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. 1988 Apr 1;167(4):1459–1471. doi: 10.1084/jem.167.4.1459

Role of hemolysin for the intracellular growth of Listeria monocytogenes

PMCID: PMC2188911  PMID: 2833557

Abstract

Listeria monocytogenes insertion mutants defective in hemolysin production were generated using the conjugative transposons Tn916 and Tn1545. All of the nonhemolytic mutants (hly-) lacked a secreted 58-kD polypeptide, presumedly hemolysin, and were avirulent in a mouse model. An intracellular multiplication assay was established in monolayers of mouse bone marrow-derived macrophages, the J774 macrophage-like cell line, the CL.7 embryonic mouse fibroblast cell line, and the Henle 407 human epithelial cell line. The hly+ strain grew intracellularly in all of the tissue culture cells with a doubling time of approximately 60 min. In contrast, the hly- mutants failed to grow in the murine-derived tissue culture cells, but retained the ability to grow in the human tissue culture cells examined. Hemolytic-positive revertants were selected after passage of the hly- mutants through monolayers of J774 cells. In each case, the hemolytic revertants possessed the 58-kD polypeptide, were capable of intracellular growth in tissue culture monolayers and were virulent for mice.

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Selected References

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  1. Adams D. O., Hamilton T. A. The cell biology of macrophage activation. Annu Rev Immunol. 1984;2:283–318. doi: 10.1146/annurev.iy.02.040184.001435. [DOI] [PubMed] [Google Scholar]
  2. Bancroft G. J., Bosma M. J., Bosma G. C., Unanue E. R. Regulation of macrophage Ia expression in mice with severe combined immunodeficiency: induction of Ia expression by a T cell-independent mechanism. J Immunol. 1986 Jul 1;137(1):4–9. [PubMed] [Google Scholar]
  3. Berche P., Gaillard J. L., Sansonetti P. J. Intracellular growth of Listeria monocytogenes as a prerequisite for in vivo induction of T cell-mediated immunity. J Immunol. 1987 Apr 1;138(7):2266–2271. [PubMed] [Google Scholar]
  4. Celada A., Gray P. W., Rinderknecht E., Schreiber R. D. Evidence for a gamma-interferon receptor that regulates macrophage tumoricidal activity. J Exp Med. 1984 Jul 1;160(1):55–74. doi: 10.1084/jem.160.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Centers for Disease Control (CDC) Listeriosis outbreak associated with Mexican-style cheese--California. MMWR Morb Mortal Wkly Rep. 1985 Jun 21;34(24):357–359. [PubMed] [Google Scholar]
  6. Cheers C., Sandrin M. S. Restriction in adoptive transfer of resistance to Listeria monocytogenes. II. Use of congenic and mutant mice show transfer to be H-2K restricted. Cell Immunol. 1983 Jun;78(2):199–205. doi: 10.1016/0008-8749(83)90274-5. [DOI] [PubMed] [Google Scholar]
  7. Czuprynski C. J., Henson P. M., Campbell P. A. Killing of Listeria monocytogenes by inflammatory neutrophils and mononuclear phagocytes from immune and nonimmune mice. J Leukoc Biol. 1984 Feb;35(2):193–208. doi: 10.1002/jlb.35.2.193. [DOI] [PubMed] [Google Scholar]
  8. Emmerling P., Finger H., Hof H. Cell-mediated resistance to infection with Listeria monocytogenes in nude mice. Infect Immun. 1977 Feb;15(2):382–385. doi: 10.1128/iai.15.2.382-385.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fleming D. W., Cochi S. L., MacDonald K. L., Brondum J., Hayes P. S., Plikaytis B. D., Holmes M. B., Audurier A., Broome C. V., Reingold A. L. Pasteurized milk as a vehicle of infection in an outbreak of listeriosis. N Engl J Med. 1985 Feb 14;312(7):404–407. doi: 10.1056/NEJM198502143120704. [DOI] [PubMed] [Google Scholar]
