Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1981 Jan 1;153(1):107–128. doi: 10.1084/jem.153.1.107

In vitro analysis of allogeneic lymphocyte interaction. V. Identification and characterization of two components of allogeneic effect factor, one of which displays H-2-restricted helper activity and the other, T cell-growth factor activity

PMCID: PMC2186045  PMID: 6969778

Abstract

An allogeneic effect factor (AEF) derived from mixed lymphocyte reaction (MLR) cultures of alloactivated A.SW (H-2s) responder T cells and irradiated A/WySn (H-2a) stimulator spleen cells helps an in vitro primary anti-erythrocyte plaque-forming cell PFC response of BALB/c nude spleen cels and also A/WySn but not A.SW T cell-depleted spleen cells. AEF activity is adsorbed by anti-Ik and anti-I-Ak but not by anti-I-Jk, anti-I-ECk, and anti-Is. Gel filtration of ACA 54 resolves AEF into two main components that which appear in the 50,000- to 70,000- mol wt (component I) and 30,000- to 35,000-mol wt (component II) regions, respectively. Component I has a mol wt of 68,000, elutes from DEAE-Sephacel at 0.05-0.1 M NaCl, and has an isoelectric point (pI) of 5.8. It helps A/WySn but not A.SW B cells and, therefore, is H-2 restricted. Component II is not H-2 restricted, because it helps both A.SW and A/WySn B cells. It also stimulates (a) the growth of a long- term cytotoxic cell line in vitro, (b) Con A-induced thymocyte mitogenesis, and (c) the generation of cytotoxic T cells. The latter three properties of component II are not shared by component I. In addition, component II elutes from DEAE-Sephacel at 0.15-0.2 M NaCl and has a pI of 4.3 and 4.9. Ia determinants and Ig VH, CH, L-chain, and idiotypic determinants are not present on either component I or component II. The properties of component II are identical to that of a T cell growth factor produced by Con A-stimulated spleen cells. It is suggested that the H-2-restricted component I of AEF might be an MLR- activated responder T cell-derived Ia alloantigen receptor.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

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

  1. Altman A., Cardenas J. M., Bechtold T. E., Katz D. H. The biologic effects of allogeneic effect factor on T lymphocytes. I. The mitogenic activity and the autonomous induction of cytotoxic T lymphocytes by AEF. J Immunol. 1980 Jan;124(1):105–113. [PubMed] [Google Scholar]
  2. Armerding D., Eshhar Z., Katz D. H. Activation of T and B lymphocytes in vitro. VI. Biochemical and physicochemical characterization of the allogeneic effect factor. J Immunol. 1977 Oct;119(4):1468–1474. [PubMed] [Google Scholar]
  3. Binz H., Wigzell H. Shared idiotypic determinants on B and T lymphocytes reactive against the same antigenic determinants. V. Biochemical and serological characteristics of naturally occurring, soluble antigen-binding T-lymphocyte-derived molecules. Scand J Immunol. 1976;5(5):559–571. doi: 10.1111/j.1365-3083.1976.tb00311.x. [DOI] [PubMed] [Google Scholar]
  4. Cantor H., Boyse E. A. Lymphocytes as models for the study of mammalian cellular differentiation. Immunol Rev. 1977 Jan;33:105–124. doi: 10.1111/j.1600-065x.1977.tb00364.x. [DOI] [PubMed] [Google Scholar]
  5. Cook R. G., Vitetta E. S., Uhr J. W., Capra J. D. Structural studies on the murine Ia alloantigens--III. Tryptic peptide comparisons of allelic products of the I-E/C sub-region. Mol Immunol. 1979 Jan;16(1):29–35. doi: 10.1016/0161-5890(79)90024-5. [DOI] [PubMed] [Google Scholar]
  6. Delovitch T. L., Biggin J., Fung F. Y. In vitro analysis of allogeneic lymphocyte interaction. II. I-region control of the activity of a B-cell-derived H-2-restricted allogeneic effect factor and its receptor during B-cell activation. J Exp Med. 1978 Apr 1;147(4):1198–1212. doi: 10.1084/jem.147.4.1198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Delovitch T. L., Fegelman A., Barber B. H., Frelinger J. A. Immunochemical characterization of the Ly-8.2 murine lymphocyte alloantigen: possible relationship to actin. J Immunol. 1979 Jan;122(1):326–333. [PubMed] [Google Scholar]
  8. Delovitch T. L., McDevitt H. O. In vitro analysis of allogeneic lymphocyte interaction. I. Characterization and cellular origin of an Ia-positive helper factor-allogeneic effect factor. J Exp Med. 1977 Oct 1;146(4):1019–1032. doi: 10.1084/jem.146.4.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Delovitch T. L., Sohn U. In vitro analysis of allogeneic lymphocyte interaction. IV. Dual recognition of B cell-associated Mls locus and I-region determinants by a helper allogeneic effect factor (AEF) generated across a minor H locus disparity. J Immunol. 1979 Jul;123(1):121–127. [PubMed] [Google Scholar]
  10. Fathman C. G., Hengartner H. Crossreactive mixed lymphocyte reaction determinants recognized by cloned alloreactive T cells. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5863–5866. doi: 10.1073/pnas.76.11.5863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gillis S., Smith K. A., Watson J. Biochemical characterization of lymphocyte regulatory molecules. II. Purification of a class of rat and human lymphokines. J Immunol. 1980 Apr;124(4):1954–1962. [PubMed] [Google Scholar]
  12. Harris J. F., Delovitch T. L. Derivation of a monoclonal antibody that detects an Ia antigen encoded by 2 complementing I-subregions. J Immunol. 1980 Nov;125(5):2167–2176. [PubMed] [Google Scholar]
  13. Jones P. P., Murphy D. B., McDevitt H. O. Two-gene control of the expression of a murine Ia antigen. J Exp Med. 1978 Oct 1;148(4):925–939. doi: 10.1084/jem.148.4.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Langone J. J., Boyle M. D., Borsos T. 125I protein A: applications to the quantitative determination of fluid phase and cell-bound IgG. J Immunol Methods. 1977;18(3-4):281–293. doi: 10.1016/0022-1759(77)90182-x. [DOI] [PubMed] [Google Scholar]
  15. Larsson E. L., Iscove N. N., Coutinho A. Two distinct factors are required for induction of T-cell growth. Nature. 1980 Feb 14;283(5748):664–666. doi: 10.1038/283664a0. [DOI] [PubMed] [Google Scholar]
  16. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  17. Marshak-Rothstein A., Fink P., Gridley T., Raulet D. H., Bevan M. J., Gefter M. L. Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. J Immunol. 1979 Jun;122(6):2491–2497. [PubMed] [Google Scholar]
  18. Murray M. J., Kabat D. Genetic and sialylation sources of heterogeneity of the murine leukemia virus membrane envelope glycoproteins gp69/71. J Biol Chem. 1979 Feb 25;254(4):1340–1348. [PubMed] [Google Scholar]
  19. Nagy Z., Elliott B. E., Nabholz M. Specific binding of K- and I-region products of the H-2 complex to activated thymus-derived (T) cells belonging to different Ly subclasses. J Exp Med. 1976 Dec 1;144(6):1545–1553. doi: 10.1084/jem.144.6.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  21. Pacifico A., Capra J. D. T cell hybrids with arsonate specificity. I. Initial characterization of antigen-specific T cell products that bear a cross-reactive idiotype and determinants encoded by the murine major histocompatibility complex. J Exp Med. 1980 Nov 1;152(5):1289–1301. doi: 10.1084/jem.152.5.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Paetkau V., Shaw J., Mills G., Caplan B. Cellular origins and targets of costimulator (IL2). Immunol Rev. 1980;51:157–175. doi: 10.1111/j.1600-065x.1980.tb00320.x. [DOI] [PubMed] [Google Scholar]
  23. Paul W. E., Benacerraf B. Functional specificity of thymus- dependent lymphocytes. Science. 1977 Mar 25;195(4284):1293–1300. doi: 10.1126/science.320663. [DOI] [PubMed] [Google Scholar]
  24. Poole T., Leach B. S., Fish W. W. Analysis of polypeptide molecular weights by electrophoresis in urea. Anal Biochem. 1974 Aug;60(2):596–607. doi: 10.1016/0003-2697(74)90272-3. [DOI] [PubMed] [Google Scholar]
  25. Prud'homme G. J., Sohn U., Delovitch T. L. The role of H-2 and Ia antigens in graft-versus-host reactions (GVHR). Presence of host alloantigens on donor cells after GVHR and suppression of GVHR with an anti-Ia antiserum against hose Ia antigens. J Exp Med. 1979 Jan 1;149(1):137–149. doi: 10.1084/jem.149.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schrader J. W., Arnold B., Clark-Lewis I. A con A-stimulated T-cell hybridoma releases factors affecting haematopoietic colony-forming cells and B-cell antibody responses. Nature. 1980 Jan 10;283(5743):197–199. doi: 10.1038/283197a0. [DOI] [PubMed] [Google Scholar]
  27. Schwartz R. H., Chen C., Paul W. E. Gene complementation in the T lymphocyte proliferative response to poly (Glu56Lys35Phe9)n. Functional evidence for a restriction element coded for by both the I-A and I-E subregions. Eur J Immunol. 1980 Sep;10(9):708–714. doi: 10.1002/eji.1830100910. [DOI] [PubMed] [Google Scholar]
  28. Shaw J., Caplan B., Paetkau V., Pilarski L. M., Delovitch T. L., McKenzie I. F. Cellular origins of co-stimulator (IL2) and its activity in cytotoxic T lymphocyte responses. J Immunol. 1980 May;124(5):2231–2239. [PubMed] [Google Scholar]
  29. Shaw J., Monticone V., Paetkau V. Partial purification and molecular characterization of a lymphokine (costimulator) required for the mitogenic response of mouse thymocytes in vitro. J Immunol. 1978 Jun;120(6):1967–1973. [PubMed] [Google Scholar]
  30. Singer A., Hathcock K. S., Hodes R. J. Cellular and genetic control of antibody responses. V. Helper T-cell recognition of H-2 determinants on accessory cells but not B cells. J Exp Med. 1979 May 1;149(5):1208–1226. doi: 10.1084/jem.149.5.1208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sprent J. Role of H-2 gene products in the function of T helper cells from normal and chimeric mice in vivo. Immunol Rev. 1978;42:108–137. doi: 10.1111/j.1600-065x.1978.tb00260.x. [DOI] [PubMed] [Google Scholar]
  32. Taniguchi M., Takei I., Tada T. Functional and molecular organisation of an antigen-specific suppressor factor from a T-cell hybridoma. Nature. 1980 Jan 10;283(5743):227–228. doi: 10.1038/283227a0. [DOI] [PubMed] [Google Scholar]
  33. Watson J., Gillis S., Marbrook J., Mochizuki D., Smith K. A. Biochemical and biological characterization of lymphocyte regulatory molecules. I. Purification of a class of murine lymphokines. J Exp Med. 1979 Oct 1;150(4):849–861. doi: 10.1084/jem.150.4.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zinkernagel R. M. Thymus and lymphohemopoietic cells: their role in T cell maturation in selection of T cells' H-2-restriction-specificity and in H-2 linked Ir gene control. Immunol Rev. 1978;42:224–270. doi: 10.1111/j.1600-065x.1978.tb00264.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES