Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Apr 1;167(4):1377–1390. doi: 10.1084/jem.167.4.1377

Recombinant interleukin 2 or 5, but not 3 or 4, induces maturation of resting mouse B lymphocytes and propagates proliferation of activated B cell blasts

PMCID: PMC2188910  PMID: 3128631

Abstract

Plasmacytoma transformants of the X63-Ag8-653 cell line carrying an expression vector with either IL-2, -3, -4, or -5 cDNA were established that secrete the corresponding ILs at high rates. The four mouse ILs (mILs) were then tested as single ILs and in combinations for their effects on the maturation of resting and proliferation of activated normal mouse splenic B cells. mIL-3 and mIL-4 were inactive in all assays. mIL-2, as well as mIL-5, synergized with Ig-specific antibodies and B cell growth factor alpha (BCGF-alpha) to stimulate successive rounds of B cell division with LPS-activated B cells. This activity as BCGF-beta was effective at concentrations similar to those at which mIL- 2 induced proliferation of the CTL-L T cell line, indicating a high- affinity interaction of both mIL-2 and mIL-5 with their corresponding receptors on activated B cells. mIL-5 and maybe IL-2 also induced maturation of resting B cells to Ig-secreting cells without proliferation. This B cell maturation factor (BMF) activity of mIL-5 was as effective as its BCGF-beta activity, while the BMF activity of mIL-2 was at least 10(2)-fold less effective. BMF activity of mIL-2, but not mIL-5, was blocked by anti-Il-2-R antibodies, indicating that mIL-2 and mIL-5 use separate receptors for B cell signaling. mIL-2, as well as mIL-5, furthermore, acted as filler activities when proliferation in the presence of Ig-specific antibodies and BCGF-alpha was measured with as little as 500 B cells. In the case of mIL-5, this was also true for maturation of that few cells. Limiting dilution analyses showed that approximately 1-2% of the resting B cells matured without division, while 30-100-fold fewer cells (0.03-0.06%) proliferated and matured in response to IL-5. A single IL, therefore, is capable of inducing maturation and of stimulating mitotic cell cycle progression of normal B cells.

Full Text

The Full Text of this article is available as a PDF (987.8 KB).

