There is a striking abnormality in the metabolism of fucose by psoriatic keratinocytes; our data suggest an increased rate of incorporation of fucose into glycoconjugates.

Topics: Carbohydrate Metabolism; Cell Membrane; Epidermis; Fucose; Glycolipids; Glycoproteins; Humans; In Vitro Techniques; Keratins; Proteoglycans; Psoriasis; Skin; Time Factors

Topics: Anthracenes; Anthralin; Cell Division; Epidermis; Humans; Keratins; Psoriasis

A substantial proportion (20-50%) of radioactive sugar incorporated into glycoconjugates by normal human keratinocytes was soluble in chloroform-methanol; using (14C)-galactose as precursor about half of this fraction was neutral lipid. The incorporation of labelled sugars into the lipid fraction was consistently increased in keratinocytes derived from psoriatic lesions. The abnormality in fucose metabolism which we reported previously has been confirmed. In particular we have shown that the molecular weight of fucosylated glycopeptides appears to be abnormally high in the untreated psoriatic lesion, possibly reflecting an increased degree of branching. In psoriatic uninvolved epidermis and in treated psoriatic lesions the situation is reversed, the molecular weight being significantly lower than normal.

Topics: Cell Membrane; Chromatography, Gel; Chromatography, Thin Layer; Glycolipids; Glycopeptides; Glycoproteins; Glycosaminoglycans; Humans; Keratins; Membrane Lipids; Membrane Proteins; Proteoglycans; Psoriasis; Skin

Keratin proteins were extracted from scales of normal skin, clinically active psoriatic lesions, and atopic dermatitis. Filaments prepared by in vitro assembly upon dialysis of the proteins against a low ionic strength buffer were comparatively characterized by electron microscopy, SDS gel electrophoresis, and amino acid analysis. Filaments formed using keratin obtained from the skin of normal individuals were thin and wavy, whereas those formed using keratin isolated from the scales of psoriatic patients were straight and showed a tendency to assemble side by side. Filaments of atopic dermatitis were indistinguishable from those of normal individuals. Filaments of both normal and atopic dermatitis contained the protein band of 67,000 daltons, which was absent in filaments of psoriasis. In contrast, 2 protein bands of 54,000 and 57,000 daltons were only detectable in psoriasis. Amino acid analysis of these filaments further demonstrated that filaments of psoriasis differ from those of normal individuals in that they have a glycine content that is 60% of normal.

Topics: Amino Acids; Dermatitis, Atopic; Humans; Keratins; Molecular Weight; Proteins; Psoriasis; Skin

When human and rat epidermis are exposed to 32P-orthophosphoric acid, labeled phosphate is incorporated into several proteins. The pattern of phosphorylation is identical whether the isotope is delivered in vivo or in vitro. The predominant phosphorylated proteins are insoluble in Tris-HCl buffer but soluble in SDS-beta-mercaptoethanol. They migrate in SDS-polyacrylamide gels with apparent molecular weights between 45,000 and 65,000. When analyzed by two-dimensional gel electrophoresis, the labeled phosphoproteins co-migrate with keratins isolated from human callus. Serine is the phosphate acceptor in these proteins. The pattern of phosphorylation of these SDS-beta-mercaptoethanol soluble epidermal proteins is not changed in basal cell carcinoma, icthyosis vulgaris, Kyrle's disease or Netherton's syndrome. The pattern is altered in psoriasis. Others have demonstrated that a 63,000 molecular weight protein is absent from active psoriatic lesions. We have found that a 63,000 molecular weight phosphoprotein is present in uninvolved skin but absent in the psoriatic plaques.

Topics: Animals; Bony Callus; Electrophoresis, Agar Gel; Epidermis; Humans; In Vitro Techniques; Keratins; Molecular Weight; Phosphorylation; Psoriasis; Rats; Skin Diseases

The sodium lauryl (dodecyl) sulfate polyacrylamide gel electrophoresis (SLS PAGE) patterns of keratin polypeptides were followed in 29 patients with psoriasis who were treated with middle wavelength ultraviolet (UV) radiation (UV B), oral methoxsalen, and long wavelength UV radiation (PUVA), or systemic antipsoriatic therapy. The keratin patterns from the Malpighian layer reverted to normal in several weeks, while those of the stratum corneum keratin reverted more slowly, often after clinical clearance of the lesions. These data may indicate that therapy should not be discontinued until there is production of normal stratum corneum polypeptides.

