Abstract
The cell-mediated immune response to Listeria monocytogenes has been well characterized in the mouse. Listeriolysin O (LLO) is a major antigen in murine T-cell recognition of L. monocytogenes. In this study, we show that LLO is also recognized by human TcR alpha beta T cells and TcR gamma delta T cells. Human peripheral blood mononuclear cells (PBMC) cultured in vitro with live listeriae and then expanded with interleukin 2 were shown to respond to purified LLO. The generation of LLO-responsive T cells was dependent on the use of live bacteria during the initial in vitro challenge. LLO-induced proliferation of T cells expanded by exposure of PBMC to live listeriae was major histocompatibility complex restricted. PBMC cultured with formalin-fixed listeriae and subsequently expanded by interleukin 2 gave high proliferative responses to fixed bacteria but failed to respond to LLO. PBMC stimulated in vitro with fixed listeriae contained predominantly TcR alpha beta + T cells. In contrast, PBMC obtained from 85% of the donors studied generated high numbers of TcR gamma delta + T cells following in vitro culture with live listeriae. Using a panel of synthetic amphipathic LLO peptides, we found that LLO-specific T cells from different individuals recognized both common and unique peptides. LLO 470-508 was recognized by three of five individuals, while LLO 203-226 and LLO 107-126 were recognized by two of six individuals. A TcR gamma delta + T-cell line was established from PBMC stimulated with live listeriae and was shown to recognize LLO 470-508. Proliferative responses could be induced in this cell line by peptide-pulsed autologous PBMC but not by peptide-pulsed allogeneic PBMC. Our results establish the importance of LLO in human T-cell recognition of listeriae and show that both TcR alpha beta + T cells and TcR gamma delta + T cells recognize this antigen. Finally, since LLO 470-508 has a high degree of homology with other gram-positive bacterial toxins, the recognition of this peptide by TcR gamma delta + T cells suggests that an important role of these T cells in host defense is the recognition of bacterium-derived toxins.
Full Text
The Full Text of this article is available as a PDF (326.1 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Abo T., Sugawara S., Seki S., Fujii M., Rikiishi H., Takeda K., Kumagai K. Induction of human TCR gamma delta + and TCR gamma delta-CD2+CD3- double negative lymphocytes by bacterial stimulation. Int Immunol. 1990;2(8):775–785. doi: 10.1093/intimm/2.8.775. [DOI] [PubMed] [Google Scholar]
- Beattie I. A., Swaminathan B., Ziegler H. K. Cloning and characterization of T-cell-reactive protein antigens from Listeria monocytogenes. Infect Immun. 1990 Sep;58(9):2792–2803. doi: 10.1128/iai.58.9.2792-2803.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berche P., Gaillard J. L., Geoffroy C., Alouf J. E. T cell recognition of listeriolysin O is induced during infection with Listeria monocytogenes. J Immunol. 1987 Dec 1;139(11):3813–3821. [PubMed] [Google Scholar]
- 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]
- Bishop D. K., Hinrichs D. J. Adoptive transfer of immunity to Listeria monocytogenes. The influence of in vitro stimulation on lymphocyte subset requirements. J Immunol. 1987 Sep 15;139(6):2005–2009. [PubMed] [Google Scholar]
- Blanden R. V., Langman R. E. Cell-mediated immunity to bacterial infection in the mouse. Thymus-derived cells as effectors of acquired resistance to Listeria monocytogenes. Scand J Immunol. 1972;1(4):379–391. doi: 10.1111/j.1365-3083.1972.tb03304.x. [DOI] [PubMed] [Google Scholar]
- Bouwer H. G., Gibbins B. L., Jones S., Hinrichs D. J. Antilisterial immunity includes specificity to listeriolysin O (LLO) and non-LLO-derived determinants. Infect Immun. 1994 Mar;62(3):1039–1045. doi: 10.1128/iai.62.3.1039-1045.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bouwer H. G., Nelson C. S., Gibbins B. L., Portnoy D. A., Hinrichs D. J. Listeriolysin O is a target of the immune response to Listeria monocytogenes. J Exp Med. 1992 Jun 1;175(6):1467–1471. doi: 10.1084/jem.175.6.1467. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Camilli A., Paynton C. R., Portnoy D. A. Intracellular methicillin selection of Listeria monocytogenes mutants unable to replicate in a macrophage cell line. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5522–5526. doi: 10.1073/pnas.86.14.5522. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cluff C. W., Garcia M., Ziegler H. K. Intracellular hemolysin-producing Listeria monocytogenes strains inhibit macrophage-mediated antigen processing. Infect Immun. 1990 Nov;58(11):3601–3612. doi: 10.1128/iai.58.11.3601-3612.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cluff C. W., Ziegler H. K. Inhibition of macrophage-mediated antigen presentation by hemolysin-producing Listeria monocytogenes. J Immunol. 1987 Dec 1;139(11):3808–3812. [PubMed] [Google Scholar]
- Cossart P., Mengaud J. Listeria monocytogenes. A model system for the molecular study of intracellular parasitism. Mol Biol Med. 1989 Oct;6(5):463–474. [PubMed] [Google Scholar]
- Cossart P., Vicente M. F., Mengaud J., Baquero F., Perez-Diaz J. C., Berche P. Listeriolysin O is essential for virulence of Listeria monocytogenes: direct evidence obtained by gene complementation. Infect Immun. 1989 Nov;57(11):3629–3636. doi: 10.1128/iai.57.11.3629-3636.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Emoto M., Naito T., Nakamura R., Yoshikai Y. Different appearance of gamma delta T cells during salmonellosis between Ityr and Itys mice. J Immunol. 1993 Apr 15;150(8 Pt 1):3411–3420. [PubMed] [Google Scholar]
- 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]
- 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]
- Guo Y., Niesel D. W., Ziegler H. K., Klimpel G. R. Listeria monocytogenes activation of human peripheral blood lymphocytes: induction of non-major histocompatibility complex-restricted cytotoxic activity and cytokine production. Infect Immun. 1992 May;60(5):1813–1819. doi: 10.1128/iai.60.5.1813-1819.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haas W., Pereira P., Tonegawa S. Gamma/delta cells. Annu Rev Immunol. 1993;11:637–685. doi: 10.1146/annurev.iy.11.040193.003225. [DOI] [PubMed] [Google Scholar]
- Harty J. T., Bevan M. J. CD8+ T cells specific for a single nonamer epitope of Listeria monocytogenes are protective in vivo. J Exp Med. 1992 Jun 1;175(6):1531–1538. doi: 10.1084/jem.175.6.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hasegawa T., Tanaka T., Yoshikai Y. The appearance and role of gamma delta T cells in the peritoneal cavity and liver during primary infection with Listeria monocytogenes in rats. Int Immunol. 1992 Oct;4(10):1129–1136. doi: 10.1093/intimm/4.10.1129. [DOI] [PubMed] [Google Scholar]
- Havlir D. V., Ellner J. J., Chervenak K. A., Boom W. H. Selective expansion of human gamma delta T cells by monocytes infected with live Mycobacterium tuberculosis. J Clin Invest. 1991 Feb;87(2):729–733. doi: 10.1172/JCI115053. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hiromatsu K., Yoshikai Y., Matsuzaki G., Ohga S., Muramori K., Matsumoto K., Bluestone J. A., Nomoto K. A protective role of gamma/delta T cells in primary infection with Listeria monocytogenes in mice. J Exp Med. 1992 Jan 1;175(1):49–56. doi: 10.1084/jem.175.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holoshitz J., Koning F., Coligan J. E., De Bruyn J., Strober S. Isolation of CD4- CD8- mycobacteria-reactive T lymphocyte clones from rheumatoid arthritis synovial fluid. Nature. 1989 May 18;339(6221):226–229. doi: 10.1038/339226a0. [DOI] [PubMed] [Google Scholar]
- Holoshitz J., Vila L. M., Keroack B. J., McKinley D. R., Bayne N. K. Dual antigenic recognition by cloned human gamma delta T cells. J Clin Invest. 1992 Jan;89(1):308–314. doi: 10.1172/JCI115577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hsieh C. S., Macatonia S. E., Tripp C. S., Wolf S. F., O'Garra A., Murphy K. M. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science. 1993 Apr 23;260(5107):547–549. doi: 10.1126/science.8097338. [DOI] [PubMed] [Google Scholar]
- Inoue T., Yoshikai Y., Matsuzaki G., Nomoto K. Early appearing gamma/delta-bearing T cells during infection with Calmétte Guérin bacillus. J Immunol. 1991 Apr 15;146(8):2754–2762. [PubMed] [Google Scholar]
- Kabelitz D., Bender A., Schondelmaier S., Schoel B., Kaufmann S. H. A large fraction of human peripheral blood gamma/delta + T cells is activated by Mycobacterium tuberculosis but not by its 65-kD heat shock protein. J Exp Med. 1990 Mar 1;171(3):667–679. doi: 10.1084/jem.171.3.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaufmann S. H., Hahn H. Biological functions of t cell lines with specificity for the intracellular bacterium Listeria monocytogenes in vitro and in vivo. J Exp Med. 1982 Jun 1;155(6):1754–1765. doi: 10.1084/jem.155.6.1754. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Klimpel G. R., Niesel D. W., Asuncion M., Klimpel K. D. Natural killer cell activation and interferon production by peripheral blood lymphocytes after exposure to bacteria. Infect Immun. 1988 Jun;56(6):1436–1441. doi: 10.1128/iai.56.6.1436-1441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kozbor D., Trinchieri G., Monos D. S., Isobe M., Russo G., Haney J. A., Zmijewski C., Croce C. M. Human TCR-gamma+/delta+, CD8+ T lymphocytes recognize tetanus toxoid in an MHC-restricted fashion. J Exp Med. 1989 May 1;169(5):1847–1851. doi: 10.1084/jem.169.5.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kvalheim G., Sørensen O., Fodstad O., Funderud S., Kiesel S., Dörken B., Nustad K., Jakobsen E., Ugelstad J., Pihl A. Immunomagnetic removal of B-lymphoma cells from human bone marrow: a procedure for clinical use. Bone Marrow Transplant. 1988 Jan;3(1):31–41. [PubMed] [Google Scholar]
- Magee D. M., Wing E. J. Cloned L3T4+ T lymphocytes protect mice against Listeria monocytogenes by secreting IFN-gamma. J Immunol. 1988 Nov 1;141(9):3203–3207. [PubMed] [Google Scholar]
- Matar G. M., Bibb W. F., Helsel L., Dewitt W., Swaminathan B. Immunoaffinity purification, stabilization and comparative characterization of listeriolysin O from Listeria monocytogenes serotypes 1/2a and 4b. Res Microbiol. 1992 Jun;143(5):489–498. doi: 10.1016/0923-2508(92)90095-6. [DOI] [PubMed] [Google Scholar]
- Modlin R. L., Pirmez C., Hofman F. M., Torigian V., Uyemura K., Rea T. H., Bloom B. R., Brenner M. B. Lymphocytes bearing antigen-specific gamma delta T-cell receptors accumulate in human infectious disease lesions. Nature. 1989 Jun 15;339(6225):544–548. doi: 10.1038/339544a0. [DOI] [PubMed] [Google Scholar]
- Mombaerts P., Arnoldi J., Russ F., Tonegawa S., Kaufmann S. H. Different roles of alpha beta and gamma delta T cells in immunity against an intracellular bacterial pathogen. Nature. 1993 Sep 2;365(6441):53–56. doi: 10.1038/365053a0. [DOI] [PubMed] [Google Scholar]
- Moriwaki S., Korn B. S., Ichikawa Y., van Kaer L., Tonegawa S. Amino acid substitutions in the floor of the putative antigen-binding site of H-2T22 affect recognition by a gamma delta T-cell receptor. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11396–11400. doi: 10.1073/pnas.90.23.11396. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Munk M. E., Gatrill A. J., Kaufmann S. H. Target cell lysis and IL-2 secretion by gamma/delta T lymphocytes after activation with bacteria. J Immunol. 1990 Oct 15;145(8):2434–2439. [PubMed] [Google Scholar]
- Munk M. E., Kaufmann S. H. Listeria monocytogenes reactive T lymphocytes in healthy individuals. Microb Pathog. 1988 Jul;5(1):49–54. doi: 10.1016/0882-4010(88)90080-0. [DOI] [PubMed] [Google Scholar]
- North R. J. Cellular mediators of anti-Listeria immunity as an enlarged population of short lived, replicating T cells. Kinetics of their production. J Exp Med. 1973 Aug 1;138(2):342–355. doi: 10.1084/jem.138.2.342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohga S., Yoshikai Y., Takeda Y., Hiromatsu K., Nomoto K. Sequential appearance of gamma/delta- and alpha/beta-bearing T cells in the peritoneal cavity during an i.p. infection with Listeria monocytogenes. Eur J Immunol. 1990 Mar;20(3):533–538. doi: 10.1002/eji.1830200311. [DOI] [PubMed] [Google Scholar]
- Pamer E. G., Harty J. T., Bevan M. J. Precise prediction of a dominant class I MHC-restricted epitope of Listeria monocytogenes. Nature. 1991 Oct 31;353(6347):852–855. doi: 10.1038/353852a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Portnoy D. A., Jacks P. S., Hinrichs D. J. Role of hemolysin for the intracellular growth of Listeria monocytogenes. J Exp Med. 1988 Apr 1;167(4):1459–1471. doi: 10.1084/jem.167.4.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raulet D. H. The structure, function, and molecular genetics of the gamma/delta T cell receptor. Annu Rev Immunol. 1989;7:175–207. doi: 10.1146/annurev.iy.07.040189.001135. [DOI] [PubMed] [Google Scholar]
- Rosat J. P., MacDonald H. R., Louis J. A. A role for gamma delta + T cells during experimental infection of mice with Leishmania major. J Immunol. 1993 Jan 15;150(2):550–555. [PubMed] [Google Scholar]
- Safley S. A., Cluff C. W., Marshall N. E., Ziegler H. K. Role of listeriolysin-O (LLO) in the T lymphocyte response to infection with Listeria monocytogenes. Identification of T cell epitopes of LLO. J Immunol. 1991 May 15;146(10):3604–3616. [PubMed] [Google Scholar]
- Sasaki T., Mieno M., Udono H., Yamaguchi K., Usui T., Hara K., Shiku H., Nakayama E. Roles of CD4+ and CD8+ cells, and the effect of administration of recombinant murine interferon gamma in listerial infection. J Exp Med. 1990 Apr 1;171(4):1141–1154. doi: 10.1084/jem.171.4.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skalka B., Smola J., Elischerová K. Routine test for in vitro differentiation of pathogenic and apathogenic Listeria monocytogenes strains. J Clin Microbiol. 1982 Mar;15(3):503–507. doi: 10.1128/jcm.15.3.503-507.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skeen M. J., Ziegler H. K. Induction of murine peritoneal gamma/delta T cells and their role in resistance to bacterial infection. J Exp Med. 1993 Sep 1;178(3):971–984. doi: 10.1084/jem.178.3.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uyemura K., Klotz J., Pirmez C., Ohmen J., Wang X. H., Ho C., Hoffman W. L., Modlin R. L. Microanatomic clonality of gamma delta T cells in human leishmaniasis lesions. J Immunol. 1992 Feb 15;148(4):1205–1211. [PubMed] [Google Scholar]
- Vidović D., Roglić M., McKune K., Guerder S., MacKay C., Dembić Z. Qa-1 restricted recognition of foreign antigen by a gamma delta T-cell hybridoma. Nature. 1989 Aug 24;340(6235):646–650. doi: 10.1038/340646a0. [DOI] [PubMed] [Google Scholar]
- Yamamoto S., Russ F., Teixeira H. C., Conradt P., Kaufmann S. H. Listeria monocytogenes-induced gamma interferon secretion by intestinal intraepithelial gamma/delta T lymphocytes. Infect Immun. 1993 May;61(5):2154–2161. doi: 10.1128/iai.61.5.2154-2161.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]