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. 1976 Oct;4(4):360–371. doi: 10.1128/jcm.4.4.360-371.1976

Examination of various cell culture techniques for co-incubation of virulent Treponema pallidum (Nichols I strain) under anaerobic conditions.

P L Sandok, S T Knight, H M Jenkin
PMCID: PMC274473  PMID: 789395

Abstract

Treponema pallidum (Nichols virulent) was incubated with and without cells in cell culture medium reduced to -275 mV Ecal, pH 7.3, under deoxygenated conditions. Five to ten percent of the treponemes attached to cells and remained motile for at least 120 h in cell-treponeme systems of co-incubation. Virulent treponemes could be detected after 120 to 144 h in the supernatant fluids of cell-treponeme co-incubation cultures and in cell-free tubes containing medium harvested from aerobically cultivated mammalian cells. Medium supplemented with ox serum ultrafiltrate, pyruvate, and sodium thioglycolate and gas mixtures containing H2 and CO2 enhanced treponemal survival. Increases in treponemal numbers were observed using dark-field microscopy but were not substantiated using the rabbit lesion test. Continuous passage of the treponeme was not achieved in vitro.

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Selected References

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

  1. Anderson L. E., Hill E. G., Jenkin H. M., Greene A. E., Nichols W. W. Characterization of miniature pig kidney cells and their resistance to chlamydial infection. Am J Vet Res. 1976 Feb;37(2):183–186. [PubMed] [Google Scholar]
  2. BEUTLER E., DURON O., KELLY B. M. Improved method for the determination of blood glutathione. J Lab Clin Med. 1963 May;61:882–888. [PubMed] [Google Scholar]
  3. Baseman J. B., Hayes N. S. Protein synthesis by Treponema pallidum extracted from infected rabbit tissue. Infect Immun. 1974 Dec;10(6):1350–1355. doi: 10.1128/iai.10.6.1350-1355.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baseman J. B., Nichols J. C., Hayes N. C. Virulent Treponema pallidum: aerobe or anaerobe. Infect Immun. 1976 Mar;13(3):704–711. doi: 10.1128/iai.13.3.704-711.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cox C. D., Barber M. K. Oxygen uptake by Treponema pallidum. Infect Immun. 1974 Jul;10(1):123–127. doi: 10.1128/iai.10.1.123-127.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  7. Fitzgerald T. J., Miller J. N., Sykes J. A. Treponema pallidum (Nichols strain) in tissue cultures: cellular attachment, entry, and survival. Infect Immun. 1975 May;11(5):1133–1140. doi: 10.1128/iai.11.5.1133-1140.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Graves S. R., Sandok P. L., Jenkin H. M., Johnson R. C. Retention of motility and virulence of Treponema pallidum (Nichols strain) in vitro. Infect Immun. 1975 Nov;12(5):1116–1120. doi: 10.1128/iai.12.5.1116-1120.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grin E. I., Nadazdin M., Svob M. Effect of hyperbaric oxygen on experimental syphilis in the rabbit. Br J Vener Dis. 1973 Oct;49(5):405–412. doi: 10.1136/sti.49.5.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horvath I., Arko R. J., Bullard J. C. Effects of oxygen and nitrogen on the character of T. pallidum is subcutaneous chambers in mice. Br J Vener Dis. 1975 Oct;51(5):301–304. doi: 10.1136/sti.51.5.301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jones R. H., Finn M. A., Thomas J. J., Folger C. Growth and subculture of pathogenic T. pallidum (Nichols strain) in BHK-21 cultured tissue cells. Br J Vener Dis. 1976 Feb;52(1):18–23. doi: 10.1136/sti.52.1.18. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KIMM G. E., ALLEN R. H., MORTON H. J., MORGAN J. F. Enhancement of survival in vitro of Treponema pallidum by addition of glucose and magnesium. J Bacteriol. 1960 Nov;80:726–727. doi: 10.1128/jb.80.5.726-727.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. KIMM G. E., ALLEN R. H., MORTON H. J., MORGAN J. F. Studies on the in vitro survival of virulent Treponema pallidum. I. Methodology and basal synthetic medium. Am J Hyg. 1962 May;75:339–346. doi: 10.1093/oxfordjournals.aje.a120255. [DOI] [PubMed] [Google Scholar]
