bromochloroacetic-acid and Streptococcal-Infections

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

Early events leading to intrauterine infection remain poorly defined, but may hold the key to preventing preterm delivery. To determine molecular pathways within fetal membranes (chorioamnion) associated with early choriodecidual infection that may progress to preterm premature rupture of membranes (PPROM), we examined the effects of a Group B Streptococcus (GBS) choriodecidual infection on chorioamnion in a nonhuman primate model. Ten chronically catheterized pregnant monkeys (Macaca nemestrina) at 118-125 days gestation (term = 172 days) received choriodecidual inoculation of either GBS (n = 5) or saline (n = 5). Cesarean section was performed in the first week after GBS or saline inoculation. RNA extracted from chorioamnion (inoculation site) was profiled by microarray. Single gene, Gene Set, and Ingenuity Pathway Analysis results were validated using qRT-PCR (chorioamnion), Luminex (amniotic fluid, AF), immunohistochemistry, and transmission electron microscopy (TEM). Despite uterine quiescence in most cases, significant elevations of AF cytokines (TNF-α, IL-8, IL-1β, IL-6) were detected in GBS versus controls (p<0.05). Choriodecidual infection resolved by the time of cesarean section in 3 of 5 cases and GBS was undetectable by culture and PCR in the AF. A total of 331 genes were differentially expressed (>2-fold change, p<0.05). Remarkably, GBS exposure was associated with significantly downregulated expression of multiple cytokeratin (CK) and other cytoskeletal genes critical for maintenance of tissue tensile strength. Immunofluorescence revealed highly significant changes in the CK network within amniocytes with dense CK aggregates and retraction from the cell periphery (all p = 0.006). In human pregnancies affected by PPROM, there was further evidence of CK network retraction with significantly shorter amniocyte foot processes (p = 0.002). These results suggest early choriodecidual infection results in decreased cellular membrane integrity and tensile strength via dysfunction of CK networks. Downregulation of CK expression and perturbations in the amniotic epithelial cell intermediate filament network occur after GBS choriodecidual infection, which may contribute to PPROM.

Topics: Amnion; Animals; Chorion; Disease Models, Animal; Female; Fetal Membranes, Premature Rupture; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Keratins; Macaca nemestrina; Microscopy, Confocal; Microscopy, Electron, Transmission; Oligonucleotide Array Sequence Analysis; Pregnancy; Prenatal Exposure Delayed Effects; Reverse Transcriptase Polymerase Chain Reaction; Streptococcal Infections; Streptococcus agalactiae; Transcriptome

In contrast to wild-type mice, genetically engineered Mucin1 (Muc1) null animals display a marked propensity for development of blepharitis and conjunctivitis. Molecular approaches confirmed the presence of Muc1 mRNA and protein in the conjunctival tissue of wild-type mice and identified the bacterial species in Muc1 null symptomatic mice.. Muc1 null animals housed in a conventional facility were examined for visually apparent inflammation of the eye and surrounding tissue. Blood taken from overtly affected animals was assayed for antibodies to common murine viral agents. Swabs of infected eyes and whole eye preparations were used to detect and speciate bacterial pathogens. Frozen sections of whole eye, lid margin, and Harderian gland were immunostained with antibodies to Muc1 and cytokeratin 14, both epithelial cell markers. Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) were performed on RNA isolated from conjunctiva and Harderian gland of wild-type mice to compare relative levels of transcript.. Student's unpaired t-test performed on the eye inflammation frequency of Muc1 null mice confirmed a statistical significance (P < 0.01) when compared to wild-type background animals housed in the same room. Analysis of blood samples from affected Muc1 null animals detected no common murine viral pathogens. Bacterial analysis of conjunctival swabs and whole eye preparations demonstrated the presence of coagulase-negative Staphylococcus, Streptococcus type alpha, and Corynebacterium group G2. Muc1 antibody staining of wild-type sections revealed the presence of Muc1 on conjunctival goblet and non-goblet cells and on the epithelium of the Harderian gland. Serial sections stained with cytokeratin 14 antibody confirmed the epithelial nature of cells expressing the Muc1 protein. RNA from conjunctiva and Harderian gland subjected to RT-PCR and northern blot analysis showed an abundance of Muc1 transcript in these tissues.. Muc1 mRNA and protein are present in murine conjunctival and Harderian gland epithelia. Animals lacking Muc1 mRNA and protein are predisposed to developing eye inflammation when compared to wild-type animals with an intact Muc1 gene. Muc1 appears to play a critical protective role at the ocular surface, presumably by acting as a barrier to infection by certain bacterial strains.

