bromochloroacetic-acid and Herpes-Simplex

Topics: Age Factors; Carcinoma, Squamous Cell; Cell Membrane; Cell Nucleus; Cervix Mucus; Cervix Uteri; Chromatin; Cytoplasm; Female; Herpes Simplex; Humans; Hyperplasia; Inflammation; Keratins; Metaplasia; Pregnancy; Trichomonas Infections

Keratin is a strong, fibrous protein that coats the skin, male genitalia and some tissues in the mouth and female genitalia. Keratin protects the host by providing a barrier against primary herpes simplex virus infection. We postulate that keratin may also hinder the host by protecting the virus from effective immune responses such as those present on mucosal surfaces, enabling recurrent lesions to preferentially develop within keratinized tissues at mucocutaneous junctions. The natural history of infection supports the importance of preventing infection in keratinized tissues for developing an effective herpes simplex virus vaccine, since potent immunity on mucosal surfaces may be inadequate to protect keratinized tissues.

Topics: Female; Herpes Simplex; Herpes Simplex Virus Vaccines; Humans; Immunity, Mucosal; Keratins; Male; Recurrence; Simplexvirus; Viral Vaccines

The severe acute respiratory syndrome (SARS) pandemic in Toronto resulted in a large number of autopsies on its victims. We describe the pulmonary pathology of patients who died in the 2003 Toronto outbreak. Autopsy material from the lungs of 20 patients who died between March and July 2003 were characterized by histology, molecular biology, and immunohistochemistry for cytokeratins, thyroid transcription factor-1, CD68, Epstein-Barr virus, cytomegalovirus, and human herpes simplex viruses. Matched controls were obtained from patients who died of other causes over the same interval. The mean duration of illness was 27 days (range 5-108 days). Post-mortem lung tissues from 19 of 20 patients with probable SARS were positive for SARS-associated coronavirus by RT-PCR. Histologically, all patients showed varying degrees of exudative and proliferative phase acute lung injury, evidenced in conventional and immunohistochemical stains by edema, inflammatory infiltrate, pneumocyte hyperplasia, fibrinous exudates, and organization. Eight of 20 patients showed predominantly a diffuse alveolar damage pattern of acute lung injury, six showed predominantly an acute fibrinous and organizing pneumonia pattern, and the remainder showed an admixture of the two patterns. Squamous metaplasia and scattered multinucleate giant cells were present in most cases. Vascular fibrin thrombi were a common finding and were often associated with pulmonary infarcts. Special stains demonstrated vascular endothelial damage of both small- and mid-sized pulmonary vessels. Two cases were complicated by invasive fungal disease consistent with Aspergillosis, and another by coinfection with cytomegalovirus. Our findings indicate that the lungs of patients who die of SARS are almost always positive for the SARS-associated coronavirus by RT-PCR, and may show features of both diffuse alveolar damage and acute fibrinous and organizing pneumonia patterns of acute injury. Cases of SARS may be complicated by coexistent infections and therapy-related lung injury.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Aspergillosis; Aspergillus; Autopsy; Canada; Cytomegalovirus; Cytomegalovirus Infections; Female; Herpes Simplex; Humans; Immunohistochemistry; Keratins; Lung; Lung Diseases, Fungal; Male; Middle Aged; Nuclear Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral; Severe Acute Respiratory Syndrome; Severe acute respiratory syndrome-related coronavirus; Simplexvirus; Thyroid Nuclear Factor 1; Transcription Factors; Viral Matrix Proteins

In human recurrent herpetic lesions epidermal keratinocytes are induced to express HLA class II (DR) antigens. Keratinocytes derived from human split skin and cultured in vitro were induced to express HLA-DR but not -DQ antigens with IFN gamma preparations. These stimulated keratinocytes presented herpes simplex antigen directly to autologous blood-derived T lymphocytes in four of four subjects (stimulation indices: 1.5-2.7), suggesting that keratinocytes may have an accessory herpes simplex virus (HSV) antigen-presenting role in addition to the Langerhans cells and macrophages in herpetic skin lesions. Blood mononuclear cells from eight herpes simplex seropositive subjects which were activated in vitro by HSV antigen for 6 d showed cytotoxicity specific for HSV in infected autologous keratinocytes. This was significantly increased by prestimulation with IFN gamma (51-56% to 83-85%). In four of eight patients some cytotoxicity also occurred against uninfected, IFN gamma-stimulated keratinocytes. Lymphocyte subset analysis showed that cytotoxicity against HSV-infected, IFN gamma-stimulated keratinocyte targets was mediated by both CD3+ T lymphocytes and Leu 11b+ natural killer cells. T lymphocyte cytotoxicity was mediated by both CD4+ and CD8+ T lymphocytes, suggesting a cytotoxic role for the activated CD4+ lymphocytes that initially predominate in herpetic lesions.

Topics: Antigens, Differentiation, T-Lymphocyte; Antigens, Viral; CD3 Complex; CD8 Antigens; Cytotoxicity, Immunologic; Epidermis; Herpes Simplex; HLA-DR Antigens; Humans; Interferon-gamma; Keratins; Receptors, Antigen, T-Cell; Simplexvirus; T-Lymphocytes, Cytotoxic

Herpes simplex virus (HSV) specific immune mediated cytotoxicity may be involved in control of HSV infections and in the tissue damage induced by HSV infection or HSV related skin disease such as herpes associated erythema multiforme. Developing an in vitro model to study this process has proved difficult due to the lack of an appropriate target cell that will express HSV antigens but is not simultaneously subject to viral induced cell death. The purpose of this study was to develop a model in which keratinocytes express cell surface HSV specific antigens but at the same time are protected from death due to viral infection. We found that keratinocytes infected with HSV in the presence of acyclovir (ACV) expressed such antigens yet remained viable for a period of time after the onset of antigen expression such that cytotoxicity studies could be successfully performed. Rabbit skin cells, a transformed keratinocyte line, or second passage human neonatal foreskin keratinocytes were grown in culture medium with or without 200 microM ACV and were infected with HSV. Examination by direct immunofluorescence with anti-HSV antibody revealed uniform HSV antigen expression by cells both with and without ACV by 8 h after infection. Cells infected without ACV exhibited marked structural abnormalities including formation of multinucleated giant cells, while cells with ACV showed fewer such changes throughout a 24-h period. An Ethidium Bromide-Acridine Orange cytotoxicity assay demonstrated significant increases in the cytotoxicity of infected cells not protected by ACV compared to that of cells with ACV (p less than .001). This in vitro model should prove useful in the investigation of HSV specific immune mediated cytotoxicity.

Topics: Acridine Orange; Acyclovir; Animals; Antigens, Viral; Cells, Cultured; Cytotoxicity Tests, Immunologic; Cytotoxicity, Immunologic; Epidermal Cells; Epidermis; Epitopes; Ethidium; Herpes Simplex; Keratins; Simplexvirus

beta Interferon (IFN) was demonstrated by specific, sequential antibody-neutralization assays of vesicle fluids from patients with recurrent skin lesions due to herpes simplex virus. To determine the origin of this antiviral activity, we cultured keratinocytes from normal facial skin and infected them with three strains of herpes simplex virus. Keratinocyte cultures then developed characteristic cytopathic changes, and antiviral activity was found in culture supernatant media. All such activity from these supernatants was neutralized with specific antiserum to IFN-beta but not with antiserum to IFN-alpha. No IFN-gamma was detectable by radioimmunoassay. Immunoperoxidase staining with antiserum to IFN-beta in five biopsy specimens from culture-proven, recurrent herpes simplex lesions showed positive staining of epidermal keratinocytes but not of dermal or infiltrating cells. Thus, the primary sources of IFN-beta in recurrent herpes lesion vesicles are the virus-infected keratinocytes.

Topics: Cells, Cultured; Cytopathogenic Effect, Viral; Epidermis; Herpes Labialis; Herpes Simplex; Humans; Interferon Type I; Keratins; Recurrence; Simplexvirus

The fate of microtubules and of vimentin or keratin containing intermediate filaments during infection with fusion or rounding producing strains of herpes simplex virus (HSV) was investigated. Microtubules polymerize early after fusion of cells. However, they do not reconstitute 6-7 hours post infection (p.i.) after release of a colcemid block. Keratin and vimentin are maintained around the original nucleus still inside of recruited cells in the polykaryocyte. Cells of fibroblastic and epithelial origin fuse. Inside of polykaryocytes keratin or vimentin containing fibers seem to polymerize. Keratin is to be found in invaginations in the nuclei surrounded by the inner layer of the nuclear membrane. Anti-keratin antibodies specifically label HSV envelopes located in the cytoplasm or outside of the cell. Controls of the procedure allowed to exclude labelling of HSV envelopes via gpE, which represents HSV induced Fc receptors. Late stages of infected cells contain thickened and condensed keratin fibers. Conversely, vimentin fibers late after infection appear to be evenly distributed and to be thin. Microtubules decay late after infection with rounding producing strains of HSV, whereas keratin and vimentin fibers are still present late after infection.

Topics: Animals; Cell Fusion; Cell Line; Cytopathogenic Effect, Viral; Cytoskeleton; Epithelium; Fibroblasts; Herpes Simplex; Intermediate Filaments; Keratins; Microtubules; Simplexvirus; Vimentin

Topics: Cytotoxicity Tests, Immunologic; Epidermis; Granulocytes; Herpes Simplex; Herpes Zoster; Humans; Keratins; Lymphocytes; Rosette Formation; Skin Diseases; Skin Diseases, Vesiculobullous

Topics: Birth Injuries; Blister; Drug Eruptions; Epidermolysis Bullosa; Erythema; Female; Herpes Simplex; Humans; Infant; Infant, Newborn; Infant, Newborn, Diseases; Keratins; Listeria monocytogenes; Lupus Erythematosus, Systemic; Male; Maternal-Fetal Exchange; Miliaria; Nevus; Pregnancy; Pregnancy Complications; Rubella; Skin Diseases; Skin Diseases, Infectious; Stevens-Johnson Syndrome; Sucking Behavior; Syphilis, Congenital; Urticaria Pigmentosa