bromochloroacetic-acid and Gonorrhea

To determine the characteristics of regenerated epithelial cells after severe gonococcal infection after corneal perforation.. Pathological tissue was obtained from the cornea at the time of surgery. Hematoxylin and eosin staining and immunohistochemical analysis were performed for cytoskeletal keratins (K12, K13, and K15), basement membrane and junctional markers (laminin 5, ZO-1 and Desmoplakin), and proliferative and mesenchymal markers (Ki67, α-SMA, and vimentin).. A 42-year-old patient with severe gonococcal keratoconjunctivitis rapidly progressed to corneal perforation during administration of intensive topical and systemic antibiotics. After conservative treatment, the perforation healed and 5- × 3-mm corneal ectasia occurred with localized iris attachment. Complete closure of the cornea was confirmed by a negative Seidel test. After lamellar keratoplasty to improve corneal integrity and to prevent secondary glaucoma, the pathological tissue revealed a poorly organized epithelial layer at the regenerated ectatic area. The regenerated epithelial cells clearly expressed K12, ZO-1, and Desmoplakin with underlying laminin 5 (+) basement membrane. K15 and Ki67 expressions were observed predominantly at the limbal area but not in the regenerated area. α-SMA and vimentin were sporadically expressed in the underlying connective tissue.. We speculate that the process of epithelial wound healing at the site of corneal perforation was responsible for migration of the surrounding epithelial cells. Although the regenerated cells expressed several cytokeratins and junctional markers, they remained disorganized and fragile.

Topics: Actins; Adult; Biomarkers; Cell Movement; Corneal Perforation; Corneal Transplantation; Epithelial Cells; Epithelium, Corneal; Eye Infections, Bacterial; Gonorrhea; Humans; Immunohistochemistry; Keratins; Ki-67 Antigen; Male; Neisseria gonorrhoeae; Phenotype; Regeneration; Tight Junction Proteins; Vimentin; Wound Healing

The primary human urethral epithelial cells developed by our laboratory have been immortalized by transduction with a retroviral vector expressing the human papillomavirus E6E7 oncogenes. Analysis of telomerase expression and comparison to that in primary cells revealed detectable levels in the transduced human urethral epithelial cells. Immortalized urethral cells could be passaged over 20 times. Immunofluorescence microscopy studies showed that the immortalized cells were phenotypically similar and responded to gonococcal infection similarly to primary cells. Specifically, positive cytokeratin staining showed that the immortalized cells are keratinocytes; cell surface levels of human asialoglycoprotein receptor increase following gonococcal infection, and, like the primary cells, the immortalized urethral epithelial cells are CD14 negative. Using enzyme-linked immunosorbent assay, we found that interleukin-6 (IL-6) and IL-8 levels in primary urethral epithelial cell supernatants increase after challenge with N. gonorrhoeae. Likewise, the immortalized urethral epithelial cells produced higher levels of IL-6 and IL-8 cytokines in response to gonococcal infection. Cells challenged with a gonococcal lipid A msbB mutant produced reduced IL-6 and IL-8 levels when compared to the parent strain. Additionally, these data suggest that the 1291 msbB lipooligosaccharide may suppress cytokine induction.

Topics: Asialoglycoprotein Receptor; Cell Transformation, Viral; Cells, Cultured; Cytokines; Epithelial Cells; Genes, Viral; Genetic Vectors; Gonorrhea; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Karyotyping; Keratins; Lipopolysaccharide Receptors; Models, Biological; Neisseria gonorrhoeae; Oncogene Proteins, Viral; Papillomavirus E7 Proteins; Phenotype; Receptors, Cell Surface; Retroviridae; Urethra