  10. Gaillard J. L., Berche P., Mounier J., Richard S., Sansonetti P. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun. 1987 Nov;55(11):2822–2829. doi: 10.1128/iai.55.11.2822-2829.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gaillard J. L., Berche P., Sansonetti P. Transposon mutagenesis as a tool to study the role of hemolysin in the virulence of Listeria monocytogenes. Infect Immun. 1986 Apr;52(1):50–55. doi: 10.1128/iai.52.1.50-55.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Geoffroy C., Gaillard J. L., Alouf J. E., Berche P. Purification, characterization, and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes. Infect Immun. 1987 Jul;55(7):1641–1646. doi: 10.1128/iai.55.7.1641-1646.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gray M. L., Killinger A. H. Listeria monocytogenes and listeric infections. Bacteriol Rev. 1966 Jun;30(2):309–382. doi: 10.1128/br.30.2.309-382.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hahn H., Kaufmann S. H. The role of cell-mediated immunity in bacterial infections. Rev Infect Dis. 1981 Nov-Dec;3(6):1221–1250. doi: 10.1093/clinids/3.6.1221. [DOI] [PubMed] [Google Scholar]
  15. Hale T. L., Bonventre P. F. Shigella infection of Henle intestinal epithelial cells: role of the bacterium. Infect Immun. 1979 Jun;24(3):879–886. doi: 10.1128/iai.24.3.879-886.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Havell E. A. Synthesis and secretion of interferon by murine fibroblasts in response to intracellular Listeria monocytogenes. Infect Immun. 1986 Dec;54(3):787–792. doi: 10.1128/iai.54.3.787-792.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hayes P. S., Feeley J. C., Graves L. M., Ajello G. W., Fleming D. W. Isolation of Listeria monocytogenes from raw milk. Appl Environ Microbiol. 1986 Feb;51(2):438–440. doi: 10.1128/aem.51.2.438-440.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Heesemann J., Laufs R. Double immunofluorescence microscopic technique for accurate differentiation of extracellularly and intracellularly located bacteria in cell culture. J Clin Microbiol. 1985 Aug;22(2):168–175. doi: 10.1128/jcm.22.2.168-175.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hof H. Virulence of different strains of Listeria monocytogenes serovar 1/2a. Med Microbiol Immunol. 1984;173(4):207–218. doi: 10.1007/BF02122112. [DOI] [PubMed] [Google Scholar]
  20. JENKINS E. M., NJOKU-OBI A. N., ADAMS E. W. PURIFICATION OF THE SOLUBLE HEMOLYSINS OF LISTERIA MONOCYTOGENES. J Bacteriol. 1964 Aug;88:418–424. doi: 10.1128/jb.88.2.418-424.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kathariou S., Metz P., Hof H., Goebel W. Tn916-induced mutations in the hemolysin determinant affecting virulence of Listeria monocytogenes. J Bacteriol. 1987 Mar;169(3):1291–1297. doi: 10.1128/jb.169.3.1291-1297.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kaufmann S. H., Hug E., De Libero G. Listeria monocytogenes-reactive T lymphocyte clones with cytolytic activity against infected target cells. J Exp Med. 1986 Jul 1;164(1):363–368. doi: 10.1084/jem.164.1.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaufmann S. H., Simon M. M., Hahn H. Specific Lyt 123 cells are involved in protection against Listeria monocytogenes and in delayed-type hypersensitivity to listerial antigens. J Exp Med. 1979 Oct 1;150(4):1033–1038. doi: 10.1084/jem.150.4.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kingdon G. C., Sword C. P. Biochemical and Immunological Effects of Listeria monocytogenes Hemolysin. Infect Immun. 1970 Apr;1(4):363–372. doi: 10.1128/iai.1.4.363-372.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kingdon G. C., Sword C. P. Effects of Listeria monocytogenes Hemolysin on Phagocytic Cells and Lysosomes. Infect Immun. 1970 Apr;1(4):356–362. doi: 10.1128/iai.1.4.356-362.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Lane F. C., Unanue E. R. Requirement of thymus (T) lymphocytes for resistance to listeriosis. J Exp Med. 1972 May 1;135(5):1104–1112. doi: 10.1084/jem.135.5.