Topics: Electrophoresis, Polyacrylamide Gel; Epidermis; Furocoumarins; Humans; Keratins; Methoxsalen; Peptides; Photochemotherapy; Psoriasis; Sodium Dodecyl Sulfate

Electron microscope autoradiography has been used to compare the fucose-containing glycoproteins of normal and psoriatic epidermis. Normal keratinocytes and uninvolved psoriatic cells have most of their fucose on the plasma membrane, whereas involved psoriatic cells retain more within the cytoplasm. The fucose distribution in other epidermal cell types is also described.

Topics: Autoradiography; Cell Membrane; Cytoplasm; Fucose; Glycoproteins; Humans; Keratins; Microscopy, Electron; Organ Culture Techniques; Psoriasis; Skin

Topics: Humans; Keratins; Langerhans Cells; Monocytes; Psoriasis; Research Design

Topics: Adult; Child; Epidermis; Humans; Keratins; Microscopy, Electron; Psoriasis

Topics: Cells, Cultured; Culture Techniques; Electrophoresis, Polyacrylamide Gel; Humans; Keratins; Psoriasis; Skin

The structure of single filaments of horny cells of normal and psoriatic epidermis was studied in the electron microscopy in situ and in filaments reconstituted from urea extracts by dialysis, against low ionic strength buffer. It was found that both in situ and reconstituted filaments consist of 20 A protofibrils. In filaments of normal horny cells the protofibrils seem to form a rope-like structure while in those of psoriatic horny cells protofibrils appear randomly arranged. The filaments reconstituted from extracts of psoriatic scales range in width from 90 to 500 A, indicating a defect in lateral assembly of protofibrils. Numerous small particles are detectable at high magnification in extracts of both normal and psoriatic horny cells, which are about 20 A wide and 200 A long. These particles are consisted to be the basic structural units from which the protofibrils are formed, by end-to-end junctions.

Topics: Callosities; Cytoskeleton; Epidermis; Humans; Keratins; Microscopy, Electron; Psoriasis

The chemistry and structure of the epidermal alpha-keratin extracted from the skin of patients with a variety of disorders of keratinization have been investigated using biochemical, biophysical, and electron microscopic techniques developed for the characterization of normal mammalian epidermal keratin. Generally, the alpha-keratin polypeptides of the diseased epidermis differed from those of uninvolved epidermis or of normal volunteers in having varying numbers of polypeptide components of lower molecular weights, numerous free amino acids, higher contents of alpha-helix, and only limited facility for polymerization in vitro into native-type epidermal keratin filaments. As the alpha-helix-enriched fragments, which represent up to two-thirds of the polypeptide chains, isolated after limited tryptic digestion of the keratin filaments of normal, uninvolved, and involved epidermis, were physicochemically identical, it seems that the end-terminal non-alpha-helical regions of the polypeptides of diseased epidermis are abnormal. These differences may be a result of degradation or of altered protein synthesis.

Topics: Amino Acid Sequence; Callosities; Cytoskeleton; Epidermis; Humans; Keratins; Microscopy, Electron; Molecular Weight; Protein Conformation; Psoriasis; Skin Diseases

Topics: Amyloid; Amyloidosis; Carcinoma, Basal Cell; Epidermis; Fibroblasts; Histiocytes; Humans; Keratins; Psoriasis; Skin

It is our belief that any abnormality in keratinization is secondary to an increased rate of epidermal cell production and decreased transit time and research into the underlying nature of psoriasis should concentrate on the issue of whether the stimulus to increased epidermopoietic activity is dermally or epidermally derived.

Topics: Cell Division; Cell Membrane; Cytoplasm; Epidermis; Humans; Keratins; Microscopy, Electron, Scanning; Microvilli; Psoriasis; Skin

Topics: Cell Cycle; Cell Differentiation; Cell Division; Cell Membrane; Cyclic AMP; Epidermis; Humans; Keratins; Psoriasis

Cyclic AMP levels have been determined for the first time in isolated keratinocytes. Values were more reproducible than those reported using epidermal slices. Evidence is presented to show that damage to hormone receptors is minimal. Other observations include the following: (1) Keratinocytes from psoriatic lesions showed reduced 'resting' levels of cyclic AMP as well as a diminished response to adrenaline. (2) Cyclic AMP levels were maximal in the basal cells, falling dramatically in fully differentiated keratinocytes. (3) The topical application of a corticosteroid (fluocinolone acetonide) did not modulate the response of adenyl cyclase to hormonal stimulation.

Topics: Adolescent; Adult; Animals; Cattle; Cell Separation; Cyclic AMP; Epinephrine; Female; Fluocinolone Acetonide; Histamine; Humans; Keratins; Middle Aged; Psoriasis; Skin; Stimulation, Chemical; Time Factors

In a previous study, impedance measurements and phoreographic response were shown to quantify significant differences between involved and non-involved skin in psoriasis. The same techniques were used to objectivate the evolution on skin condition on patients treated with dioxyanthranol, difluprednate, and photochemotherapy associated with 8-methoxypsoralen. Each of these treatments was applied to four subjects. On patients treated with dioxyanthranol or difluprednate, impedance and phoreographic response return to "normal" values within 5--10 days and 1--2 days, respectively. It takes a longer time with photochemotherapy, and it is noteworthy that the treatment also alters the phoreographic response of non-involved skin. The changes observed in electrophysiological parameters are discussed in relation with recent ideas on pharmacology of antipsoriatic treatments and on dynamic properties of biological membranes. The results are in good correlation with clinical data and exemplify the usefulness of these methods in following the treatment kinetics.

Topics: Anthralin; Electric Conductivity; Fluprednisolone; Humans; Keratins; Kinetics; Methoxsalen; Mitotic Index; Photochemotherapy; Psoriasis; Skin; Skin Physiological Phenomena

A method is described for the preparation of isolated keratinocytes suitable for subsequent biochemical studies. Scanning electron microscopy showed that the maturation process is accompanied by an increase in cell size and a shortening and eventual loss of microvilli. Psoriatic keratinocytes are distinguishable by exhibiting longer microvilli at all levels of maturation.

Topics: Adolescent; Adult; Cell Membrane; Cell Separation; Humans; Keratins; Microscopy, Electron, Scanning; Microvilli; Middle Aged; Psoriasis; Skin

The polyacrylamide SDS electrophoretic pattern of protein extracted from the stratum corneum obtained by scraping the surface of involved skin of patients with psoriasis was different from that of uninvolved skin and normal controls. The pattern from superficial scales was similar to that of whole stratum corneum in the case of involved psoriatic epidermis but different in uninvolved and normal epidermis. These data indicate that the changes which are observed in the structural proteins during normal keratinization are not seen in involved psoriatic epidermis. In addition, the relative proportion of keratin polypeptides was different in involved psoriatic epidermis compared to normal skin. That these changes are not specific for psoriasis was shown by finding similar electrophoretic patterns with stratum corneum proteins from patients with other keratinizing disorders.

Topics: Electrophoresis, Polyacrylamide Gel; Humans; Keratins; Molecular Weight; Peptides; Psoriasis; Skin

Studies on the polypeptide chain compositions of human stratum corneum and callus have shown that characteristic modifications of the epidermal fibrous protein occur during normal eqidermal keratinization. The polypeptide chain composition of psoriatic scale differs from that of both normal stratum corneum and callus. A major constituent, the alpha-chain (Mr = 70 000) of the epidermal fibrous protein is either absent or greatly decreased in amount in psoriatic sacle. This chain is, however, present in normal amounts in the uninvolved epidermis of psoriatics and, during effective treatment, returns to normal levels in the previously involved areas. It is concluded that this is a major, reversible difference between involved psoriatic epidermis and uninvolved psoriatic or normal epidermis and that it most probably results from a defect in the synthesis of the alpha-chain.

