Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1985 May 1;161(5):953–971. doi: 10.1084/jem.161.5.953

Obligatory role of gamma interferon in T cell-replacing factor- dependent, antigen-specific murine B cell responses

PMCID: PMC2187618  PMID: 2580939

Abstract

The role of gamma interferon (IFN-gamma) in T cell-replacing factor (TRF) activity for antigen-specific plaque-forming cell (PFC) responses in vitro was studied using antibodies to murine IFN-gamma (Mu IFN- gamma). TRF activity was present in supernatants (Sn) of Con A- or mixed leukocyte reaction-stimulated murine spleen cells as well as in an IL-2-rich fraction of phytohemagglutinin-stimulated human peripheral blood lymphocyte Sn and in the Sn of the Gibbon T lymphoma MLA-144. The human TRF was highly active with cells from nu/nu mice and normal mice but not with cells from animals with the xid immunologic defect, similar to the activity of murine TRF. Antibodies to IFN-gamma consisted of hyper-immune rabbit antisera, IFN-gamma affinity-purified rabbit immunoglobulin and an interspecies hybridoma specific for Mu IFN- gamma. The results show that the activities of all preparations of TRF are markedly diminished or abrogated by antibody to Mu IFN-gamma but not by antibodies to human IFN-gamma (Hu IFN-gamma), nor by normal rabbit sera or purified rabbit Ig. The degree of inhibition was dose dependent and was quantitatively reversed by the addition to the cultures of recombinant-derived Mu IFN-gamma (Mu rIFN-gamma) but not Hu rIFN-gamma. This reversal was fully antigen specific and thus not attributable to polyclonal B cell activation by IFN-gamma, which is inactive alone in the TRF assay. Kinetic analysis shows that IFN-gamma must act by 24-48 h to produce PFC responses at 4 d. Together, the data demonstrate that IFN-gamma is a necessary mediator for TRF effects and that IFN-gamma is induced by TRF from T-depleted murine spleen cells in sufficient quantity to support large antibody responses. The source of this IFN-gamma may be the potent natural killer cells that are induced in cultures stimulated with TRF.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. 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]
  2. Cunningham A. J., Szenberg A. Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology. 1968 Apr;14(4):599–600. [PMC free article] [PubMed] [Google Scholar]
  3. Eisenberg L., Prystowsky M. B., Dick R. F., Sosman J. A., Fitch F. W., Quintáns J. TRF requirements for in vitro PFC responses to SRBC and R36a. I. TRF is distinct from IL 2 but indistinguishable from polyclonal BCSF. J Immunol. 1984 Mar;132(3):1305–1310. [PubMed] [Google Scholar]
  4. Endres R. O., Kushnir E., Kappler J. W., Marrack P., Kinsky S. C. A requirement for nonspecific T cell factors in antibody responses to "T cell independent" antigens. J Immunol. 1983 Feb;130(2):781–784. [PubMed] [Google Scholar]
  5. Farrar J. J. The xenogeneic effect. I. Antigen and mitogen-stimulated human lymphocytes produce a non-antigen-specific factor which reconstitutes the antibody response of T cell-deficient mouse spleen cells. J Immunol. 1975 Nov;115(5):1295–1300. [PubMed] [Google Scholar]
  6. Farrar W. L., Johnson H. M., Farrar J. J. Regulation of the production of immune interferon and cytotoxic T lymphocytes by interleukin 2. J Immunol. 1981 Mar;126(3):1120–1125. [PubMed] [Google Scholar]
  7. Gillis S., Union N. A., Baker P. E., Smith K. A. The in vitro generation and sustained culture of nude mouse cytolytic T-lymphocytes. J Exp Med. 1979 Jun 1;149(6):1460–1476. doi: 10.1084/jem.149.6.1460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gray P. W., Goeddel D. V. Cloning and expression of murine immune interferon cDNA. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5842–5846. doi: 10.1073/pnas.80.19.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gray P. W., Leung D. W., Pennica D., Yelverton E., Najarian R., Simonsen C. C., Derynck R., Sherwood P. J., Wallace D. M., Berger S. L. Expression of human immune interferon cDNA in E. coli and monkey cells. Nature. 1982 Feb 11;295(5849):503–508. doi: 10.1038/295503a0. [DOI] [PubMed] [Google Scholar]
  10. Greenstein J. L., Lord E., Kappler J. W., Marrack P. C. Analysis of the response of B cells from CBA/N-defective mice to nonspecific T cell help. J Exp Med. 1981 Nov 1;154(5):1608–1617. doi: 10.1084/jem.154.5.1608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Handa K., Suzuki R., Matsui H., Shimizu Y., Kumagai K. Natural killer (NK) cells as a responder to interleukin 2 (IL 2). II. IL 2-induced interferon gamma production. J Immunol. 1983 Feb;130(2):988–992. [PubMed] [Google Scholar]
  12. Howard M., Matis L., Malek T. R., Shevach E., Kell W., Cohen D., Nakanishi K., Paul W. E. Interleukin 2 induces antigen-reactive T cell lines to secrete BCGF-I. J Exp Med. 1983 Dec 1;158(6):2024–2039. doi: 10.1084/jem.158.6.2024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Howard M., Paul W. E. Regulation of B-cell growth and differentiation by soluble factors. Annu Rev Immunol. 1983;1:307–333. doi: 10.1146/annurev.iy.01.040183.001515. [DOI] [PubMed] [Google Scholar]
  14. Kawase I., Brooks C. G., Kuribayashi K., Olabuenaga S., Newman W., Gillis S., Henney C. S. Interleukin 2 induces gamma-interferon production: participation of macrophages and NK-like cells. J Immunol. 1983 Jul;131(1):288–292. [PubMed] [Google Scholar]
  15. Kleinschmidt W. J., Schultz R. M. Similarities of murine gamma interferon and the lymphokine that renders macrophages cytotoxic. J Interferon Res. 1982;2(2):291–299. doi: 10.1089/jir.1982.2.291. [DOI] [PubMed] [Google Scholar]
  16. Leibson H. J., Gefter M., Zlotnik A., Marrack P., Kappler J. W. Role of gamma-interferon in antibody-producing responses. 1984 Jun 28-Jul 4Nature. 309(5971):799–801. doi: 10.1038/309799a0. [DOI] [PubMed] [Google Scholar]
  17. Leibson H. J., Marrack P., Kappler J. W. B cell helper factors. I. Requirement for both interleukin 2 and another 40,000 mol wt factor. J Exp Med. 1981 Nov 1;154(5):1681–1693. doi: 10.1084/jem.154.5.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liebson H. J., Marrack P., Kappler J. B cell helper factors. II. Synergy among three helper factors in the response of T cell- and macrophage-depleted B cells. J Immunol. 1982 Oct;129(4):1398–1402. [PubMed] [Google Scholar]
  19. Ly I. A., Mishell R. I. Separation of mouse spleen cells by passage through columns of sephadex G-10. J Immunol Methods. 1974 Aug;5(3):239–247. doi: 10.1016/0022-1759(74)90108-2. [DOI] [PubMed] [Google Scholar]
  20. Muraguchi A., Kasahara T., Oppenheim J. J., Fauci A. S. B cell growth factor and T cell growth factor produced by mitogen-stimulated normal human peripheral blood T lymphocytes are distinct molecules. J Immunol. 1982 Dec;129(6):2486–2489. [PubMed] [Google Scholar]
  21. Parker D. C. Induction and suppression of polyclonal antibody responses by anti-Ig reagents and antigen-nonspecific helper factors: a comparison of the effects of anti-Fab, anti-IgM, and anti IgD on murine B cells. Immunol Rev. 1980;52:115–139. doi: 10.1111/j.1600-065x.1980.tb00333.x. [DOI] [PubMed] [Google Scholar]
  22. Parker D. C. Separable helper factors support B cell proliferation and maturation to Ig secretion. J Immunol. 1982 Aug;129(2):469–474. [PubMed] [Google Scholar]
  23. Ralph P., Welte K., Levi E., Nakoinz I., Litcofsky P. B., Mertelsmann R. H., Moore M. A. Human B cell-inducing factor(s) for production of IgM, IgG and IgA: independence from IL 2. J Immunol. 1984 Apr;132(4):1858–1862. [PubMed] [Google Scholar]
  24. Roehm N. W., Marrack P., Kappler J. W. Helper signals in the plaque-forming cell response to protein-bound haptens. J Exp Med. 1983 Aug 1;158(2):317–333. doi: 10.1084/jem.158.2.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schimpl A., Wecker E. A third signal in B cell activation given by TRF. Transplant Rev. 1975;23:176–188. doi: 10.1111/j.1600-065x.1975.tb00157.x. [DOI] [PubMed] [Google Scholar]
  26. Sidman C. L., Marshall J. D., Shultz L. D., Gray P. W., Johnson H. M. Gamma-interferon is one of several direct B cell-maturing lymphokines. 1984 Jun 28-Jul 4Nature. 309(5971):801–804. doi: 10.1038/309801a0. [DOI] [PubMed] [Google Scholar]
  27. Simon P. L., Farrar J. J., Dind P. D. Biochemical relationship between murine immune interferon and a killer cell helper factor. J Immunol. 1979 Jan;122(1):127–132. [PubMed] [Google Scholar]
  28. Singer A., Asano Y., Shigeta M., Hathcock K. S., Ahmed A., Fathman C. G., Hodes R. J. Distinct B cell subpopulations differ in their genetic requirements for activation by T helper cells. Immunol Rev. 1982;64:137–160. doi: 10.1111/j.1600-065x.1982.tb00422.x. [DOI] [PubMed] [Google Scholar]
  29. Spitalny G. L., Havell E. A. Monoclonal antibody to murine gamma interferon inhibits lymphokine-induced antiviral and macrophage tumoricidal activities. J Exp Med. 1984 May 1;159(5):1560–1565. doi: 10.1084/jem.159.5.1560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Suzuki R., Handa K., Itoh K., Kumagai K. Natural killer (NK) cells as a responder to interleukin 2 (IL 2). I. Proliferative response and establishment of cloned cells. J Immunol. 1983 Feb;130(2):981–987. [PubMed] [Google Scholar]
  31. Svedersky L. P., Benton C. V., Berger W. H., Rinderknecht E., Harkins R. N., Palladino M. A. Biological and antigenic similarities of murine interferon-gamma and macrophage-activating factor. J Exp Med. 1984 Mar 1;159(3):812–827. doi: 10.1084/jem.159.3.812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Swain S. L., Dennert G., Warner J. F., Dutton R. W. Culture supernatants of a stimulated T-cell line have helper activity that acts synergistically with interleukin 2 in the response of B cells to antigen. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2517–2521. doi: 10.1073/pnas.78.4.2517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Swain S. L., Dutton R. W. Production of Con A-induced helper T cell replacing factor requires a T cell and an Ia-positive non-T cells. J Immunol. 1980 Jan;124(1):437–444. [PubMed] [Google Scholar]
  34. Swain S. L., Dutton R. W. Production of a B cell growth-promoting activity, (DL)BCGF, from a cloned T cell line and its assay on the BCL1 B cell tumor. J Exp Med. 1982 Dec 1;156(6):1821–1834. doi: 10.1084/jem.156.6.1821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Swain S. L., Howard M., Kappler J., Marrack P., Watson J., Booth R., Wetzel G. D., Dutton R. W. Evidence for two distinct classes of murine B cell growth factors with activities in different functional assays. J Exp Med. 1983 Sep 1;158(3):822–835. doi: 10.1084/jem.158.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Swain S. L., Wetzel G. D., Soubiran P., Dutton R. W. T cell replacing factors in the B cell response to antigen. Immunol Rev. 1982;63:111–128. doi: 10.1111/j.1600-065x.1982.tb00413.x. [DOI] [PubMed] [Google Scholar]
  37. Walker E. B., Maino V., Sanchez-Lanier M., Warner N., Stewart C. Murine gamma interferon activates the release of a macrophage-derived Ia-inducing factor that transfers Ia inductive capacity. J Exp Med. 1984 May 1;159(5):1532–1547. doi: 10.1084/jem.159.5.1532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wong G. H., Clark-Lewis I., McKimm-Breschkin L., Harris A. W., Schrader J. W. Interferon-gamma induces enhanced expression of Ia and H-2 antigens on B lymphoid, macrophage, and myeloid cell lines. J Immunol. 1983 Aug;131(2):788–793. [PubMed] [Google Scholar]
  39. Yamamoto J. K., Farrar W. L., Johnson H. M. Interleukin 2 regulation of mitogen induction of immune interferon (IFN gamma) in spleen cells and thymocytes. Cell Immunol. 1982 Jan 15;66(2):333–341. doi: 10.1016/0008-8749(82)90183-6. [DOI] [PubMed] [Google Scholar]
  40. Zlotnik A., Roberts W. K., Vasil A., Blumenthal E., Larosa F., Leibson H. J., Endres R. O., Graham S. D., Jr, White J., Hill J. Coordinate production by a T cell hybridoma of gamma interferon and three other lymphokine activities: multiple activities of a single lymphokine? J Immunol. 1983 Aug;131(2):794–800. [PubMed] [Google Scholar]

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

RESOURCES