Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1986 Dec;54(3):855–863. doi: 10.1128/iai.54.3.855-863.1986

Ultrastructural study of mode of entry of Chlamydia psittaci into L-929 cells.

R L Hodinka, P B Wyrick
PMCID: PMC260249  PMID: 2430888

Abstract

The entry of Chlamydia psittaci into L-929 cells was studied morphologically by transmission electron microscopy and quantitatively by a method that discriminates between attachment and uptake. Upon adsorption of 3H-labeled elementary bodies (EBs) to host cells at 4 degrees C, the EBs bound efficiently to the L-cell surface. Binding reached an equilibrium level of 55% in 3 h. Ultrastructural analysis revealed that EBs were bound preferentially to the tips and sides of microvilli at this temperature. The EBs were also observed in coated pits located at the bases of microvilli and along smooth surfaces of the host cell. No internalization was observed at 4 degrees C. When cells with prebound 3H-labeled EBs were warmed to 37 degrees C, the EBs rapidly became resistant to proteinase K removal (half time = 5 min), indicating ingested chlamydiae. At 37 degrees C, the EBs were internalized within tightly bound vesicles surrounded by an electron-dense coat of fibrillar material. EBs were also present in smooth-surfaced pits and vesicles of the host cell. Using alpha 2-macroglobulin coupled to colloidal gold (a known marker for receptor-mediated endocytosis), we observed that the entry of EBs into cells via coated pits was identical in appearance to the internalization of alpha 2-macroglobulin. Also, when the two ligands were mixed together, they could be seen within the same coated pits and were cointernalized within endocytic vesicles of the host cell. These results suggest that C. psittaci can enter nonprofessional phagocytic cells by a pathway which is similar to that of receptor-mediated endocytosis of many physiologically important macromolecules, bacterial toxins, and viruses.

Full text

PDF
855

Images in this article

857Image
on p.857
859Image
on p.859
860Image
on p.860
861Image
on p.861
861Image
on p.861
862Image
on p.862

Selected References

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

  1. Aggeler J., Werb Z. Initial events during phagocytosis by macrophages viewed from outside and inside the cell: membrane-particle interactions and clathrin. J Cell Biol. 1982 Sep;94(3):613–623. doi: 10.1083/jcb.94.3.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker Y. The chlamydia: molecular biology of procaryotic obligate parasites of eucaryocytes. Microbiol Rev. 1978 Jun;42(2):274–306. doi: 10.1128/mr.42.2.274-306.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brownridge E., Wyrick P. B. Interaction of Chlamydia psittaci reticulate bodies with mouse peritoneal macrophages. Infect Immun. 1979 Jun;24(3):697–700. doi: 10.1128/iai.24.3.697-700.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Byrne G. I., Moulder J. W. Parasite-specified phagocytosis of Chlamydia psittaci and Chlamydia trachomatis by L and HeLa cells. Infect Immun. 1978 Feb;19(2):598–606. doi: 10.1128/iai.19.2.598-606.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Darougar S., Cubitt S., Jones B. R. Effect of high-speed centrifugation on the sensitivity of irradiated McCoy cell culture for the isolation of Chlamydia. Br J Vener Dis. 1974 Aug;50(4):308–312. doi: 10.1136/sti.50.4.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickson R. B., Willingham M. C., Pastan I. alpha 2-macroglobulin adsorbed to colloidal gold: a new probe in the study of receptor-mediated endocytosis. J Cell Biol. 1981 Apr;89(1):29–34. doi: 10.1083/jcb.89.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doughri A. M., Storz J., Altera K. P. Mode of entry and release of chlamydiae in infections of intestinal epithelial cells. J Infect Dis. 1972 Dec;126(6):652–657. doi: 10.1093/infdis/126.6.652. [DOI] [PubMed] [Google Scholar]
  8. Eissenberg L. G., Wyrick P. B., Davis C. H., Rumpp J. W. Chlamydia psittaci elementary body envelopes: ingestion and inhibition of phagolysosome fusion. Infect Immun. 1983 May;40(2):741–751. doi: 10.1128/iai.40.2.741-751.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eissenberg L. G., Wyrick P. B. Inhibition of phagolysosome fusion is localized to Chlamydia psittaci-laden vacuoles. Infect Immun. 1981 May;32(2):889–896. doi: 10.1128/iai.32.2.889-896.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friis R. R. Interaction of L cells and Chlamydia psittaci: entry of the parasite and host responses to its development. J Bacteriol. 1972 May;110(2):706–721. doi: 10.1128/jb.110.2.706-721.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Geoghegan W. D., Ackerman G. A. Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem. 1977 Nov;25(11):1187–1200. doi: 10.1177/25.11.21217. [DOI] [PubMed] [Google Scholar]
  12. Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
  13. Gregory W. W., Byrne G. I., Gardner M., Moulder J. W. Cytochalasin B does not inhibit ingestion of Chlamydia psittaci by mouse fibroblasts (L cells) and mouse peritoneal macrophages. Infect Immun. 1979 Jul;25(1):463–466. doi: 10.1128/iai.25.1.463-466.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Helenius A., Kartenbeck J., Simons K., Fries E. On the entry of Semliki forest virus into BHK-21 cells. J Cell Biol. 1980 Feb;84(2):404–420. doi: 10.1083/jcb.84.2.404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Helenius A., Morein B., Fries E., Simons K., Robinson P., Schirrmacher V., Terhorst C., Strominger J. L. Human (HLA-A and HLA-B) and murine (H-2K and H-2D) histocompatibility antigens are cell surface receptors for Semliki Forest virus. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3846–3850. doi: 10.1073/pnas.75.8.3846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heuser J. Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol. 1980 Mar;84(3):560–583. doi: 10.1083/jcb.84.3.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuo C. C., Grayston T. Interaction of Chlamydia trachomatis organisms and HeLa 229 cells. Infect Immun. 1976 Apr;13(4):1103–1109. doi: 10.1128/iai.13.4.1103-1109.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kuo C. C., Wang S. P., Grayston J. T. Effect of polycations, polyanions and neuraminidase on the infectivity of trachoma-inclusin conjunctivitis and lymphogranuloma venereum organisms HeLa cells: sialic acid residues as possible receptors for trachoma-inclusion conjunction. Infect Immun. 1973 Jul;8(1):74–79. doi: 10.1128/iai.8.1.74-79.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Matlin K. S., Reggio H., Helenius A., Simons K. Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981 Dec;91(3 Pt 1):601–613. doi: 10.1083/jcb.91.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Middlebrook J. L., Dorland R. B. Bacterial toxins: cellular mechanisms of action. Microbiol Rev. 1984 Sep;48(3):199–221. doi: 10.1128/mr.48.3.199-221.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Morris R. E., Gerstein A. S., Bonventre P. F., Saelinger C. B. Receptor-mediated entry of diphtheria toxin into monkey kidney (Vero) cells: electron microscopic evaluation. Infect Immun. 1985 Dec;50(3):721–727. doi: 10.1128/iai.50.3.721-727.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salisbury J. L., Condeelis J. S., Satir P. Receptor-mediated endocytosis: machinery and regulation of the clathrin-coated vesicle pathway. Int Rev Exp Pathol. 1983;24:1–62. [PubMed] [Google Scholar]
  23. Schachter J., Caldwell H. D. Chlamydiae. Annu Rev Microbiol. 1980;34:285–309. doi: 10.1146/annurev.mi.34.100180.001441. [DOI] [PubMed] [Google Scholar]
  24. Silverstein S. C., Steinman R. M., Cohn Z. A. Endocytosis. Annu Rev Biochem. 1977;46:669–722. doi: 10.1146/annurev.bi.46.070177.003321. [DOI] [PubMed] [Google Scholar]
  25. Simons K., Garoff H., Helenius A. How an animal virus gets into and out of its host cell. Sci Am. 1982 Feb;246(2):58–66. doi: 10.1038/scientificamerican0282-58. [DOI] [PubMed] [Google Scholar]
  26. Storz J., Spears P. Chlamydiales: properties, cycle of development and effect on eukaryotic host cells. Curr Top Microbiol Immunol. 1977;76:167–214. doi: 10.1007/978-3-642-66653-7_5. [DOI] [PubMed] [Google Scholar]
  27. Söderlund G., Kihlström E. Effect of methylamine and monodansylcadaverine on the susceptibility of McCoy cells to Chlamydia trachomatis infection. Infect Immun. 1983 May;40(2):534–541. doi: 10.1128/iai.40.2.534-541.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Söderlund G., Kihlström E. Physicochemical surface properties of elementary bodies from different serotypes of chlamydia trachomatis and their interaction with mouse fibroblasts. Infect Immun. 1982 Jun;36(3):893–899. doi: 10.1128/iai.36.3.893-899.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. TAMURA A., HIGASHI N. PURIFICATION AND CHEMICAL COMPOSITION OF MENINGOPNEUMONITIS VIRUS. Virology. 1963 Aug;20:596–604. doi: 10.1016/0042-6822(63)90284-8. [DOI] [PubMed] [Google Scholar]
  30. Ward M. E. Chlamydial classification, development and structure. Br Med Bull. 1983 Apr;39(2):109–115. doi: 10.1093/oxfordjournals.bmb.a071800. [DOI] [PubMed] [Google Scholar]
  31. Ward M. E., Murray A. Control mechanisms governing the infectivity of Chlamydia trachomatis for HeLa cells: mechanisms of endocytosis. J Gen Microbiol. 1984 Jul;130(7):1765–1780. doi: 10.1099/00221287-130-7-1765. [DOI] [PubMed] [Google Scholar]
  32. Wyrick P. B., Brownridge E. A. Growth of Chlamydia psittaci in macrophages. Infect Immun. 1978 Mar;19(3):1054–1060. doi: 10.1128/iai.19.3.1054-1060.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wyrick P. B., Brownridge E. A., Ivins B. E. Interaction of Chlamydia psittaci with mouse peritoneal macrophages. Infect Immun. 1978 Mar;19(3):1061–1067. doi: 10.1128/iai.19.3.1061-1067.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zeichhardt H., Wetz K., Willingmann P., Habermehl K. O. Entry of poliovirus type 1 and Mouse Elberfeld (ME) virus into HEp-2 cells: receptor-mediated endocytosis and endosomal or lysosomal uncoating. J Gen Virol. 1985 Mar;66(Pt 3):483–492. doi: 10.1099/0022-1317-66-3-483. [DOI] [PubMed] [Google Scholar]

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

RESOURCES