Topics: Adolescent; Adult; Aged; Electrophoresis, Polyacrylamide Gel; Female; Humans; Keratins; Male; Middle Aged; Molecular Weight; Proteins; Psoriasis; Skin

Topics: Acid Phosphatase; Animals; Cathepsins; Chediak-Higashi Syndrome; Cyclic AMP; Dermatitis; Fabry Disease; Humans; Keratins; Lupus Erythematosus, Systemic; Lysosomes; Psoriasis; Skin; Ultraviolet Rays; Vacuoles; Vitamin A Deficiency

Light microscopy was used to examine the clinically normal palmar skin of 18 psoriatic patients (10 inactive and 8 active) and 18 non-psoriatic control patients. Histological changes in both epidermis and dermis vary with activity of the disease. The findings support the view that defective keratin formation precedes excessive proliferation of epidermal cells.

Topics: Biopsy; Hand Dermatoses; Humans; Keratins; Psoriasis; Skin

The rate and pattern of epidermal lipogenesis from [14C] glucose were measured in fifteen patients with psoriasis and three with lichen simplex, compared with twenty controls. In 'uninvolved' epidermis from psoriatic subjects the mean lipogenic rate was slightly raised, although the increase was not statistically significant. There was a positive correlation between overall lipogenic rate and the percentage of isotope appearing in free sterol, while the relative proportions of the other lipid classes were unchanged. By contrast, in control epidermis sterol percentage was negatively correlated with lipogenic rate. In psoriatic lesions total epidermal lipogenesis (per unit surface area) was raised compared with matched control 'uninvolved' epidermis. Also raised were percentage labelling of free sterol and of combined (free sterol and monoesters), and the free sterol: monoester ratio was increased. Similar findings were obtained with lesions of lichen simplex, suggesting that disturbed sterol metabolism may be a common feature in conditions of abnormal keratinization.

Topics: Adolescent; Adult; Aged; Esters; Female; Glucose; Humans; Keratins; Lipids; Male; Middle Aged; Neurodermatitis; Psoriasis; Skin; Sterols; Syndrome

Keratinosomes in psoriatic skin can be slightly larger than those in normal skin. Some seem to develop an unusual internal morphology, and some reach their full development while they are still within the immediate Golgi area. Observational evidence indicates that there may be more kerantinosomes in psoriatic skin than are generally found in normal skin. Keratinosomes appear extracellularly from the spinous layer through the horny layer. Some keratinosomes do not move to the outside of the cell and can be seen within cells of the horny layer. There appears to be an increased number of keratinosomes in and between the outer cells of sweat ducts. Keratinosomes are also present within the dark cells of the duct and the extracellular space between the dark cells contains material from keratinosomes.

Topics: Cytoplasmic Granules; Humans; Keratins; Psoriasis; Skin

Peripilar keratin casts are reported in three boys. All previous reports of peripilar keratin casts in children have apparently been in girls.

Topics: Adolescent; Child; Child, Preschool; Dermatitis, Seborrheic; Hair; Humans; Keratins; Male; Psoriasis; Skin Diseases

Electronmicroscopical investigation on 12 cutaneous biopsies of different localisation showed that the seborrhoic dermatitis is not comparable to a psoriatic tissue-reaction. Both, the cyto-morphological feature and the demonstrated localisation of the acid phosphatase are distinctly different from that of the psoriatic epidermis. The described changes are clearly more similar to the well known ultrastructural pictures of eczemateous reactions. In spite of the "eczema-like" ultrastructural picture seborrhoic dermatitis apparently can be separated from the allergic and irritant contact-dermatitis. Nevertheless, the epidermal alterations are also similar to chronic nummular eczema. They therefore are unspecific and do not allow any conclusion regarding the etiopathogenesis or nosological classification of this skin disease.

Topics: Acid Phosphatase; Basement Membrane; Cell Nucleus; Dermatitis, Contact; Dermatitis, Seborrheic; Desmosomes; Eczema; Endoplasmic Reticulum; Histocytochemistry; Humans; Keratins; Organoids; Phagocytes; Psoriasis; Skin; Vacuoles

The activity of ICDH(NAD) was measured in subcorneal and basal epidermal layers in 8 patients with psoriasis and in 7 healthy controls treated once a day with 0.15% dithranol in white petrolatum for 2 weeks. Skin biopsies were taken before and on days 2, 6, and 14 of the treatment. Lowry's microtechniques were used in conjunction with a bioluminescent system (bacterial luciferase) for enzymatic assays. The enzymic activity could be related to the type of keratinization present in the stratum corneum overlying the epidermal areas under study. In orthokeratotic areas from the controls, in noninvolved, and in treated involved skin the activity was low. In parakeratotic areas, as found in treated noninvolved and in involved psoriatic skin, the enzymic activity was increased to a level at least twice that found in orthokeratosis. Since ICDH(NAD) activity reflects an aspect of mitochondrial function, the results suggest that mitochondrial activity may be important in control of keratinization.

Topics: Adolescent; Adult; Anthracenes; Anthralin; Humans; Isocitrate Dehydrogenase; Keratins; Keratosis; Middle Aged; Mitochondria; NAD; Parakeratosis; Psoriasis; Skin

The ultrastructure of epidermis pretreated with 0,1 N sofium hydroxide, ether-ethanol mixture and 0,5 M sodium thioglycolate was studied. Sodium hydroxide dissolves the keratohyalin granules and the inter fibrillar material in the keratin. It has no effect on the epidermal and keratin fibers and on the marginal dense band of the horny cells. It "produces" a keratin pattern in the psoriatic horny layer. After sodium thioglycolate treatment the keratohyalin disappears, the epidermal and keratin fibers show a periodicity and the cementing material in the horny cells is more opaque. After lipid extraction the structure of keratohyalin becomes inhomogenous. The effect is somewhat similar to that of sodium thioglycolate but there is no periodicity in the horny layer. The intercellular contact layers of desmosomes as well as the Selby-Odland bodies disappear after each kind of treatment, The different effect of sodium thioglycolate on the horny layer and on the keratohyalin implies that the material of the keratohyalin and of the cementing substance are not identical.

Topics: Cytoplasmic Granules; Desmosomes; Humans; Keratins; Lipids; Microscopy, Electron; Psoriasis; Skin; Solvents

Twelve phenolic fractions of creosote and anthracene oils derived from a high temperature tar were applied in an ointment base to mouse tail skin. After treatment with the higher boiling acids, formerly parakeratotic scale areas underwent granular layer induction and 'basket-weave' keratin was produced. Changes in distribution of acid phosphatase and in horny layer fluorescence were consistent with the conversion to an orthokeratotic state. It is suggested that some of these phenols may be of value in the treatment of chronic psoriasis.

Topics: Acid Phosphatase; Animals; Anthracenes; Coal Tar; Creosote; Keratins; Mice; Parakeratosis; Phenols; Psoriasis; Skin

Topics: Abscess; Humans; Keratins; Leukocytes; Microscopy, Fluorescence; Psoriasis; Skin; Staining and Labeling; Suppuration; Sweat Glands

Topics: Adult; Biopsy; Cytoplasm; Cytoplasmic Granules; Histocytochemistry; Humans; Hyalin; Keratins; Male; Microscopy, Electron; Middle Aged; Psoriasis; Skin

Topics: Arthrodermataceae; Female; Germany, West; Humans; Keratins; Male; Nails; Onychomycosis; Psoriasis; Tinea Pedis

Topics: Bucladesine; Cells, Cultured; Cyclic AMP; Epinephrine; Epithelial Cells; Epithelium; Fluorides; Histamine; Humans; In Vitro Techniques; Isoproterenol; Keratins; Mitosis; Psoriasis; Theophylline

Topics: Animals; Cell Division; Connective Tissue; Disease Models, Animal; Epithelial Cells; Humans; Keratins; Melanocytes; Pigmentation Disorders; Psoriasis; Skin; Skin Diseases; Skin Neoplasms; Vitiligo

