Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1977 Feb;15(2):560–567. doi: 10.1128/iai.15.2.560-567.1977

In vitro interaction of Neisseria gonorrhoeae type 1 and type 4 with tissue culture cells.

B R Brodeur, W M Johnson, K G Johnson, B B Diena
PMCID: PMC421404  PMID: 403139

Abstract

As a basis for studies of gonococcal pathogenicity, tissue culture cells were infected with type 1 or type 4 Neisseria gonorrhoeae to determine intracellular viability. A simple and objective means of measurement was devised, based on the uptake of tritiated protein and deoxyribonucleic acid precursors by cycloheximide-inhibited cells infected with gonococci. Cycloheximide was found to inhibit protein synthesis by over 97% tissue culture cells at a concentration of 100 microng/ml. In contrast, N, gonorrhoeae was found to be highly resistant to this antibiotic, and protein synthesis was unaffected by concentrations up to 1,000 microng/ml. Extracellular gonococci were eliminated by treatment with high concentrations of penicillin during cycloheximide inhibition and prior to the addition of radioisotope. Levels of protein and deoxyribonucleic acid synthesis by N. gonorrhoeae in the cycloheximide-treated cells were significantly higher in T1-infected cells (RE2, HeLa, or HEp-2) than in the corresponding T4-infected cells. No differences were observed in tissue cell susceptibility to gonococcal infection. Intracytoplasmic localization of N. gonorrhoeae was confirmed by electron microscopy.

Full text

PDF
560

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Calabi O. In-vitro interaction of Shigella flexneri with leukocytes and HeLa cells. J Infect Dis. 1970 Jul-Aug;122(1):1–9. doi: 10.1093/infdis/122.1-2.1. [DOI] [PubMed] [Google Scholar]
  2. Catlin B. W. Nutritional profiles of Neisseria gonorrhoeae, Neisseria meningitidis, and Neisseria lactamica in chemically defined media and the use of growth requirements for gonococcal typing. J Infect Dis. 1973 Aug;128(2):178–194. doi: 10.1093/infdis/128.2.178. [DOI] [PubMed] [Google Scholar]
  3. Dilworth J. A., Hendley J. O., Mandell G. L. Attachment and ingestion of gonococci human neutrophils. Infect Immun. 1975 Mar;11(3):512–516. doi: 10.1128/iai.11.3.512-516.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ellen R. P., Gibbons R. J. M protein-associated adherence of Streptococcus pyogenes to epithelial surfaces: prerequisite for virulence. Infect Immun. 1972 May;5(5):826–830. doi: 10.1128/iai.5.5.826-830.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Esquenazi V., Streitfeld M. M. Transformation of lymphocytes in gonorrhea before and after therapy. Infect Immun. 1973 Oct;8(4):503–509. doi: 10.1128/iai.8.4.503-509.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FRIEDKIN M., WOOD H. I. V. Utilization of thymidine-C14 by bone marrow cells and isolated thymus nuclei. J Biol Chem. 1956 Jun;220(2):639–651. [PubMed] [Google Scholar]
  7. Farzadegan H., Roth I. L. Scanning electron microscopy and freeze-etching of gonorrhoeal urethral exudate. Br J Vener Dis. 1975 Apr;51(2):83–91. doi: 10.1136/sti.51.2.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glynn A. A., Ward M. E. Nature and Heterogeneity of the Antigens of Neisseria gonorrhoeae Involved in the Serum Bactericidal Reaction. Infect Immun. 1970 Aug;2(2):162–168. doi: 10.1128/iai.2.2.162-168.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grimble A. S., McIllmurray M. B. Cell-mediated immune response in gonorrhoea. Br J Vener Dis. 1973 Oct;49(5):446–449. doi: 10.1136/sti.49.5.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hill J. M., Morse P. A., Jr, Gentry G. A. Metabolism of deoxycytidine, thymine, and deoxythymidine in the hamster. Cancer Res. 1975 May;35(5):1314–1319. [PubMed] [Google Scholar]
  11. James-Holmquest A. N., Swanson J., Buchanan T. M., Wende R. D., Williams R. P. Differential attachment by piliated and nonpiliated Neisseria gonorrhoeae to human sperm. Infect Immun. 1974 May;9(5):897–902. doi: 10.1128/iai.9.5.897-902.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KELLOGG D. S., Jr, PEACOCK W. L., Jr, DEACON W. E., BROWN L., PIRKLE D. I. NEISSERIA GONORRHOEAE. I. VIRULENCE GENETICALLY LINKED TO CLONAL VARIATION. J Bacteriol. 1963 Jun;85:1274–1279. doi: 10.1128/jb.85.6.1274-1279.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kraus S. J., Perkins G. H., Geller R. C. Lymphocyte transformation in repeated gonococcal urethritis. Infect Immun. 1970 Nov;2(5):655–658. doi: 10.1128/iai.2.5.655-658.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Novotny P., Short J. A., Walker P. D. An electron-microscope study of naturally occurring and cultured cells of Neisseria Gonorrhoeae. J Med Microbiol. 1975 Aug;8(3):413–427. doi: 10.1099/00222615-8-3-413. [DOI] [PubMed] [Google Scholar]
  15. Ota F., Morita J., Yoshida N., Ashton F., Diena B. Studies on gonococcal infection. I. Electron microscopic studies on phagocytosis of Neisseria gonorrhoeae by macrophages. Jpn J Microbiol. 1975 Apr;19(2):149–155. doi: 10.1111/j.1348-0421.1975.tb00861.x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Ovcinnikov N. M., Delektorskij V. V. Electron microscope studies of gonococci in the urethral secretions of patients with gonorrhoea. Br J Vener Dis. 1971 Dec;47(6):419–439. doi: 10.1136/sti.47.6.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Punsalang A. P., Jr, Sawyer W. D. Role of pili in the virulence of Neisseria gonorrhoeae. Infect Immun. 1973 Aug;8(2):255–263. doi: 10.1128/iai.8.2.255-263.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith H. Microbial interference with host defence mechanisms. Monogr Allergy. 1975;9:13–38. [PubMed] [Google Scholar]
  21. Spink W. W., Keefer C. S. STUDIES OF GONOCOCCAL INFECTION. I. A STUDY OF THE MODE OF DESTRUCTION OF THE GONOCOCCUS IN VITRO. J Clin Invest. 1937 Mar;16(2):169–176. doi: 10.1172/JCI100844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stone J. L., Vernon S. K., Warren G. H. Cultivation and survival studies of Neisseria gonorrhoeae in a human diploid cell strain. Yale J Biol Med. 1974 Dec;47(4):291–296. [PMC free article] [PubMed] [Google Scholar]
  23. Swanson J., King G., Zeligs B. Studies on gonococcus infection. VIII. 125Iodine labeling of gonococci and studies on their in vitro interactions with eukaryotic cells. Infect Immun. 1975 Mar;11(3):453–459. doi: 10.1128/iai.11.3.453-459.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Swanson J., Sparks E., Young D., King G. Studies on Gonococcus infection. X. Pili and leukocyte association factor as mediators of interactions between gonococci and eukaryotic cells in vitro. Infect Immun. 1975 Jun;11(6):1352–1361. doi: 10.1128/iai.11.6.1352-1361.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Swanson J., Sparks E., Zeligs B., Siam M. A., Parrott C. Studies on gonococcus infection. V. Observations on in vitro interactions of gonococci and human neutrophils. Infect Immun. 1974 Sep;10(3):633–644. doi: 10.1128/iai.10.3.633-644.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Swanson J. Studies on gonococcus infection. IV. Pili: their role in attachment of gonococci to tissue culture cells. J Exp Med. 1973 Mar 1;137(3):571–589. doi: 10.1084/jem.137.3.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. THAYER J. D., PERRY M. I., FIELD F. W., GARSON W. Failure of penicillin, chloramphenicol, erythromycin, and novobiocin to kill phagocytized gonococci in tissue. Antibiot Annu. 1956:513–517. [PubMed] [Google Scholar]
  28. Thomas D. W., Hill J. C., Tyeryar F. J., Jr Interaction of gonococci with phagocytic leukocytes from men and mice. Infect Immun. 1973 Jul;8(1):98–104. doi: 10.1128/iai.8.1.98-104.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thongthai C., Sawyer W. D. Studies on the virulence of Neisseria gonorrhoeae. I. Relation of colonial morphology and resistance to phagocytosis by polymorphonuclear leukocytes. Infect Immun. 1973 Mar;7(3):373–379. doi: 10.1128/iai.7.3.373-379.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. 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]
  33. Watt P. J. The fate of gonococci in polymorphonuclear leucocytes. J Med Microbiol. 1970 Aug;3(3):501–509. doi: 10.1099/00222615-3-3-501. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES