Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1987 Dec 1;6(12):3687–3693. doi: 10.1002/j.1460-2075.1987.tb02702.x

Ly1+ PRO-B lymphocyte clones. Phenotype, growth requirements and differentiation in vitro and in vivo.

R Palacios 1, H Karasuyama 1, A Rolink 1
PMCID: PMC553838  PMID: 3501371

Abstract

Several clones obtained from the bone marrow of a BALB/c mouse were found to contain the heavy and light chain Ig genes in the germline configuration, to express Ly1 and to carry the B cell lineage markers B-220, Lyb8 and BP-1; these clones are Pgp-1+, LFA-1+, J11d+, Mac-1+ and Thy1-, Lyt2-, L3T4-, GM1.2- and Ia-. Three clones analyzed in detail (Lyd9, LyH7 and Lyb9) have receptors for interleukin (IL) 2 and IL3 as assessed with the 7D4 and CC11 monoclonal antibodies respectively. They grow in rIL3 but not in rIL2 or rIL1; both rIL4 and rIL5 also promote their proliferation, albeit to a much lesser extent than rIL3. None of the interleukins tested alone or in various combinations promoted the clones to differentiate in vitro along the B cell pathway. Treatment with 5-Azacytidine (5-Aza) induced cell surface Ia expression but not rearrangement or expression of Ig genes. However, 5-Aza-treated Lyd9, LyH7 and Lyb9 cells co-cultured with X-ray irradiated accessory cells and LPS gave rise to Ly1+, IgM+ B lymphocytes (range 14-51%) including mu + kappa + (78-93%), and mu + lambda + (9-25%) B lymphocytes. In vivo, the Lyd9, LyH7 and Lyb9 clones gave rise to IgM+ B lymphocytes (8.5-17%) including mu + kappa +, and mu + lambda +, but not to Lyt2+ or L3T4+ T lymphocytes after 4-6 weeks of transfer into Scid mice. Our results indicate that Ly1+ IgM+ cells comprise a subpopulation of B lymphocytes that is derived from IL3-responsive Ly1+ PRO-B lymphocytes.

Full text

PDF
3689

Images in this article

Selected References

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

  1. Alt F. W., Blackwell T. K., DePinho R. A., Reth M. G., Yancopoulos G. D. Regulation of genome rearrangement events during lymphocyte differentiation. Immunol Rev. 1986 Feb;89:5–30. doi: 10.1111/j.1600-065x.1986.tb01470.x. [DOI] [PubMed] [Google Scholar]
  2. Bosma G. C., Custer R. P., Bosma M. J. A severe combined immunodeficiency mutation in the mouse. Nature. 1983 Feb 10;301(5900):527–530. doi: 10.1038/301527a0. [DOI] [PubMed] [Google Scholar]
  3. Braun J., Citri Y., Baltimore D., Forouzanpour F., King L., Teheranizadeh K., Bray M., Kliewer S. B-Ly1 cells: immortal Ly-1+ B lymphocyte cell lines spontaneously arising in murine splenic cultures. Immunol Rev. 1986 Oct;93:5–21. doi: 10.1111/j.1600-065x.1986.tb01499.x. [DOI] [PubMed] [Google Scholar]
  4. Cooper M. D., Mulvaney D., Coutinho A., Cazenave P. A. A novel cell surface molecule on early B-lineage cells. Nature. 1986 Jun 5;321(6070):616–618. doi: 10.1038/321616a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cory S., Adams J. M., Kemp D. J. Somatic rearrangements forming active immunoglobulin mu genes in B and T lymphoid cell lines. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4943–4947. doi: 10.1073/pnas.77.8.4943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dasch J. R., Jones P. P. Independent regulation of IgM, IgD, and Ia antigen expression in cultured immature B lymphocytes. J Exp Med. 1986 Apr 1;163(4):938–951. doi: 10.1084/jem.163.4.938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davidson W. F., Fredrickson T. N., Rudikoff E. K., Coffman R. L., Hartley J. W., Morse H. C., 3rd A unique series of lymphomas related to the Ly-1+ lineage of B lymphocyte differentiation. J Immunol. 1984 Aug;133(2):744–753. [PubMed] [Google Scholar]
  9. Dorshkind K., Schouest L., Fletcher W. H. Morphologic analysis of long-term bone marrow cultures that support B-lymphopoiesis or myelopoiesis. Cell Tissue Res. 1985;239(2):375–382. doi: 10.1007/BF00218018. [DOI] [PubMed] [Google Scholar]
  10. Gillis S., Smith K. A. Long term culture of tumour-specific cytotoxic T cells. Nature. 1977 Jul 14;268(5616):154–156. doi: 10.1038/268154a0. [DOI] [PubMed] [Google Scholar]
  11. Hayakawa K., Hardy R. R., Herzenberg L. A., Herzenberg L. A. Progenitors for Ly-1 B cells are distinct from progenitors for other B cells. J Exp Med. 1985 Jun 1;161(6):1554–1568. doi: 10.1084/jem.161.6.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hayakawa K., Hardy R. R., Parks D. R., Herzenberg L. A. The "Ly-1 B" cell subpopulation in normal immunodefective, and autoimmune mice. J Exp Med. 1983 Jan 1;157(1):202–218. doi: 10.1084/jem.157.1.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herzenberg L. A., Stall A. M., Lalor P. A., Sidman C., Moore W. A., Parks D. R., Herzenberg L. A. The Ly-1 B cell lineage. Immunol Rev. 1986 Oct;93:81–102. doi: 10.1111/j.1600-065x.1986.tb01503.x. [DOI] [PubMed] [Google Scholar]
  14. Hunt P., Robertson D., Weiss D., Rennick D., Lee F., Witte O. N. A single bone marrow-derived stromal cell type supports the in vitro growth of early lymphoid and myeloid cells. Cell. 1987 Mar 27;48(6):997–1007. doi: 10.1016/0092-8674(87)90708-2. [DOI] [PubMed] [Google Scholar]
  15. Koyasu S., Yodoi J., Nikaido T., Tagaya Y., Taniguchi Y., Honjo T., Yahara I. Expression of interleukin 2 receptors on interleukin 3-dependent cell lines. J Immunol. 1986 Feb 1;136(3):984–987. [PubMed] [Google Scholar]
  16. Le Gros G. S., Gillis S., Watson J. D. Induction of IL 2 responsiveness in a murine IL 3-dependent cell line. J Immunol. 1985 Dec;135(6):4009–4014. [PubMed] [Google Scholar]
  17. Manohar V., Brown E., Leiserson W. M., Chused T. M. Expression of Lyt-1 by a subset of B lymphocytes. J Immunol. 1982 Aug;129(2):532–538. [PubMed] [Google Scholar]
  18. McKearn J. P., McCubrey J., Fagg B. Enrichment of hematopoietic precursor cells and cloning of multipotential B-lymphocyte precursors. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7414–7418. doi: 10.1073/pnas.82.21.7414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Muller-Sieburg C. E., Whitlock C. A., Weissman I. L. Isolation of two early B lymphocyte progenitors from mouse marrow: a committed pre-pre-B cell and a clonogenic Thy-1-lo hematopoietic stem cell. Cell. 1986 Feb 28;44(4):653–662. doi: 10.1016/0092-8674(86)90274-6. [DOI] [PubMed] [Google Scholar]
  20. Nishikawa S., Sasaki Y., Kina T., Amagai T., Katsura Y. A monoclonal antibody against Igh6-4 determinant. Immunogenetics. 1986;23(2):137–139. doi: 10.1007/BF00377976. [DOI] [PubMed] [Google Scholar]
  21. Ozato K., Mayer N., Sachs D. H. Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J Immunol. 1980 Feb;124(2):533–540. [PubMed] [Google Scholar]
  22. Paige C. J. Surface immunoglobulin-negative B-cell precursors detected by formation of antibody-secreting colonies in agar. Nature. 1983 Apr 21;302(5910):711–713. doi: 10.1038/302711a0. [DOI] [PubMed] [Google Scholar]