Topics: Biopsy; Humans; Keratins; Microscopy, Electron; Psoriasis; Skin

Topics: Amino Acids; Chromatography, Thin Layer; Electrophoresis, Cellulose Acetate; Humans; Hydrogen-Ion Concentration; Keratins; Proteins; Psoriasis; Skin; Solubility

Topics: Child; Child, Preschool; Diagnosis, Differential; Female; Hair; Humans; Keratins; Lice Infestations; Psoriasis; Skin Diseases

Topics: Autoradiography; Biopsy; Cell Differentiation; Computers; DNA; Female; Germ Cells; Humans; Keratins; Male; Middle Aged; Mitosis; Psoriasis; Skin; Thymidine; Time Factors; Tritium

Topics: Adolescent; Biopsy; Cytoplasm; Densitometry; Epithelial Cells; Epithelium; Humans; Keratins; Male; Methods; Microscopy, Electron; Microscopy, Electron, Scanning; Psoriasis; Skin

Topics: Animals; Antibody Formation; Cattle; Electrophoresis, Polyacrylamide Gel; Humans; Hydrogen-Ion Concentration; Ichthyosis; Immunodiffusion; Immunoelectrophoresis; Keratins; Mammals; Psoriasis; Rabbits; Rats; Skin

Topics: Animals; Animals, Newborn; Cell Division; Cells, Cultured; Colchicine; Cyclic AMP; Fibroblasts; Growth Inhibitors; Guinea Pigs; In Vitro Techniques; Keratins; Microscopy, Phase-Contrast; Psoriasis; Rats; Skin

Topics: Adolescent; Biopsy; Cell Nucleus; Cytoplasmic Granules; Female; Golgi Apparatus; Humans; Inclusion Bodies; Keratins; Keratosis; Langerhans Cells; Lipids; Lysosomes; Macrophages; Microscopy; Microscopy, Electron; Neutrophils; Organoids; Phagocytosis; Psoriasis; Skin

Topics: Cell Membrane Permeability; Humans; Keratins; Keratosis; Microscopy, Electron, Scanning; Psoriasis; Skin

Topics: Animals; Cattle; Electrophoresis, Starch Gel; Humans; Ichthyosis; Keratins; Peptide Biosynthesis; Protein Biosynthesis; Proteins; Psoriasis; Skin; Species Specificity; Structure-Activity Relationship; X-Ray Diffraction

Topics: Animals; Autoradiography; Cell Division; DNA Replication; Histidine; Humans; Keratins; Molecular Biology; Nucleic Acids; Protein Biosynthesis; Psoriasis; Rats; RNA; RNA, Messenger; Skin; Tritium

Topics: Amino Acids; Autoanalysis; Cadaver; Callosities; Chemical Precipitation; Chromatography, Gel; Dextrans; Electrophoresis, Polyacrylamide Gel; Humans; Hydrogen-Ion Concentration; Ichthyosis; Keratins; Protein Denaturation; Psoriasis; Skin; Tissue Extracts

Topics: Adult; Callosities; Female; Hair; Histocytochemistry; Humans; Infant; Intercellular Junctions; Keratins; Male; Methods; Microscopy, Electron; Nails; Osmium; Psoriasis; Skin; Staining and Labeling; Thioglycolates

Topics: Adolescent; Adult; Aged; Biological Assay; Erythrocytes; Female; Folic Acid; Folic Acid Deficiency; Humans; Keratins; Male; Middle Aged; Psoriasis; Skin; Tritium

Topics: Basement Membrane; Carcinoma; Dermatology; Diagnosis, Differential; Esterases; Fluorescent Antibody Technique; Glycogen; Histocytochemistry; Humans; Keratins; Keratoacanthoma; Keratosis; Lupus Erythematosus, Discoid; Mast-Cell Sarcoma; Psoriasis; Skin; Skin Diseases; Skin Neoplasms; Sweat Glands; Urticaria

