Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1989 Dec 1;170(6):1973–1986. doi: 10.1084/jem.170.6.1973

One synchronous wave of B cell development in mouse fetal liver changes at day 16 of gestation from dependence to independence of a stromal cell environment

PMCID: PMC2189533  PMID: 2584932

Abstract

Precursor cells of the B lineage can be enriched from mouse fetal liver by FACS with the aid of the pre-B cell-specific mAb G-5-2. The cells are concomitantly enriched for cells expressing the pre-B cell-specific gene lambda 5, and for cells developing to LPS-reactive mature B cells. The enriched purified precursors are not influenced by rIL-2 through - 7, alone or in combination, to develop to mitogen-reactive, sIg+ cells. Marginal proliferation of the precursors is observed in response to IL- 3 plus -4, and IL-6 plus -7, and this does not change in the presence of stromal cells. Development to mitogen-reactive, sIg+ cells is dependent on interactions with embryonic stromal cells from fetal liver. Two mAbs raised against the stromal cells inhibit this development. Two phases of precursor cell development can be distinguished in fetal liver. Between days 13 and 15 of gestation, it is dependent on stromal cell interactions, thereafter, from days 16 to 19, it is independent. A sudden increase in the number of mitogen- reactive, sIg+ B lineage cells occurs within 24 h between days 16 and 17. All these results indicate that B cell development occurs in one wave with synchronous steps of changes from a mitogen-insensitive, sIg- , stromal cell dependent to a mitogen-reactive, sIg+, stromal cell- independent B lineage line.

Full Text

The Full Text of this article is available as a PDF (978.4 KB).

Selected References

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

  1. Alt F., Rosenberg N., Lewis S., Thomas E., Baltimore D. Organization and reorganization of immunoglobulin genes in A-MULV-transformed cells: rearrangement of heavy but not light chain genes. Cell. 1981 Dec;27(2 Pt 1):381–390. doi: 10.1016/0092-8674(81)90421-9. [DOI] [PubMed] [Google Scholar]
  2. Björck L., Kronvall G. Purification and some properties of streptococcal protein G, a novel IgG-binding reagent. J Immunol. 1984 Aug;133(2):969–974. [PubMed] [Google Scholar]
  3. Clevers H., Alarcon B., Wileman T., Terhorst C. The T cell receptor/CD3 complex: a dynamic protein ensemble. Annu Rev Immunol. 1988;6:629–662. doi: 10.1146/annurev.iy.06.040188.003213. [DOI] [PubMed] [Google Scholar]
  4. Dorshkind K., Johnson A., Collins L., Keller G. M., Phillips R. A. Generation of purified stromal cell cultures that support lymphoid and myeloid precursors. J Immunol Methods. 1986 May 1;89(1):37–47. doi: 10.1016/0022-1759(86)90029-3. [DOI] [PubMed] [Google Scholar]
  5. Gisler R. H., Holländer G., Söderberg A. Functional maturation of murine B lymphocyte precursors. I. Selection of adherent cell-dependent precursors from bone marrow and fetal liver. J Immunol. 1987 Apr 15;138(8):2427–2432. [PubMed] [Google Scholar]
  6. Gisler R. H., Schlienger C., Söderberg A., Ledermann F., Lambris J. D. Functional maturation of murine B lymphocyte precursors--III. Soluble factors involved in the regulation of growth and differentiation. Mol Immunol. 1988 Nov;25(11):1113–1127. doi: 10.1016/0161-5890(88)90146-0. [DOI] [PubMed] [Google Scholar]
  7. Gisler R. H., Söderberg A., Kamber M. Functional maturation of murine B lymphocyte precursors. II. Analysis of cells required from the bone marrow microenvironment. J Immunol. 1987 Apr 15;138(8):2433–2438. [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. JERNE N. K., NORDIN A. A. Plaque formation in agar by single antibody-producing cells. Science. 1963 Apr 26;140(3565):405–405. [PubMed] [Google Scholar]
  11. Karasuyama H., Melchers F. Establishment of mouse cell lines which constitutively secrete large quantities of interleukin 2, 3, 4 or 5, using modified cDNA expression vectors. Eur J Immunol. 1988 Jan;18(1):97–104. doi: 10.1002/eji.1830180115. [DOI] [PubMed] [Google Scholar]
  12. Kinashi T., Inaba K., Tsubata T., Tashiro K., Palacios R., Honjo T. Differentiation of an interleukin 3-dependent precursor B-cell clone into immunoglobulin-producing cells in vitro. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4473–4477. doi: 10.1073/pnas.85.12.4473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kincade P. W., Lee G., Paige C. J., Scheid M. P. Cellular interactions affecting the maturation of murine B lymphocyte precursors in vitro. J Immunol. 1981 Jul;127(1):255–260. [PubMed] [Google Scholar]
  14. 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]
  15. Maki R., Kearney J., Paige C., Tonegawa S. Immunoglobulin gene rearrangement in immature B cells. Science. 1980 Sep 19;209(4463):1366–1369. doi: 10.1126/science.6774416. [DOI] [PubMed] [Google Scholar]
  16. Melchers F. B lymphocyte development in fetal liver. I. Development of reactivities to B cell mitogens "in vivo" and "in vitro". Eur J Immunol. 1977 Jul;7(7):476–481. doi: 10.1002/eji.1830070714. [DOI] [PubMed] [Google Scholar]
  17. Melchers F. B lymphocyte development in fetal liver. II. Frequencies of precursor B cells during gestation. Eur J Immunol. 1977 Jul;7(7):482–486. doi: 10.1002/eji.1830070715. [DOI] [PubMed] [Google Scholar]
  18. Melchers F., Von Boehmer H., Phillips R. A. B-lymphocyte subpopulations in the mouse. Organ distribution and ontogeny of immunoglobulin-synthesizing and of mitogen-sensitive cells. Transplant Rev. 1975;25:26–58. doi: 10.1111/j.1600-065x.1975.tb00725.x. [DOI] [PubMed] [Google Scholar]
  19. Namen A. E., Lupton S., Hjerrild K., Wignall J., Mochizuki D. Y., Schmierer A., Mosley B., March C. J., Urdal D., Gillis S. Stimulation of B-cell progenitors by cloned murine interleukin-7. Nature. 1988 Jun 9;333(6173):571–573. doi: 10.1038/333571a0. [DOI] [PubMed] [Google Scholar]
  20. Namen A. E., Schmierer A. E., March C. J., Overell R. W., Park L. S., Urdal D. L., Mochizuki D. Y. B cell precursor growth-promoting activity. Purification and characterization of a growth factor active on lymphocyte precursors. J Exp Med. 1988 Mar 1;167(3):988–1002. doi: 10.1084/jem.167.3.988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Owen J. J., Cooper M. D., Raff M. C. In vitro generation of B lymphocytes in mouse foetal liver, a mammalian 'bursa equivalent'. Nature. 1974 May 24;249(455):361–363. doi: 10.1038/249361a0. [DOI] [PubMed] [Google Scholar]
  22. Paige C. J., Gisler R. H., McKearn J. P., Iscove N. N. Differentiation of murine B cell precursors in agar culture. Frequency, surface marker analysis and requirements for growth of clonable pre-B cells. Eur J Immunol. 1984 Nov;14(11):979–987. doi: 10.1002/eji.1830141104. [DOI] [PubMed] [Google Scholar]
  23. Palacios R., Henson G., Steinmetz M., McKearn J. P. Interleukin-3 supports growth of mouse pre-B-cell clones in vitro. Nature. 1984 May 10;309(5964):126–131. doi: 10.1038/309126a0. [DOI] [PubMed] [Google Scholar]
  24. Palacios R., Karasuyama H., Rolink A. Ly1+ PRO-B lymphocyte clones. Phenotype, growth requirements and differentiation in vitro and in vivo. EMBO J. 1987 Dec 1;6(12):3687–3693. doi: 10.1002/j.1460-2075.1987.tb02702.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Palacios R., Stuber S., Rolink A. The epigenetic influences of bone marrow and fetal liver stroma cells on the developmental potential of Ly-1+ pro-B lymphocyte clones. Eur J Immunol. 1989 Feb;19(2):347–356. doi: 10.1002/eji.1830190220. [DOI] [PubMed] [Google Scholar]
  27. Park Y. H., Osmond D. G. Phenotype and proliferation of early B lymphocyte precursor cells in mouse bone marrow. J Exp Med. 1987 Feb 1;165(2):444–458. doi: 10.1084/jem.165.2.444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pietrangeli C. E., Hayashi S., Kincade P. W. Stromal cell lines which support lymphocyte growth: characterization, sensitivity to radiation and responsiveness to growth factors. Eur J Immunol. 1988 Jun;18(6):863–872. doi: 10.1002/eji.1830180606. [DOI] [PubMed] [Google Scholar]
  29. Reynolds J. D., Morris B. The evolution and involution of Peyer's patches in fetal and postnatal sheep. Eur J Immunol. 1983 Aug;13(8):627–635. doi: 10.1002/eji.1830130805. [DOI] [PubMed] [Google Scholar]
  30. Rolink A. G., Melchers F., Palacios R. Monoclonal antibodies reactive with the mouse interleukin 5 receptor. J Exp Med. 1989 May 1;169(5):1693–1701. doi: 10.1084/jem.169.5.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sakaguchi N., Berger C. N., Melchers F. Isolation of a cDNA copy of an RNA species expressed in murine pre-B cells. EMBO J. 1986 Sep;5(9):2139–2147. doi: 10.1002/j.1460-2075.1986.tb04477.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sakaguchi N., Kashiwamura S., Kimoto M., Thalmann P., Melchers F. B lymphocyte lineage-restricted expression of mb-1, a gene with CD3-like structural properties. EMBO J. 1988 Nov;7(11):3457–3464. doi: 10.1002/j.1460-2075.1988.tb03220.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sakaguchi N., Melchers F. Lambda 5, a new light-chain-related locus selectively expressed in pre-B lymphocytes. Nature. 1986 Dec 11;324(6097):579–582. doi: 10.1038/324579a0. [DOI] [PubMed] [Google Scholar]
  34. Sibley C. H., Ewald S. J., Kehry M. R., Douglas R. H., Raschke W. C., Hood L. E. Characterization of multiple immunoglobulin mu-chains synthesized by two clones of a B cell lymphoma. J Immunol. 1980 Nov;125(5):2097–2105. [PubMed] [Google Scholar]
  35. Strasser A. PB76: a novel surface glycoprotein preferentially expressed on mouse pre-B cells and plasma cells detected by the monoclonal antibody G-5-2. Eur J Immunol. 1988 Nov;18(11):1803–1810. doi: 10.1002/eji.1830181123. [DOI] [PubMed] [Google Scholar]
  36. Takeda S., Gillis S., Palacios R. In vitro effects of recombinant interleukin 7 on growth and differentiation of bone marrow pro-B- and pro-T-lymphocyte clones and fetal thymocyte clones. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1634–1638. doi: 10.1073/pnas.86.5.1634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Warner N. L., Daley M. J., Richey J., Spellman C. Flow cytometry analysis of murine B cell lymphoma differentiation. Immunol Rev. 1979;48:197–243. doi: 10.1111/j.1600-065x.1979.tb00304.x. [DOI] [PubMed] [Google Scholar]
  38. Whitlock C., Denis K., Robertson D., Witte O. In vitro analysis of murine B-cell development. Annu Rev Immunol. 1985;3:213–235. doi: 10.1146/annurev.iy.03.040185.001241. [DOI] [PubMed] [Google Scholar]
  39. Witte P. L., Robinson M., Henley A., Low M. G., Stiers D. L., Perkins S., Fleischman R. A., Kincade P. W. Relationships between B-lineage lymphocytes and stromal cells in long-term bone marrow cultures. Eur J Immunol. 1987 Oct;17(10):1473–1484. doi: 10.1002/eji.1830171014. [DOI] [PubMed] [Google Scholar]

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

RESOURCES