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lepay D. A., Steinman R. M., Nathan C. F., Murray H. W., Cohn Z. A. Liver macrophages in murine listeriosis. Cell-mediated immunity is correlated with an influx of macrophages capable of generating reactive oxygen intermediates. J Exp Med. 1985 Jun 1;161(6):1503–1512. doi: 10.1084/jem.161.6.1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. MACKANESS G. B. Cellular resistance to infection. J Exp Med. 1962 Sep 1;116:381–406. doi: 10.1084/jem.116.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. MacDonald T. T., Carter P. B. Cell-mediated immunity to intestinal infection. Infect Immun. 1980 May;28(2):516–523. doi: 10.1128/iai.28.2.516-523.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. NJOKU-OBI A. N., JENKINS E. M., NJOKU-OBI J. C., ADAMS J., COVINGTON V. PRODUCTION AND NATURE OF LISTERIA MONOCYTOGENES HEMOLYSINS. J Bacteriol. 1963 Jul;86:1–8. doi: 10.1128/jb.86.1.1-8.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. North R. J. Importance of thymus-derived lymphocytes in cell-mediated immunity to infection. Cell Immunol. 1973 Apr;7(1):166–176. doi: 10.1016/0008-8749(73)90193-7. [DOI] [PubMed] [Google Scholar]
  33. North R. J. Suppression of cell-mediated immunity to infection by an antimitotic drug. Further evidence that migrant macrophages express immunity. J Exp Med. 1970 Sep 1;132(3):535–545. doi: 10.1084/jem.132.3.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. North R. J. The relative importance of blood monocytes and fixed macrophages to the expression of cell-mediated immunity to infection. J Exp Med. 1970 Sep 1;132(3):521–534. doi: 10.1084/jem.132.3.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ralph P., Prichard J., Cohn M. Reticulum cell sarcoma: an effector cell in antibody-dependent cell-mediated immunity. J Immunol. 1975 Feb;114(2 Pt 2):898–905. [PubMed] [Google Scholar]
  36. Rácz P., Tenner K., Mérö E. Experimental Listeria enteritis. I. An electron microscopic study of the epithelial phase in experimental listeria infection. Lab Invest. 1972 Jun;26(6):694–700. [PubMed] [Google Scholar]
  37. Sword C. P. Mechanisms of pathogenesis in Listeria monocytogenes infection. I. Influence of iron. J Bacteriol. 1966 Sep;92(3):536–542. doi: 10.1128/jb.92.3.536-542.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weis J., Seeliger H. P. Incidence of Listeria monocytogenes in nature. Appl Microbiol. 1975 Jul;30(1):29–32. doi: 10.1128/am.30.1.29-32.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weiss A. A., Hewlett E. L., Myers G. A., Falkow S. Tn5-induced mutations affecting virulence factors of Bordetella pertussis. Infect Immun. 1983 Oct;42(1):33–41. doi: 10.1128/iai.42.1.33-41.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Welshimer H. J., Donker-Voet J. Listeria monocytogenes in nature. Appl Microbiol. 1971 Mar;21(3):516–519. doi: 10.1128/am.21.3.516-519.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zachar Z., Savage D. C. Microbial interference and colonization of the murine gastrointestinal tract by Listeria monocytogenes. Infect Immun. 1979 Jan;23(1):168–174. doi: 10.1128/iai.23.1.168-174.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Zinkernagel R. M., Althage A., Adler B., Blanden R. V., Davidson W. F., Kees U., Dunlop M. B., Shreffler D. C. H-2 restriction of cell-mediated immunity to an intracellular bacterium: effector T cells are specific for Listeria antigen in association with H-21 region-coded self-markers. J Exp Med. 1977 May 1;145(5):1353–1367. doi: 10.1084/jem.145.5.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. von Koenig C. H., Finger H., Hof H. Failure of killed Listeria monocytogenes vaccine to produce protective immunity. Nature. 1982 May 20;297(5863):233–234. doi: 10.1038/297233a0. [DOI] [PubMed] [Google Scholar]

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