Selected References

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

  1. Andersson J., Coutinho A., Lernhardt W., Melchers F. Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte. Cell. 1977 Jan;10(1):27–34. doi: 10.1016/0092-8674(77)90136-2. [DOI] [PubMed] [Google Scholar]
  2. Andersson J., Schreier M. H., Melchers F. T-cell-dependent B-cell stimulation is H-2 restricted and antigen dependent only at the resting B-cell level. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1612–1616. doi: 10.1073/pnas.77.3.1612. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Coffman R. L., Ohara J., Bond M. W., Carty J., Zlotnik A., Paul W. E. B cell stimulatory factor-1 enhances the IgE response of lipopolysaccharide-activated B cells. J Immunol. 1986 Jun 15;136(12):4538–4541. [PubMed] [Google Scholar]
  4. Corbel C., Melchers F. The synergism of accessory cells and of soluble alpha-factors derived from them in the activation of B cells to proliferation. Immunol Rev. 1984 Apr;78:51–74. doi: 10.1111/j.1600-065x.1984.tb00476.x. [DOI] [PubMed] [Google Scholar]
  5. Gronowicz E., Coutinho A., Melchers F. A plaque assay for all cells secreting Ig of a given type or class. Eur J Immunol. 1976 Aug;6(8):588–590. doi: 10.1002/eji.1830060812. [DOI] [PubMed] [Google Scholar]
  6. Harada N., Kikuchi Y., Tominaga A., Takaki S., Takatsu K. BCGFII activity on activated B cells of a purified murine T cell-replacing factor (TRF) from a T cell hybridoma (B151K12). J Immunol. 1985 Jun;134(6):3944–3951. [PubMed] [Google Scholar]
  7. 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]
  8. Iscove N. N., Melchers F. Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes. J Exp Med. 1978 Mar 1;147(3):923–933. doi: 10.1084/jem.147.3.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Leptin M. Monoclonal antibodies specific for murine IgM. II. Activation of B lymphocytes by monoclonal antibodies specific for the four constant domains of IgM. Eur J Immunol. 1985 Feb;15(2):131–137. doi: 10.1002/eji.1830150206. [DOI] [PubMed] [Google Scholar]
  10. Leptin M., Potash M. J., Grützmann R., Heusser C., Shulman M., Köhler G., Melchers F. Monoclonal antibodies specific for murine IgM I. Characterization of antigenic determinants on the four constant domains of the mu heavy chain. Eur J Immunol. 1984 Jun;14(6):534–542. doi: 10.1002/eji.1830140610. [DOI] [PubMed] [Google Scholar]
  11. Lernhardt W., Andersson J., Coutinho A., Melchers F. Cloning of murine transformed cell lines in suspension culture with efficiencies near 100%. Exp Cell Res. 1978 Feb;111(2):309–316. doi: 10.1016/0014-4827(78)90175-1. [DOI] [PubMed] [Google Scholar]
  12. Lowenthal J. W., Zubler R. H., Nabholz M., MacDonald H. R. Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes. Nature. 1985 Jun 20;315(6021):669–672. doi: 10.1038/315669a0. [DOI] [PubMed] [Google Scholar]
  13. Malek T. R., Robb R. J., Shevach E. M. Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5694–5698. doi: 10.1073/pnas.80.18.5694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Melchers F., Andersson J. Synthesis, surface deposition and secretion of immunoglobulin M in bone marrow-derived lymphocytes before and after mitogenic stimulation. Transplant Rev. 1973;14:76–130. doi: 10.1111/j.1600-065x.1973.tb00103.x. [DOI] [PubMed] [Google Scholar]
  15. Melchers F., Lernhardt W. Three restriction points in the cell cycle of activated murine B lymphocytes. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7681–7685. doi: 10.1073/pnas.82.22.7681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller R. G., Phillips R. A. Separation of cells by velocity sedimentation. J Cell Physiol. 1969 Jun;73(3):191–201. doi: 10.1002/jcp.1040730305. [DOI] [PubMed] [Google Scholar]
  17. Nakagawa T., Hirano T., Nakagawa N., Yoshizaki K., Kishimoto T. Effect of recombinant IL 2 and gamma-IFN on proliferation and differentiation of human B cells. J Immunol. 1985 Feb;134(2):959–966. [PubMed] [Google Scholar]
  18. Nakanishi K., Howard M., Muraguchi A., Farrar J., Takatsu K., Hamaoka T., Paul W. E. Soluble factors involved in B cell differentiation: identification of two distinct T cell-replacing factors (TRF). J Immunol. 1983 May;130(5):2219–2224. [PubMed] [Google Scholar]
  19. Nakanishi K., Malek T. R., Smith K. A., Hamaoka T., Shevach E. M., Paul W. E. Both interleukin 2 and a second T cell-derived factor in EL-4 supernatant have activity as differentiation factors in IgM synthesis. J Exp Med. 1984 Dec 1;160(6):1605–1621. doi: 10.1084/jem.160.6.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Noelle R., Krammer P. H., Ohara J., Uhr J. W., Vitetta E. S. Increased expression of Ia antigens on resting B cells: an additional role for B-cell growth factor. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6149–6153. doi: 10.1073/pnas.81.19.6149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pike B. L., Raubitschek A., Nossal G. J. Human interleukin 2 can promote the growth and differentiation of single hapten-specific B cells in the presence of specific antigen. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7917–7921. doi: 10.1073/pnas.81.24.7917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Riendeau D., Harnish D. G., Bleackley R. C., Paetkau V. Purification of mouse interleukin 2 to apparent homogeneity. J Biol Chem. 1983 Oct 25;258(20):12114–12117. [PubMed] [Google Scholar]
  23. Roehm N. W., Leibson H. J., Zlotnik A., Kappler J., Marrack P., Cambier J. C. Interleukin-induced increase in Ia expression by normal mouse B cells. J Exp Med. 1984 Sep 1;160(3):679–694. doi: 10.1084/jem.160.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sideras P., Bergstedt-Lindqvist S., MacDonald H. R., Severinson E. Secretion of IgG1 induction factor by T cell clones and hybridomas. Eur J Immunol. 1985 Jun;15(6):586–593. doi: 10.1002/eji.1830150611. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Takatsu K., Harada N., Hara Y., Takahama Y., Yamada G., Dobashi K., Hamaoka T. Purification and physicochemical characterization of murine T cell replacing factor (TRF). J Immunol. 1985 Jan;134(1):382–389. [PubMed] [Google Scholar]
  28. Vitetta E. S., Brooks K., Chen Y. W., Isakson P., Jones S., Layton J., Mishra G. C., Pure E., Weiss E., Word C. T-cell-derived lymphokines that induce IgM and IgG secretion in activated murine B cells. Immunol Rev. 1984 Apr;78:137–157. doi: 10.1111/j.1600-065x.1984.tb00480.x. [DOI] [PubMed] [Google Scholar]
  29. Vitetta E. S., Ohara J., Myers C. D., Layton J. E., Krammer P. H., Paul W. E. Serological, biochemical, and functional identity of B cell-stimulatory factor 1 and B cell differentiation factor for IgG1. J Exp Med. 1985 Nov 1;162(5):1726–1731. doi: 10.1084/jem.162.5.1726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zubler R. H., Lowenthal J. W., Erard F., Hashimoto N., Devos R., MacDonald H. R. Activated B cells express receptors for, and proliferate in response to, pure interleukin 2. J Exp Med. 1984 Oct 1;160(4):1170–1183. doi: 10.1084/jem.160.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES