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Chlamydia trachomatis in cell culture

II. Susceptibility of seven established mammalian cell types in vitro. Adaptation of trachoma organisms to McCoy and BHK-21 cells

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Trachoma organisms of serotype B were grown serially in irradiated cells (McCoy, BHK-21, Microbiological Associates, and BHK-21, Lister) and tested for infectivity in monolayers of five mammalian cell lines (BHK-21, CHO, HeLa S3, McCoy and OWMK) and two diploid strains (ST/BTL and WI-38). All cell types had low susceptibility to chlamydial infection but the number of inclusions increased when the inoculum was centrifuged onto the monolayers, or when the cells were irradiated. Infection was higher in non-irradiated CHO than in irradiated CHO in three out of a total of six experiments. Inclusion number was increased 300 times in HeLa S3 and up to three times in the other cell types after treatment with diethylaminoethyl-dextran (DEAE-D). Serial passage of Chlamydia trachomatis serotype B (strain Har-36) in CO60 McCoy and CO60 BHK-21 Lister resulted in partial adaptation of the strain to the host cell. The phenomenon of adaptation of serotype B to McCoy compensated for the lower susceptibility of this cell revealed when McCoy cells were inoculated with trachoma elementary bodies grown in BHK-21 Lister or in chick embryo yolk sac. Trachoma organisms of immunotypes A, B and C prepared in yolk sac produced more inclusion-forming units per ml in CO60 BHK-21 Lister than in CO60 McCoy.

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References

  1. Storz, J. 1971.Chlamydia and Chlamydis-Induced Diseases. Charles C. Tomas, Springfield, Ill., pp. 81–83.

    Google Scholar 

  2. Blyth, W. A., and J. Taverne. 1974. Cultivation of TRIC agents: a comparison between the use of BHK-21 and irradiated McCoy cells. J. Hyg. (Camb.) 72: 121–128.

    CAS  Google Scholar 

  3. Croy, T. R., C. C. Kuo, and S-P Wang. 1975. Comparative susceptibility of eleven mammalian cell lines to infection with trachoma organisms. J. Clin. Microbiol. 1: 434–439.

    PubMed  CAS  Google Scholar 

  4. Gordon, F. B., H. R. Dressler, A. L. Quan, W. T. and J. I. Thomas. 1972. Effect of ionizing irradiation on susceptibility of McCoy cell cultures to chlamydia trachomatis. Appl. Micribiol. 23: 123–129.

    CAS  Google Scholar 

  5. Reeve, P., and J. Taverne. 1967. Strain differences in the behavior of TRIC agents in cell culture. Am. J. Ophthalmol. 63: 141–147.

    Google Scholar 

  6. Jenkin, H. M. 1966. The continuous passage of agents of trachoma in cell culture. I. Characteristics of TW-3 and Bour strains of trachoma cultivated in serial passage in HeLa 229. J. Infect. Dis. 116: 390–399.

    PubMed  CAS  Google Scholar 

  7. Kuo, C. C., S-P Wang, and J. T. Grayston. 1972. Differentiation of TRIC and LGV organisms based on enhancement of infection by DEAE-dextran in cell culture. J. Infect. Dis. 125: 313–317.

    PubMed  CAS  Google Scholar 

  8. Kuo, C. C., S-P Wang, and J. T. Grayston. 1973. Effect on polycations, polyanions, and neur-aminidase on the infectivity of trachoma-inclusion conjunctivitis and lymphogranuloma venereum organisms in HeLa cells: sialic acid residues as possible receptors for trachoma-inclusion conjunctivitis. Infect. Immun. 8: 74–79.

    PubMed  CAS  Google Scholar 

  9. Kuo, C. C., and J. T. Grayston. 1976. Interaction of chlamydia trachomatis organisms in HeLa 229 cells. Infect. Immun. 13: 1103–1109.

    PubMed  CAS  Google Scholar 

  10. Darougar, S., J. R. Kinnison, and B. R. Jones. 1971. Simplified irradiated McCoy cell culture for isolation of chlamydiae. In: R. L. Nichols (Ed.),Trachoma and Related Disorders Caused by Chlamydial Agents. Excerpta Medica, Princeton, pp. 63–70.

    Google Scholar 

  11. Rota, T. R., and R. L. Nichols. 1971. Infection of cell cultures by trachoma agents: enhancement by DEAE-dextran. J. Infect. Dis. 124: 19–421.

    Google Scholar 

  12. Rota, T. R., and R. L. Nichols. 1973.Chlamydia trachomatis in cell culture. I. Comparisons of efficiencies of infection in several chemically defined media, at various pH and temperature values, and after exposure to DEAE-dextran. Appl. Microbiol. 26: 560–565.

    PubMed  CAS  Google Scholar 

  13. Wentworth, B. B., P. Bonin, K. K. Holmes, L. Gutman, P. Wiesner, and E. R. Alexander. 1973. Isolation of viruses, bacteria and other organisms from venereal disease clinic patients: methodology and problems associated with multiple isolations. Health Lab. Sci. 10: 75–81.

    PubMed  CAS  Google Scholar 

  14. Gordon, F. B., and A. L. Quan. 1965. Isolation of the trachoma agent in cell culture. Proc. Soc. Exp. Biol. Med. 118: 354–359.

    PubMed  CAS  Google Scholar 

  15. Yerganian, G., and K. S. Lavappa. 1971. Procedures for culturing diploid cells and preparations of meiotic chromosomes from dwarf species of hamsters. In: A. Hollaender (Ed.),Chemical Mutagens: Principles and Methods for Their Detection. Vol. 2. Plenum Press, New York, pp. 387–410.

    Google Scholar 

  16. Tjio, J. H., and T. T. Puck. 1958. Genetics of somatic mammalian cells. II. Chromosomal constitution of cells in tissue culture. J. Exp. Med. 108: 259–268.

    Article  PubMed  CAS  Google Scholar 

  17. Kraemer, P. M., D. F. Petersen, and M. A. Van Dilla. 1971. DNA constancy in heteroploidy and the stem-line theory of tumors. Science. 174: 714–717.

    Article  PubMed  CAS  Google Scholar 

  18. Petersen, D. F., E. C. Anderson, and R. A. Tobey, 1968. Chapter 15. In: D. M. Prescott (Ed.),Methods in Cell Physiology. Vol. III. Academic Press, New York, pp. 347–370.

    Google Scholar 

  19. Hayflick, L. 1965. The limitedin vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614–636.

    Article  PubMed  CAS  Google Scholar 

  20. Stoker, M., and I. Macpherson. 1964. Syrian hamster fibroblast cells, line BHK-21 and its derivatives. Nature 203: 1355–1357.

    Article  PubMed  CAS  Google Scholar 

  21. Capstick, P. B. 1963. Growth of baby hamster kidney cells in suspension. Proc. Royal Soc. Med. 56: 1062–1064.

    CAS  Google Scholar 

  22. Macpherson, I., and M. Stoker. 1966. United States Patent No. 3,228,840.

  23. Puck, T. T., Ph.I. Marcus, and S. Ciecura. 1956. Clonal growth of mammalian cellsin vitro. Growth characteristics of colonies from single HeLa cells with and without a “feeder layer”. J. Exp. Med. 103: 273–284.

    Article  PubMed  CAS  Google Scholar 

  24. Gordon, F. B., G. B. Magruder, A. L. Quan, and H. G. Arm. 1963. Cell cultures for detection of trachoma virus from experimental simian infections. Proc. Soc. Exp. Biol. Med. 112: 236–242.

    PubMed  CAS  Google Scholar 

  25. Daniel, M. D., D. Silva, and N. Ma. 1976. Establishment of owl monkey kidney-210 cell line for virological studies (abstr.). In Vitro 12: 290.

    Google Scholar 

  26. Ham, R. H. 1962. An improved nutrient solution for diploid Chine hamster and human cell lines. Exp. Cell Res. 29: 515–526.

    Article  Google Scholar 

  27. McComb, D. E., and C. Puzniak. 1974. Micro cell culture method for isolation ofchlamydia trachomatis. Appl. Microbiol. 28: 727–729.

    PubMed  CAS  Google Scholar 

  28. Vedros, N. A., and F. B. Gordon. 1963. The influence of glucose concentration on growth of P-T-L viruses in McCoy cells (abstr.). Bacteriol. Proc. p. 134.

  29. Kundsin, R. B. 1970. The role of mycoplasmas in human reproductive failure. Ann. N.Y. Acad. Sci. 174: 794–797.

    Article  PubMed  CAS  Google Scholar 

  30. Puck, T. T., and Ph.I. Marcus. 1956. Action of X-rays on mammalian cells. J. Exp. Med. 103: 653–666.

    Article  PubMed  CAS  Google Scholar 

  31. Harrison, M. J. 1970. Enhancing effect of DEAE-dextran on inclusion counts of an ovine chlamydia (bdsonia) in cell culture. Aust. J. Exp. Biol. Med. Sci. 42: 207–213.

    Google Scholar 

  32. Bell, S. D., Jr., D. E. McComb, R. L. Nichols, and M. Roca-Garcia. 1970. Studies of trachoma. VII. Isolation of a mixture of type I and type II trachoma strians from a child in Saudi Arabia. Amer. J. Trop. Med. Hyg. 19: 842–845.

    CAS  Google Scholar 

  33. Gear, J. H. S., F. B. Gordon, B. R. Jones, and S. D. Bell, Jr. 1963. Nomenclature of isolates from trachoma and inclusion blennorrhea. Nature 197; 26.

    Article  Google Scholar 

  34. Alexander, E. R., S-P Wang, and J. T. Grayston. 1967. Further classification of TRIC agents from ocular trachoma and other sources by the mouse toxicity prevention test. Am. J. Opthalmol. 63: 1469–1478.

    Google Scholar 

  35. Gordon, F. B., and A. L. Quan. 1965. Occurrence of glycogen in inclusions of the psittacosis-lymphogranuloma venereum-trachoma agents. J. Infect. Dis. 115: 186–196.

    PubMed  CAS  Google Scholar 

  36. Kajima, M., N. Sharon, and M. Pollard. 1967. Virus-like particles in cultures of McCoy cells. J. Ultrastruct. Res. 18: 661–666.

    Article  PubMed  CAS  Google Scholar 

  37. Wade, N. 1972. Division of Biological Standards: The boat that never rocked. Science 175: 1225–1230.

    Article  PubMed  Google Scholar 

  38. Wade, N. 1976. Vaccine cells found mostly contaminated. Science 194: 41.

    Article  PubMed  Google Scholar 

  39. Pan American Health Organization. 1971. Foot-and-mouth Disease and Zoonoses Control. III. Inter-American Meeting. Scientific Publication No. 218, Washington, D.C.

  40. Walters, R. A., and R. A. Tobey. 1970. Radio-sensitivity of mammalian cells. IV. Effects of x-radiation on the DNA synthetic period in synchronized cells. Biophys. J. 10: 556–562.

    Article  PubMed  CAS  Google Scholar 

  41. Bell, S. D., Jr., and C. E. O. Fraser. 1969. Experimental trachoma in owl monkeys. Am. J. Trop. Med. Hyg. 18: 568–572.

    PubMed  CAS  Google Scholar 

  42. McComb, D. E., J. H. Peters, C. E. O. Fraser, E. S. Murray, A. B. MacDonald, and R. L. Nichols. 1971. Resistance to trachoma infection in owl monkeys correlated with antibody status at the outset in an experiment to test the response to topical trachoma antigens. In: R. L. Nichols (Ed.),Trachoma and Related Disorder Caused by Chlamydial Agents. Excerpta Medica, Princeton, pp. 396–406.

    Google Scholar 

  43. Hahon, N., N. A. Booth, and H. Eckert. 1973. Cell attachment and penetration by influenza virus. Infect. Immun. 7: 341–351.

    PubMed  CAS  Google Scholar 

  44. Mitsui, Y., T. Kitamuro, and M. Fujimoto. 1967. Adaptation of TRIC agents to tissue culture and characteristics of tissue culture adapted variants. Am. J. Ophthalmol. 63: 165–179.

    Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, Harvard School of Public Health, 665 Huntington Avenue, 02115, Boston, Massachusetts

    Teresa Rondon Rota

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  1. Teresa Rondon Rota
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The paper is dedicated to the memory of Francis B. Gordon, who pioneered research methods for the cultivation of trachoma and inclusion conjunctivitis (TRIC) agents in cell culture. Dr. Gordon patiently studied tables and photographs which accompany this text when he visited our laboratory on the day prior to his sailing to England on the ill-fated voyage in which he and Mrs. Gordon perished (October 1973).

This research was supported by a grant from the National Eye Institute (EI-00812-08), and by the Arabian American Oil Company.

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Rota, T.R. Chlamydia trachomatis in cell culture. In Vitro 13, 280–292 (1977). https://doi.org/10.1007/BF02616172

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