Topics: Acne Vulgaris; Animals; Basement Membrane; Female; Humans; Injections, Intramuscular; Keratins; Male; Microscopy, Fluorescence; Psoriasis; Rats; Rosacea; Sebaceous Glands; Skin; Tail; Tetracycline

Topics: Animals; Cytoplasmic Granules; Histocytochemistry; Humans; Keratins; Marsupialia; Membranes; Microscopy, Electron; Phospholipids; Psoriasis; Skin

Topics: Carcinoma, Basal Cell; Humans; Ichthyosis; Keratins; Keratosis; Microscopy, Electron; Mitochondria; Psoriasis; Skin Neoplasms

Topics: Adolescent; Adult; Eczema; Female; Humans; Infrared Rays; Keratins; Male; Middle Aged; Psoriasis; Skin; Spectrophotometry

Topics: DNA; Humans; Ichthyosis; In Vitro Techniques; Keratins; Psoriasis; Ribosomes; RNA; Skin

Topics: Eczema; Esterases; Humans; Keratins; Phosphoric Monoester Hydrolases; Psoriasis; Skin; Staining and Labeling

Topics: Eczema; Electrophoresis; Esterases; Humans; Ichthyosis; Isoenzymes; Keratins; Psoriasis; Skin

Topics: Acid Phosphatase; Histocytochemistry; Humans; Keratins; Microscopy, Electron; Psoriasis; Skin

Topics: Alkaline Phosphatase; Antitoxins; Clostridium; Culture Media; Histocytochemistry; Histological Techniques; Keratins; Lipid Metabolism; Mice; Phospholipases; Phospholipids; Proteins; Psoriasis; Skin; Toxins, Biological

Topics: Biochemical Phenomena; Biochemistry; Callosities; DNA; Electrophoresis; Hair; Histocytochemistry; Humans; Keratins; Nails; Nitrogen; Proteins; Psoriasis; Skin; Sulfhydryl Compounds

Topics: Adolescent; Aprotinin; Chemical Phenomena; Chemistry; Child; Dermatology; Enzyme Inhibitors; Geriatrics; Humans; Isoflurophate; Kallikreins; Keratins; Psoriasis; Research Design; Tissue Culture Techniques; Trypsin

Topics: Absorption; Baths; Emulsions; Humans; Keratins; Mineral Oil; Oils; Psoriasis; Skin Diseases

Topics: Adolescent; Child; Drug Therapy; Geriatrics; Griseofulvin; Keratins; Leg Ulcer; Neurodermatitis; Pharmacology; Psoriasis; Tinea

Topics: Biomedical Research; Chromatography; Dermatitis, Exfoliative; Fractures, Bone; Hexosamines; Histocytochemistry; Humans; Keratins; Lipids; Psoriasis; Scalp; Skin

Topics: Biochemical Phenomena; Biochemistry; Cholesterol; Eczema; Humans; Keratins; Lipid Metabolism; Palmitic Acid; Psoriasis; Radiometry

Topics: Biopsy; Epidermis; Histocytochemistry; Humans; Keratins; Metabolism; Methionine; Psoriasis; Radiometry; Skin; Sulfates; Sulfur; Sulfur Isotopes

Topics: Alopecia; Alopecia Areata; Dermatitis; Dermatitis, Atopic; Dermatitis, Seborrheic; Dermatology; Humans; Keratins; Mercury; Nails; Ointments; Pigmentation; Psoriasis

Topics: Electrons; Epidermis; Keratins; Microscopy; Microscopy, Electron; Parakeratosis; Psoriasis; Skin

Topics: Electrons; Epidermis; Keratins; Microscopy; Microscopy, Electron; Parakeratosis; Psoriasis

Topics: Callosities; Epidermis; Humans; Keratins; Phosphates; Psoriasis; Skin; Water

Topics: Epidermis; Humans; Ichthyosis; Keratins; Psoriasis; Skin; X-Ray Diffraction

Topics: Acetates; Callosities; Hair; Keratins; Keratins, Type II; Nails; Psoriasis; Sulfhydryl Compounds; Thioglycolates

Topics: Cell Differentiation; Humans; Keratins; Psoriasis; Skin Physiological Phenomena