Abstract
A tissue culture model has been developed for studying the ability of Neisseria gonorrhoeae to invade eucaryotic cells. The cell line HecIB, a human adenocarcinoma endometrial cell line, was found to support gonococcal invasion. The bactericidal antibiotic gentamicin was used to kill those bacteria that had not entered the HecIB cells, allowing us to quantitate internalized bacteria. Kinetic studies showed an increase in the titer of gentamicin-protected gonococci at 4 h postinfection followed by a decrease; a second increase occurred after 6 h. The state of piliation did not affect the degree of invasion when the bacteria were spun down onto the monolayer. Gonococcal invasion was inhibited when the HecIB cells were preincubated with cytochalasin D before bacterial infection. N. lactamica was used as a negative control. No internalized N. lactamica cells were observed by electron microscopy. Electron microscopy documented the intracellular location of the gonococci in HecIB cells and the eventual destruction of the invaded HecIB cells. After 24 h, clusters of gonococci encased in a matrix of cell debris were observed.
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Selected References
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- Bessen D., Gotschlich E. C. Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II. Infect Immun. 1986 Oct;54(1):154–160. doi: 10.1128/iai.54.1.154-160.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Black W. J., Cannon J. G. Cloning of the gene for the common pathogenic Neisseria H.8 antigen from Neisseria gonorrhoeae. Infect Immun. 1985 Jan;47(1):322–325. doi: 10.1128/iai.47.1.322-325.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukholm G. Effect of cytochalasin B and dihydrocytochalasin B on invasiveness of entero-invasive bacteria in HEp-2 cell cultures. Acta Pathol Microbiol Immunol Scand B. 1984 Jun;92(3):145–149. doi: 10.1111/j.1699-0463.1984.tb02809.x. [DOI] [PubMed] [Google Scholar]
- Gregg C. R., Melly M. A., Hellerqvist C. G., Coniglio J. G., McGee Z. A. Toxic activity of purified lipopolysaccharide of Neisseria gonorrhoeae for human fallopian tube mucosa. J Infect Dis. 1981 Mar;143(3):432–439. doi: 10.1093/infdis/143.3.432. [DOI] [PubMed] [Google Scholar]
- Hale T. L., Morris R. E., Bonventre P. F. Shigella infection of henle intestinal epithelial cells: role of the host cell. Infect Immun. 1979 Jun;24(3):887–894. doi: 10.1128/iai.24.3.887-894.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hodinka R. L., Wyrick P. B. Ultrastructural study of mode of entry of Chlamydia psittaci into L-929 cells. Infect Immun. 1986 Dec;54(3):855–863. doi: 10.1128/iai.54.3.855-863.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Isberg R. R., Falkow S. A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature. 1985 Sep 19;317(6034):262–264. doi: 10.1038/317262a0. [DOI] [PubMed] [Google Scholar]
- Kihlström E., Nilsson L. Endocytosis of Salmonella typhimurium 395 MS and MR10 by HeLa cells. Acta Pathol Microbiol Scand B. 1977 Oct;85B(5):322–328. doi: 10.1111/j.1699-0463.1977.tb01982.x. [DOI] [PubMed] [Google Scholar]
- Koomey J. M., Gill R. E., Falkow S. Genetic and biochemical analysis of gonococcal IgA1 protease: cloning in Escherichia coli and construction of mutants of gonococci that fail to produce the activity. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7881–7885. doi: 10.1073/pnas.79.24.7881. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGee Z. A., Johnson A. P., Taylor-Robinson D. Human fallopian tubes in organ culture: preparation, maintenance, and quantitation of damage by pathogenic microorganisms. Infect Immun. 1976 Feb;13(2):608–618. doi: 10.1128/iai.13.2.608-618.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McGee Z. A., Stephens D. S., Hoffman L. H., Schlech W. F., 3rd, Horn R. G. Mechanisms of mucosal invasion by pathogenic Neisseria. Rev Infect Dis. 1983 Sep-Oct;5 (Suppl 4):S708–S714. doi: 10.1093/clinids/5.supplement_4.s708. [DOI] [PubMed] [Google Scholar]
- Melly M. A., Gregg C. R., McGee Z. A. Studies of toxicity of Neisseria gonorrhoeae for human fallopian tube mucosa. J Infect Dis. 1981 Mar;143(3):423–431. doi: 10.1093/infdis/143.3.423. [DOI] [PubMed] [Google Scholar]
- Ota F., Pontefract R., Ashton F. E., Diena B. B. Studies on gonococcal infection. II. Attachment and fate of gonococci in tissue-culture cells. Can J Microbiol. 1975 Nov;21(11):1698–1704. doi: 10.1139/m75-249. [DOI] [PubMed] [Google Scholar]
- Sansonetti P. J., Kopecko D. J., Formal S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. doi: 10.1128/iai.35.3.852-860.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Small P. L., Isberg R. R., Falkow S. Comparison of the ability of enteroinvasive Escherichia coli, Salmonella typhimurium, Yersinia pseudotuberculosis, and Yersinia enterocolitica to enter and replicate within HEp-2 cells. Infect Immun. 1987 Jul;55(7):1674–1679. doi: 10.1128/iai.55.7.1674-1679.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tyeryar F. J., Jr, Quan A. L., Rene A. A., Weiss E. Phase transition of gonococci in mammalian cell cultures. Infect Immun. 1974 Dec;10(6):1401–1411. doi: 10.1128/iai.10.6.1401-1411.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaudaux P., Waldvogel F. A. Gentamicin antibacterial activity in the presence of human polymorphonuclear leukocytes. Antimicrob Agents Chemother. 1979 Dec;16(6):743–749. doi: 10.1128/aac.16.6.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waitkins S. A., Flynn J. Intracellular growth and type variation of Neisseria gonorrhoeae in tissue cell-cultures. J Med Microbiol. 1973 Aug;6(3):399–403. doi: 10.1099/00222615-6-3-399. [DOI] [PubMed] [Google Scholar]
- Ward M. E., Watt P. J. Adherence of Neisseria gonorrhoeae to urethral mucosal cells: an electron-microscopic study of human gonorrhea. J Infect Dis. 1972 Dec;126(6):601–605. doi: 10.1093/infdis/126.6.601. [DOI] [PubMed] [Google Scholar]
- Ward M. E., Watt P. J., Robertson J. N. The human fallopian tube: a laboratory model for gonococcal infection. J Infect Dis. 1974 Jun;129(6):650–659. doi: 10.1093/infdis/129.6.650. [DOI] [PubMed] [Google Scholar]