Affiliation:
1. Department of Microbiology, The University of Iowa, Iowa City, Iowa 52242
Abstract
ABSTRACT
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.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Immunology,Microbiology,Parasitology
Reference31 articles.
1. Alberts B. D. Bray J. Lewis M. Raff K. Roberts and J. D. Watson. 1989. Molecular biology of the cell p. 752-761. Garland Publishing Inc. New York N.Y.
2. Apicella, M. A., M. R. Ketterer, F. K. N. Lee, D. Zhou, P. A. Rice, and M. S. Blake. 1996. The pathogenesis of gonococcal urethritis in men: confocal and immunoelectron microscopic analysis of urethral exudates from men infected with Neisseria gonorrhoeae. J. Infect. Dis.173:636-646.
3. Baccam, M., and G. A. Bishop. 1999. Membrane-bound CD154, but not CD40-specific antibody, mediates NF-κB-independent IL-6 production in B cells. Eur. J. Immunol.29:3855-3866.
4. A primer on cytokines: sources, receptors, effects, and inducers
5. Neisseria gonorrhoeae
Induces Focal Polymerization of Actin in Primary Human Urethral Epithelium
Cited by
53 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献