Topics: Animals; Blepharitis; Conjunctiva; Conjunctivitis, Bacterial; Corynebacterium Infections; DNA Primers; Female; Fluorescent Antibody Technique, Indirect; Harderian Gland; Keratins; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Mucin-1; Polymerase Chain Reaction; RNA, Messenger; Staphylococcal Infections; Streptococcal Infections

The distribution of collagen types I, III, and IV and of laminin, fibronectin, and keratin was studied in otitis media experimentally induced by Streptococcus pneumoniae in the chinchilla. The expression of interstitial collagen types I and III and of fibronectin was increased in the subepithelial space that was thickened by inflammation in the acute period of infection. The expression of collagen type IV in the subepithelial space could be seen in the early period. The epithelial cells in the middle ear changed from flat cuboidal to pseudostratified columnar in pneumococcus-inoculated ears, and the number of keratin-positive epithelial cells in the middle ear increased remarkably after infection. These results indicate that changes in epithelial cell differentiation effected by the extracellular matrix correlate with changes in expression of keratin. It is proposed that the extracellular matrix may contribute to tissue repair in the middle ear after bacterial infection by interfering with cell proliferation of epithelial cells and fibroblasts.

Topics: Animals; Cell Differentiation; Chinchilla; Collagen; Ear, Middle; Epithelial Cells; Extracellular Matrix; Extracellular Matrix Proteins; Fibronectins; Keratins; Mucous Membrane; Otitis Media; Streptococcal Infections; Streptococcus pneumoniae

Besides group A (GAS), Lancefield group C beta-haemolytic streptococci (GCS) have been implicated as a causative agent in outbreaks of purulent pharyngitis. In this study we have investigated a class CI M protein of a Streptococcus dysgalactiae1:256, revealed that 26% of these sera showed serological cross-reactivity between a 68-kDa cartilage protein and the N-terminal part of MC. Only 8% of the sera of healthy patients showed this property. In additional, MC also cross-reacted with antibodies recognising epidermal keratins. The cross-reacting 68-kDa protein from cartilage was different from human serum albumin, but was recognised with anti-vimentin immune serum. The MC was cloned and the gene sequenced. By using PCR, recombinant gene fragments encoding characteristic peptide fragments of MC were expressed in Escherichia coli. The peptides were used to map the binding sites for plasma proteins and to locate the cross-reacting epitopes on the MC molecule. In consequence, sequence alignments revealed that MC shared homologous regions with vimentin and different keratins. Our data, obtained with MC, suggest that not only infections with GAS but also infections with GCS and possibly GGS (the latter species can also produce class CI M-like proteins) may be responsible for the formation of streptococcal-associated sequel diseases.

Topics: Amino Acid Sequence; Animals; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Blood Proteins; Blotting, Western; Carrier Proteins; Cartilage; Chick Embryo; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Epitopes; Humans; Immune Sera; Keratins; Molecular Sequence Data; Phagocytosis; Protein Binding; Rabbits; Sequence Homology, Amino Acid; Streptococcal Infections; Streptococcus; Virulence; Wounds and Injuries

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

Physical characteristics of the bovine teat canal were examined for their influence on susceptibility to intramammary infection. All quarters of 18 cows were inoculated with 2 x 10(5) cfu Streptococcus agalactiae (Trial 1) and 20 cows with 10(5) cfu Str. uberis (Trial 2) 3-4 mm into the teat canal every 3 d for 12 d. Incidence of quarter infection was similar for both pathogens, 30/72 (42%) in Trial 1 and 32/80 (40%) in Trial 2. Logistic regression analysis showed that probability of infection by Str. agalactiae increased significantly with an increase in quarter peak flow rate (P < 0.05) whereas probability of infection increased for Str. uberis with a decrease in teat canal length (P < 0.05). A significantly higher (P < 0.001) incidence of infection by Str. uberis was observed in quarters that contained a low wet weight (< 1.8 mg) of removable keratin compared with those that contained > 1.8 mg keratin, but there was no correlation between weight of keratin and length of the teat canal. Infections by Str. uberis took significantly less (P < 0.05) time to show a rise in somatic cell count above 7.5 x 10(5) cells/ml than Str. agalactiae. The results provide evidence that these pathogens use different mechanisms to pass through the teat canal.

Topics: Animals; Cattle; Dairying; Disease Susceptibility; Female; Keratins; Kinetics; Lactation; Mammary Glands, Animal; Mastitis, Bovine; Regression Analysis; Streptococcal Infections; Streptococcus agalactiae

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

Influence of teat canal keratin on susceptibility to intramammary infection was investigated in lactating Jersey cows. In each of two replicate trials, keratin was removed from the left teats of 20 cows immediately before milking. Immediately after milking, all teats were exposed to bacterial challenge by immersion in a suspension of Streptococcus agalactiae (5 x 10(7) cfu/ml). Bacterial challenge was repeated after the next four milkings. Foremilk samples were obtained for 8 d after keratin removal to determine infection status. A mammary quarter was classified as infected based solely upon the bacteriological criteria outlined by the National Mastitis Council. The rate of infection in quarters from which keratin was removed was greater than that in control quarters. Infection rates were 26.3% for keratin-removed quarters and 8.3% for control quarters in trial 1 and 13.5 and 0%, respectively, in trial 2. When more stringent criteria (recovery of greater than 100 cfu of S. agalactiae/ml in three or more successive milk samples and a SCC of greater than 10(6)) were used to identify a subset of infections that were clearly intramammary, infection rates were 9.3% for keratin-removed quarters and 1.4% for control quarters. Thus, partial removal of keratin from the teat canal compromised the ability of the teat to prevent passage of bacterial pathogens from the external environment into the mammary gland.

Topics: Animals; Cattle; Disease Susceptibility; Female; Immunity, Innate; Keratins; Mammary Glands, Animal; Mastitis, Bovine; Streptococcal Infections; Streptococcus agalactiae

Teat canal keratin (n = 461) and mammary gland secretions (n = 370) were collected from 31 unbred and 85 primigravid Jersey heifers from one research and three commercial dairy herds. Of 97 heifers from which secretion samples were obtained, 96.9% had intramammary infections and 29% showed clinical symptoms. Seventy-five percent of quarters were infected. Staphylococcus aureus were isolated from 36 (37.1%) heifers and 55 (14.9%) quarters. One hundred and eight (93.1%) heifers and 326 (70.7%) quarters had teat canals colonized with mastitis pathogens. Staphylococcus aureus were isolated from teat canal keratin samples from 36 (31%) heifers and 57 (12.3%) quarters. The three most common species isolated from secretion and teat canal keratin samples were Staphylococcus chromogenes, Staphylococcus hyicus, and S. aureus. Secretions from infected (n = 240) and uninfected (n = 85) quarters had SCC of 13.6 X 10(6)/ml and 5.7 X 10(6)/ml. Macrophages were the most numerous cell type in secretions of infected and uninfected quarters. Quarters with teat canal colonization, but with no intramammary infections, exhibited higher SCC in secretion (9.3 X 10(6)/ml) than quarters without both teat canal colonizations and intramammary infections (4.9 X 10(6)/ml). Data indicated that intramammary infections and teat canal colonizations were more prevalent and SCC higher than previously realized in dairy heifers.

Topics: Animals; Cattle; Female; Keratins; Leukocyte Count; Lymphocytes; Macrophages; Mammary Glands, Animal; Mastitis, Bovine; Neutrophils; Pregnancy; Pregnancy Complications, Infectious; Prevalence; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus; Streptococcal Infections; Streptococcus