Skip to main content
SpringerLink
Log in

Enhanced in vitro cytotoxic anti H-2 responses in the presence of Ia antigens

  • Published:
Immunogenetics Aims and scope Submit manuscript

Abstract

Because surface Ig and Ia antigens cap independently, A.TH anti-A.TL serum combined with the indirect immunfluorescence technique could be used to test defined murine cell populations ofH-2 k haplotype for the presence of Ia antigens. Mitogen induced T- and B-cell derived blast cells, purified by velocity sedimentation at 1g, were tested for the expression of Iak antigens and then used both as stimulator cells and as target cells, in primary and secondary in vitro cytotoxic allograft responses. Fibroblasts, cortisone-resistant thymocytes, and nylon column purified splenic T cells were also included in these tests. Ia antigens were detected on 100% of LPS-induced blast cells, on 20%–30% of ConA-induced blast cells (100%Θ Thy-1 or antigen positive), but only to 5%–10% on PHA-blasts (100% Thy-1 antigen positive). Fibroblasts and nylon column purified splenic T cells were essentially Ia negative. Ia-positive allogeneic stimulator cells induced a far stronger in vitro cytotoxic T-cell response compared to Ia-negative stimulator cells; that is, there was a positive correlation between the expression of Ia antigens on the stimulator cells and the magnitude of cytotoxicity induced. This correlation was restricted to primary allograft responses. Ia antigens could not be detected as a target for killing in the cytotoxic effector phase, using both different target cells as well as the approach of “PHA dependent lysis” for detecting cytotoxic T lymphocytes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

B cell:

bone-marrow derived lymphocyte

CML:

cell-mediated lymphocytotoxicity

Con A:

Concanavalin A

CTL:

cytotoxic T lymphocytes

FITC:

fluorescein isothiocyanate conjugated

GαRIg:

goat anti rabbit immunoglobulin

LAD:

lymphocyte activating determinants

LPS:

lipopolysaccharide

ME:

2-mercapthoethanol

MEM:

minimal essential medium

MLC:

mixed lymphocyte culture

MHC:

major histocompatibility complex

PHA:

phytohemagglutinin

RαMIg:

rabbit anti mouse immunoglobulin

s:

sedimentation rate

T cell:

thymus-derived lymphocyte

References

  • Bach, F.H., Widmer, M.B., Bach, M.L., and Klein, J.: Serologically defined and lymphocyte defined components of the major histocompatibility complex in the mouse.J. Exp. Med. 136:1430–1444, 1972

    Google Scholar 

  • Cerottini, J.C. and Brunner, K.T.: Cell-mediated cytotoxicity, allograft rejection and tumor immunity.Adv. Immunol. 18:67–132, 1974

    Google Scholar 

  • Cohen, L. and Howe, M.L.: Synergism between subpopulations of thymus derived cells mediating the proliferative and effector phase of the mixed lymphocyte reaction.Proc. Natl. Acad. Sci. USA 70:2707–2710, 1973

    Google Scholar 

  • Eijsvoogel, V., du Bois, M., Meinesz, A., Bierhorst, Eijlander, A., Zeylemaker, W., and Schelleskens, P.: The specificity and activation mechanism of cell-mediated lympholysis (CML) in man.Transplant Proc. 5:1675–1678, 1973

    Google Scholar 

  • Forman, J. and Möller, G.: Generation of cytotoxic lymphocytes in mixed lymphocyte reactions. I. Specificity of the effector cells.J. Exp. Med. 138:672–685, 1973

    Google Scholar 

  • Götze, D., Reisfeld, A., and Klein, J.: Serological evidence for antigens controlled by theIr-region in mice.J. Exp. Med. 138:1003–1008, 1973

    Google Scholar 

  • Hämmerling, G.J., Deak, B.D., Mauve, D., Hämmerling, U., and McDevitt, H.O.: B-lymphocyte alloantigens controlled by theI region of the major histocompatibility complex in mice.Immunogenetics 1:68–81, 1974

    Google Scholar 

  • Häyry, P. and Andersson, L.C.: T cell synergy in mixed lymphocyte culture induced cytolysis.Eur. J. Immunol. 4:145–146, 1974

    Google Scholar 

  • Häyry, P., Andersson, L.C., Nordling, S., and Virolainen, M.: Allograft response in vitro.Transplant. Rev. 12:91–140, 1972

    Google Scholar 

  • Hauptfeld, V., Klein, D., and Klein, J.: Serologic identification of anIr-region antigen.Science 181: 167–169, 1973

    Google Scholar 

  • Julius, M.H., Simpson, E., and Herzenberg, L.A.: A rapid method for the isolation of functional thymus-derived murine lymphocytes.Eur. J. Immunol. 3: 645–649, 1973

    Google Scholar 

  • Katz, O.H., Hamaoka, T., Dorf, E.M., and Benacerraf, B.: Cell interactions between histoincompatible T and B lymphocytes.Proc. Natl. Acad. Sci. USA 70:2624–2628, 1973

    Google Scholar 

  • Klein, J. and Park, J.M.: Graft-versus-host reaction across different regions of theH-2 complex of the mouse.J. Exp. Med. 137:1213–1225, 1973

    Google Scholar 

  • Klein, J., Hauptfeld, V., and Hauptfeld, M.: Involvement of H-2 regions in immune reactions. In L. Brent and J. Holborow (eds.):Progress in Immunology II, Vol 3; pp. 197–206, North-Holland Publ. Co. Amsterdam 1974.

    Google Scholar 

  • Klein, J., Hauptfeld, M., and Hauptfeld, V.: Serological distinction of mutants B6.C-H(zl) and B6.M505. from strain C57BL/6.J. exp. Med. 140:1127–1132, 1974

    Google Scholar 

  • Miller, R.G. and Phillips, R.A.: Separation of cells by velocity sedimentation.J. Cell Physiol. 73:191–195, 1969

    Google Scholar 

  • Möller, G.: Demonstration of mouse isoantigens at the cellular level by the fluorescent antibody technique.J. Exp. Med. 114:415–432, 1961

    Google Scholar 

  • Nabholz, M., Vives, J., Young, H.M., Miggiano, M.V., Rijnbeck, A., and Shreffler, D.C.: Cell-mediated cell lysis in vitro: Genetic control of killer cell production and target specificities in the mouse.Eur. J. Immunol. 4: 378–387, 1974

    Google Scholar 

  • Röllinghoff, M. and Wagner, H.: In vitro induction of tumor specific immunity: requirements for T lymphocytes and tumor growth inhibition in vivo.Eur. J. Immunol. 3:472–476, 1973

    Google Scholar 

  • Röllinghoff, M. and Wagner, H.: Secondary allograft responses in vitro: I. The antigenic requirements. Submitted for publication

  • Schendel, D.J. and Bach, F.H.: Genetic control of cell-mediated lympholysis in mouse.J. Exp. Med. 140: 1534–1546, 1974

    Google Scholar 

  • Shreffler, D.C., David, C.S., Götze, D., Klein, J., McDevitt, H.O., and Sachs, D.: Genetic nomenclature for new lymphocyte antigens controlled by the I region of theH-2 complex.Immunogenetics 1: 189–194, 1974

    Google Scholar 

  • Shreffler, D.C. and David, C.S.: TheH-2 major histocompatibility complex and theI immune response region: Genetic variation, function and organisation.Adv. Immunol. 20: 125–195, 1975

    Google Scholar 

  • Sprent, J. and Miller, J.F.A.P.: Interaction of thymus lymphocytes with histoincompatible cells: I. Quantitation of the proliferative response of thymus cells.Cell. Immunol. 3:361–384, 1972

    Google Scholar 

  • Taylor, R.B., Duffus, W.P.H., Raff, M.C., and de Petris, S.: Redistribution and pinocytosis of lymphocyte surface immunoglobulin molecules induced by anti-immunoglobulin antibody.Nature New Biol. 233:225–229, 1971

    Google Scholar 

  • Unanue, E.R., Dorf, E.M., David, C.S., and Benacerraf, B.: The presence ofI-region-associated antigens on B cells in molecules distinct from immunoglobulin and H-2K and H-2D.Proc. Natl. Acad. Sci. USA 71:5014–5016, 1974

    Google Scholar 

  • Wagner, H.: Synergy during in vitro cytotoxic allograft responses: I. Evidence for cell interactions between thymocytes and peripheral T cells.J. Exp. Med. 138:1379–1397, 1973

    Google Scholar 

  • Wagner, H., Röllinghoff, M., and Nossal, G.J.V.: T-cell mediated immune responses induced in vitro: A probe for allograft and tumor immunity.Transplant. Rev. 17:3–36, 1973

    Google Scholar 

  • Wagner, H., Röllinghoff, M., and Shortmann, K.: Evidence for T-T cell synergism during in vitro cytotoxic allograft responses.In L. Brent and J. Holborow (eds.):Progress in Immunology II, Vol. 3; pp. 111–120, North-Holland Publ. Co., Amsterdam, 1974

    Google Scholar 

  • Wagner, H. and Wyss, C.: Cell-mediated immune response in vitro: V. A comparative study of in vitro immunogenicity of splenic lymphocytes, neoplastic lymphoid cells and fibroblasts.Eur. J. Immunol. 3:549–555, 1973

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Medizinische Mikrobiologie, Hochhaus am Augustusplatz, 65 Mainz, Germany

    Hermann Wagner & Martin Röllinghoff

  2. Institut für Genetik, 5 Köln-Lindenthal, Germany

    Günther Hämmerling

Authors
  1. Hermann Wagner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Günther Hämmerling
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martin Röllinghoff
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wagner, H., Hämmerling, G. & Röllinghoff, M. Enhanced in vitro cytotoxic anti H-2 responses in the presence of Ia antigens. Immunogenetics 2, 257–268 (1975). https://doi.org/10.1007/BF01572294

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01572294

Keywords

Search

Navigation