  14. Makino S., Jenkin H. M. Effect of fatty acids on growth of Japanese encephalitis virus cultivated in BHK-21 cells and phospholipid metabolism of the infected cells. J Virol. 1975 Mar;15(3):515–525. doi: 10.1128/jvi.15.3.515-525.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Metzger M., Smogór W. Study of the effect of pH and Eh values of the Nelson-Diesendruck medium on the survival of virulent Treponema pallidum. Arch Immunol Ther Exp (Warsz) 1966;14(4):445–453. [PubMed] [Google Scholar]
  16. NELSON R. A., Jr, DIESENDRUCK J. A., ZHEUTLIN H. E. C., STACK P. S., BARNETT M. Studies on treponemal immobilizing antibodies in syphilis. I. Techniques of measurement and factors influencing immobilization. J Immunol. 1951 Jun;66(6):667–685. [PubMed] [Google Scholar]
  17. Nichols J. C., Baseman J. B. Carbon sources utilized by virulent Treponema pallidum. Infect Immun. 1975 Nov;12(5):1044–1050. doi: 10.1128/iai.12.5.1044-1050.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ovcinnikov N. M., Delektorskij V. V. Electron microscopy of phagocytosis in syphilis and yaws. Br J Vener Dis. 1972 Aug;48(4):227–248. doi: 10.1136/sti.48.4.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. PORTNOY J., HARRIS A., OLANSKY S. Studies of the Treponema pallidum immobilization test. I. The effect of increased sodium thioglycollate and complement. Am J Syph Gonorrhea Vener Dis. 1953 Mar;37(2):101–105. [PubMed] [Google Scholar]
  20. RICE F. A. H., NELSON R. A., Jr The isolation from beef serum of a survival factor for Treponema pallidum. J Biol Chem. 1951 Jul;191(1):35–41. [PubMed] [Google Scholar]
  21. Rathlev T. Investigations on in vitro survival and virulence of T. pallidum under aerobiosis. Br J Vener Dis. 1975 Oct;51(5):296–300. doi: 10.1136/sti.51.5.296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Richter A., Sanford K. K., Evans V. J. Influence of oxygen and culture media on plating efficiency of some mammalian tissue cells. J Natl Cancer Inst. 1972 Dec;49(6):1705–1712. doi: 10.1093/jnci/49.6.1705. [DOI] [PubMed] [Google Scholar]
  23. Sandok P. L., Jenkin H. M., Graves S. R., Knight S. T. Retention of motility of Treponema pallidum (Nichols virulent strain) in an anaerobic cell culture system and in a cell-free system. J Clin Microbiol. 1976 Jan;3(1):72–74. doi: 10.1128/jcm.3.1.72-74.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sedlak J., Lindsay R. H. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 1968 Oct 24;25(1):192–205. doi: 10.1016/0003-2697(68)90092-4. [DOI] [PubMed] [Google Scholar]
  25. Shapiro H. M. Redox balance in the body: an approach to quantitation. J Surg Res. 1972 Sep;13(3):138–152. doi: 10.1016/0022-4804(72)90057-1. [DOI] [PubMed] [Google Scholar]
  26. Sykes J. A., Miller J. N. Intracellular location of Treponema pallidum (Nichols strain) in the rabbit testis. Infect Immun. 1971 Sep;4(3):307–314. doi: 10.1128/iai.4.3.307-314.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sykes J. A., Miller J. N., Kalan A. J. Treponema pallidum within cells of a primary chancre from a human female. Br J Vener Dis. 1974 Feb;50(1):40–44. doi: 10.1136/sti.50.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. WEBER M. M. Factors influencing the in vitro survival of Treponema pallidum. Am J Hyg. 1960 May;71:401–417. doi: 10.1093/oxfordjournals.aje.a120123. [DOI] [PubMed] [Google Scholar]
  29. Willcox R. R., Guthe T. Treponema pallidum. A bibliographical review of the morphology, culture and survival of T. pallidum and associated organisms. Bull World Health Organ. 1966;35:1–169. [PMC free article] [PubMed] [Google Scholar]

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