  23. Palacios R., Kiefer M., Brockhaus M., Karjalainen K., Dembić Z., Kisielow P., von Boehmer H. Molecular, cellular, and functional properties of bone marrow T lymphocyte progenitor clones. J Exp Med. 1987 Jul 1;166(1):12–32. doi: 10.1084/jem.166.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Palacios R., Leu T. Both cloned interleukin 2 and purified interleukin 1 are required for optimal growth of purified L3T4+ and Lyt 2+ lymphocytes initiated by concanavalin A. Cell Immunol. 1985 Sep;94(2):369–382. doi: 10.1016/0008-8749(85)90261-8. [DOI] [PubMed] [Google Scholar]
  25. Palacios R., Leu T. CC11: a monoclonal antibody specific for interleukin 3-sensitive mouse cells defines two major populations of B cell precursors in the bone marrow. Immunol Rev. 1986 Oct;93:125–146. doi: 10.1111/j.1600-065x.1986.tb01505.x. [DOI] [PubMed] [Google Scholar]
  26. Palacios R., Neri T., Brockhaus M. Monoclonal antibodies specific for interleukin 3-sensitive murine cells. J Exp Med. 1986 Feb 1;163(2):369–382. doi: 10.1084/jem.163.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Palacios R., Sideras P., von Boehmer H. Recombinant interleukin 4/BSF-1 promotes growth and differentiation of intrathymic T cell precursors from fetal mice in vitro. EMBO J. 1987 Jan;6(1):91–95. doi: 10.1002/j.1460-2075.1987.tb04723.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Palacios R., Steinmetz M. Il-3-dependent mouse clones that express B-220 surface antigen, contain Ig genes in germ-line configuration, and generate B lymphocytes in vivo. Cell. 1985 Jul;41(3):727–734. doi: 10.1016/s0092-8674(85)80053-2. [DOI] [PubMed] [Google Scholar]
  29. Raff M. C., Megson M., Owen J. J., Cooper M. D. Early production of intracellular IgM by B-lymphocyte precursors in mouse. Nature. 1976 Jan 22;259(5540):224–226. doi: 10.1038/259224a0. [DOI] [PubMed] [Google Scholar]
  30. Raff M. C., Sternberg M., Taylor R. B. Immunoglobulin determinants on the surface of mouse lymphoid cells. Nature. 1970 Feb 7;225(5232):553–554. doi: 10.1038/225553a0. [DOI] [PubMed] [Google Scholar]
  31. Robb R. J., Greene W. C., Rusk C. M. Low and high affinity cellular receptors for interleukin 2. Implications for the level of Tac antigen. J Exp Med. 1984 Oct 1;160(4):1126–1146. doi: 10.1084/jem.160.4.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rolink A. G., Radaszkiewicz T., Melchers F. The autoantigen-binding B cell repertoires of normal and of chronically graft-versus-host-diseased mice. J Exp Med. 1987 Jun 1;165(6):1675–1687. doi: 10.1084/jem.165.6.1675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Sideras P., Palacios R. Bone marrow pro-T and pro-B lymphocyte clones express functional receptors for interleukin (IL) 3 and IL 4/BSF-1 and nonfunctional receptors for IL 2. Eur J Immunol. 1987 Feb;17(2):217–221. doi: 10.1002/eji.1830170211. [DOI] [PubMed] [Google Scholar]
  35. Spalding D. M., Griffin J. A. Different pathways of differentiation of pre-B cell lines are induced by dendritic cells and T cells from different lymphoid tissues. Cell. 1986 Feb 14;44(3):507–515. doi: 10.1016/0092-8674(86)90472-1. [DOI] [PubMed] [Google Scholar]
  36. Springer T., Galfré G., Secher D. S., Milstein C. Mac-1: a macrophage differentiation antigen identified by monoclonal antibody. Eur J Immunol. 1979 Apr;9(4):301–306. doi: 10.1002/eji.1830090410. [DOI] [PubMed] [Google Scholar]
  37. Steinmetz M., Zachau H. G. Two rearranged immunoglobulin kappa light chain genes in one mouse myeloma. Nucleic Acids Res. 1980 Apr 25;8(8):1693–1707. doi: 10.1093/nar/8.8.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Storb U., Arp B. Methylation patterns of immunoglobulin genes in lymphoid cells: correlation of expression and differentiation with undermethylation. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6642–6646. doi: 10.1073/pnas.80.21.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Takemori T., Rajewsky K. Lambda chain expression at different stages of ontogeny in C57BL/6, BALB/c and SJL mice. Eur J Immunol. 1981 Aug;11(8):618–625. doi: 10.1002/eji.1830110806. [DOI] [PubMed] [Google Scholar]
  40. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  41. Whitlock C. A., Tidmarsh G. F., Muller-Sieburg C., Weissman I. L. Bone marrow stromal cell lines with lymphopoietic activity express high levels of a pre-B neoplasia-associated molecule. Cell. 1987 Mar 27;48(6):1009–1021. doi: 10.1016/0092-8674(87)90709-4. [DOI] [PubMed] [Google Scholar]
  42. Whitlock C. A., Witte O. N. Long-term culture of B lymphocytes and their precursors from murine bone marrow. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3608–3612. doi: 10.1073/pnas.79.11.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yancopoulos G. D., Blackwell T. K., Suh H., Hood L., Alt F. W. Introduced T cell receptor variable region gene segments recombine in pre-B cells: evidence that B and T cells use a common recombinase. Cell. 1986 Jan 31;44(2):251–259. doi: 10.1016/0092-8674(86)90759-2. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES