Aug 30, 1982 - Subsets in Listeria monocytogenes-Specific T-Cell Activation. STEFAN H. E. KAUFMANN,lt* MARKUS M. SIMON,2 AND HELMUT HAHN1.
Vol. 38, No. 3
INFECTION AND IMMUNITY, Dec. 1982, p. 907-913
0019-9567/82/120907-07$02.00/0 Copyright © 1982, American Society for Microbiology
Regulatory Interactions Between Macrophages and T-Cell Subsets in Listeria monocytogenes-Specific T-Cell Activation STEFAN H. E. KAUFMANN,lt* MARKUS M. SIMON,2 AND HELMUT HAHN1 Institut fur Medizinische Mikrobiologie der Freien Universitat Berlin, D-1000 Berlin 45,1 and Max-PlanckInstitut fur Immunbiologie, D-7800 Freiburg,2 Federal Republic of Germany Received 19 May 1982/Accepted 30 August 1982
Peritoneal exudate T lymphocytes from Listeria monocytogenes-immune mice in the presence of the homologous antigen (heat-killed L. monocytogenes) and normal macrophages showed L. monocytogenes-specific proliferative responses. Proliferation was inhibited by macrophages from L. monocytogenes- or Corynebacterium parvum-pretreated mice as well as by exogenous prostaglandin E2.
Macrophage-dependent inhibition of T-cell proliferation-at least in part-could be reversed by addition of indomethacin. When selected L. monocytogenesimmune Lyt T-cell subsets were cultured in the presence of inhibitory macrophages, pretreatment with anti-Lyt 1 antiserum plus complement completely abrogated proliferation and pretreatment with anti-Lyt 2 and anti-Lyt 3 antisera plus complement markedly reduced proliferation. However, a mixture (1:1) of the two preselected Lyt T-cell subsets resulted in complete reconstitution of proliferative responses. In contrast, when L. monocytogenes-immune peritoneal exudate T lymphocytes were treated with anti-Lyt antisera plus complement after culture, only treatment with anti-Lyt 1 antiserum plus complement affected proliferation, suggesting regulatory interactions between Lyt 1+23- and Lyt 1-23+ T cells during in vitro culture which result in proliferation within the Lyt 1+23- T-cell subset. After rigorous depletion of residual macrophages and in the presence of indomethacin, pretreatment with anti-Lyt 1 antiserum plus complement, but not with anti-Lyt 2 and 3 antisera plus complement, eliminated proliferation. The data presented indicate that interactions between macrophages and Lyt T-cell subsets regulate L. monocytogenes-specific T-cell activation.
Specific protection against facultative intracellular bacteria and delayed-type hypersensitivity (DTH) to their products are mediated by specific T lymphocytes (reviewed in reference 6). In mice, T cells of the Lyt 1+23+ subset are crucially involved in both protection and DTH (11). Also, protection and DTH are restricted by the H-21 locus of the major histocompatibility complex (9, 19). Recently, it was shown that in vitro interactions between normal macrophages and Listeria monocytogenes-immune T cells result in secretion of interleukins 1 and 2 (3, 10, 12; S. H. E. Kaufman, H. Hahn, and M. M. Simon, Scand. J. Immunol., in press) and that secretion of the two interleukins is induced by Lyt 1+23T cells (10, 12; Kaufman et al., in press). These data indicated collaborative interactions between Lyt 1+23- and Lyt 1+23+ T cells in the immune response to L. monocytogenes. The present report shows that macrophages, though essential for proliferative responses of L. t Present address: Max-Planck-Institut flir Immunbiologie,
StUbeweg 51 D-7800 Freiburg, Federal Republic of Gennany.
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monocytogenes-immune Lyt 1+23- T cells at high concentrations, are capable of inhibiting Tcell proliferation. Furthermore, evidence is presented that inhibitory macrophages are under the negative control of Lyt 1-23+ T cells. MATERIAULS AND METHODS Mice. (C57BL/6 x DBA/2)F1 (B6D2F1) mice were bred in our own facilities, and C57BL/6 mice were obtained from the Zentralinstitut ftir Versuchstierzucht, Hannover, Federal Republic of Germany. Mice were sex-matched in a given experiment and used at 8 to 12 weeks of age.
Bacteria and bacterial antigens. L. monocytogenes (strain EGD) was kept virulent by continuous mouse passage. Heat-killed organisms were used as listerial antigens (Kaufmann et al., in press). Heat-killed bacteria were stored at -70°C at concentrations of 5 x 108 organisms per ml. L. monocytogenes-immune T ceUs. Mice were immunized with approximately 5 x 103 live L. monocytogenes cells, and 7 days later peritoneal exudates were induced with 1.5 ml of 10%o proteose-peptone. After another 3 days, cells were harvested as described previously (Kaufmann et al., in press). Peritoneal exudate T lymphocytes (PETLs) were enriched by
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PETLs (x104) FI[G. 1. Dose dependence of L. monocytogenesspec-ific T-cell proliferation. Graded numbers of L. mon ocytogenes-immune PETLs were cocultured with 2 x 103 normal macrophages and 1 x 107 heat-killed L. montocytogenes cells in a total volume of 0.2 ml for 3 dayss. Data represent means of three determinations (staridard deviation,
