Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Aug 1;134(3):771–782. doi: 10.1083/jcb.134.3.771

Identification of stromal cell products that interact with pre-B cells

PMCID: PMC2120935  PMID: 8707854

Abstract

Our understanding of lympho-hematopoietic microenvironments is incomplete, and a new cloning strategy was developed to identify molecules that bind to B lineage lymphocyte precursors. A cell sorting procedure was used for initial enrichment of cDNAs from stromal cell mRNA that contained signal sequences and were therefore likely to encode transmembrane or secreted proteins. A second step involved expression of the library as soluble Ig fusion proteins. Finally, pools representing these proteins were screened for the ability to recognize pre-B cells. This approach resulted in the cloning of biglycan, syndecan 4, collagen type I, clusterin, matrix glycoprotein sc1, osteonectin, and one unknown molecule (designated SIM). The full-length cDNA of SIM revealed that it is a type I transmembrane protein, and its intracellular domain has weak homology with myosin heavy chain and related proteins. Staining of established cell lines and freshly isolated hematopoietic cells with the Ig fusion proteins revealed distinct patterns of reactivity and differential dependence on divalent cations. Biglycan-, sc1-, and SIM-Ig fusion proteins selectively increased interleukin 7-dependent proliferation of pre-B cells. Overexpression of the entire SIM protein affected the morphology of 293T cells, while expression of just the extracellular portion was without effect. Thus, a series of stromal cell surface molecules has been identified that interact with blood cell precursors. Three of them promoted the survival and/or proliferation of pre-B cells in culture, and all merit further study in relation to lympho-hematopoiesis.

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. Aruffo A., Stamenkovic I., Melnick M., Underhill C. B., Seed B. CD44 is the principal cell surface receptor for hyaluronate. Cell. 1990 Jun 29;61(7):1303–1313. doi: 10.1016/0092-8674(90)90694-a. [DOI] [PubMed] [Google Scholar]
  2. Brunner G., Metz C. N., Nguyen H., Gabrilove J., Patel S. R., Davitz M. A., Rifkin D. B., Wilson E. L. An endogenous glycosylphosphatidylinositol-specific phospholipase D releases basic fibroblast growth factor-heparan sulfate proteoglycan complexes from human bone marrow cultures. Blood. 1994 Apr 15;83(8):2115–2125. [PubMed] [Google Scholar]
  3. Cardin A. D., Weintraub H. J. Molecular modeling of protein-glycosaminoglycan interactions. Arteriosclerosis. 1989 Jan-Feb;9(1):21–32. doi: 10.1161/01.atv.9.1.21. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. DeLisser H. M., Yan H. C., Newman P. J., Muller W. A., Buck C. A., Albelda S. M. Platelet/endothelial cell adhesion molecule-1 (CD31)-mediated cellular aggregation involves cell surface glycosaminoglycans. J Biol Chem. 1993 Jul 25;268(21):16037–16046. [PubMed] [Google Scholar]
  6. Eisenberg D., Schwarz E., Komaromy M., Wall R. Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol. 1984 Oct 15;179(1):125–142. doi: 10.1016/0022-2836(84)90309-7. [DOI] [PubMed] [Google Scholar]
  7. French L. E., Chonn A., Ducrest D., Baumann B., Belin D., Wohlwend A., Kiss J. Z., Sappino A. P., Tschopp J., Schifferli J. A. Murine clusterin: molecular cloning and mRNA localization of a gene associated with epithelial differentiation processes during embryogenesis. J Cell Biol. 1993 Sep;122(5):1119–1130. doi: 10.1083/jcb.122.5.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. French L. E., Sappino A. P., Tschopp J., Schifferli J. A. Clusterin gene expression in the rat thymus is not modulated by dexamethasone treatment. Immunology. 1994 Jun;82(2):328–331. [PMC free article] [PubMed] [Google Scholar]
  9. Hildebrand A., Romarís M., Rasmussen L. M., Heinegård D., Twardzik D. R., Border W. A., Ruoslahti E. Interaction of the small interstitial proteoglycans biglycan, decorin and fibromodulin with transforming growth factor beta. Biochem J. 1994 Sep 1;302(Pt 2):527–534. doi: 10.1042/bj3020527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  11. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  12. Johnston I. G., Paladino T., Gurd J. W., Brown I. R. Molecular cloning of SC1: a putative brain extracellular matrix glycoprotein showing partial similarity to osteonectin/BM40/SPARC. Neuron. 1990 Jan;4(1):165–176. doi: 10.1016/0896-6273(90)90452-l. [DOI] [PubMed] [Google Scholar]
  13. Kamo I., Kikuchi A., Nonaka I., Yamada E., Kondo J. Haemopoietic activity associated with biglycan like proteoglycan. Biochem Biophys Res Commun. 1993 Sep 15;195(2):1119–1126. doi: 10.1006/bbrc.1993.2160. [DOI] [PubMed] [Google Scholar]
  14. Katoh S., Zheng Z., Oritani K., Shimozato T., Kincade P. W. Glycosylation of CD44 negatively regulates its recognition of hyaluronan. J Exp Med. 1995 Aug 1;182(2):419–429. doi: 10.1084/jem.182.2.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kim C. W., Goldberger O. A., Gallo R. L., Bernfield M. Members of the syndecan family of heparan sulfate proteoglycans are expressed in distinct cell-, tissue-, and development-specific patterns. Mol Biol Cell. 1994 Jul;5(7):797–805. doi: 10.1091/mbc.5.7.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kimura K., Matsubara H., Sogoh S., Kita Y., Sakata T., Nishitani Y., Watanabe S., Hamaoka T., Fujiwara H. Role of glycosaminoglycans in the regulation of T cell proliferation induced by thymic stroma-derived T cell growth factor. J Immunol. 1991 Apr 15;146(8):2618–2624. [PubMed] [Google Scholar]
  17. Kincade P. W. Molecular interactions between stromal cells and B lymphocyte precursors. Semin Immunol. 1991 Nov;3(6):379–390. [PubMed] [Google Scholar]
  18. Kitamura D., Kudo A., Schaal S., Müller W., Melchers F., Rajewsky K. A critical role of lambda 5 protein in B cell development. Cell. 1992 May 29;69(5):823–831. doi: 10.1016/0092-8674(92)90293-l. [DOI] [PubMed] [Google Scholar]
  19. Knudson W., Bartnik E., Knudson C. B. Assembly of pericellular matrices by COS-7 cells transfected with CD44 lymphocyte-homing receptor genes. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4003–4007. doi: 10.1073/pnas.90.9.4003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koenigsmann M., Griffin J. D., DiCarlo J., Cannistra S. A. Myeloid and erythroid progenitor cells from normal bone marrow adhere to collagen type I. Blood. 1992 Feb 1;79(3):657–665. [PubMed] [Google Scholar]
  21. Kojima T., Shworak N. W., Rosenberg R. D. Molecular cloning and expression of two distinct cDNA-encoding heparan sulfate proteoglycan core proteins from a rat endothelial cell line. J Biol Chem. 1992 Mar 5;267(7):4870–4877. [PubMed] [Google Scholar]
  22. Lane T. F., Sage E. H. The biology of SPARC, a protein that modulates cell-matrix interactions. FASEB J. 1994 Feb;8(2):163–173. [PubMed] [Google Scholar]
  23. Lee G., Namen A. E., Gillis S., Ellingsworth L. R., Kincade P. W. Normal B cell precursors responsive to recombinant murine IL-7 and inhibition of IL-7 activity by transforming growth factor-beta. J Immunol. 1989 Jun 1;142(11):3875–3883. [PubMed] [Google Scholar]
  24. Lesley J., He Q., Miyake K., Hamann A., Hyman R., Kincade P. W. Requirements for hyaluronic acid binding by CD44: a role for the cytoplasmic domain and activation by antibody. J Exp Med. 1992 Jan 1;175(1):257–266. doi: 10.1084/jem.175.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lesley J., Hyman R., Kincade P. W. CD44 and its interaction with extracellular matrix. Adv Immunol. 1993;54:271–335. doi: 10.1016/s0065-2776(08)60537-4. [DOI] [PubMed] [Google Scholar]
  26. Long M. W. Blood cell cytoadhesion molecules. Exp Hematol. 1992 Mar;20(3):288–301. [PubMed] [Google Scholar]
  27. Long M. W., Williams J. L., Mann K. G. Expression of human bone-related proteins in the hematopoietic microenvironment. J Clin Invest. 1990 Nov;86(5):1387–1395. doi: 10.1172/JCI114852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lortat-Jacob H., Grimaud J. A. Interferon-gamma C-terminal function: new working hypothesis. Heparan sulfate and heparin, new targets for IFN-gamma, protect, relax the cytokine and regulate its activity. Cell Mol Biol. 1991;37(3):253–260. [PubMed] [Google Scholar]
  29. Luikart S. D., Kenney P. M., Oegema T. R. Human bone marrow heparan sulfate induces leukemia cell differentiation. Connect Tissue Res. 1995;31(2):99–107. doi: 10.3109/03008209509028397. [DOI] [PubMed] [Google Scholar]
  30. Lund J., Winter G., Jones P. T., Pound J. D., Tanaka T., Walker M. R., Artymiuk P. J., Arata Y., Burton D. R., Jefferis R. Human Fc gamma RI and Fc gamma RII interact with distinct but overlapping sites on human IgG. J Immunol. 1991 Oct 15;147(8):2657–2662. [PubMed] [Google Scholar]
  31. Mason I. J., Taylor A., Williams J. G., Sage H., Hogan B. L. Evidence from molecular cloning that SPARC, a major product of mouse embryo parietal endoderm, is related to an endothelial cell 'culture shock' glycoprotein of Mr 43,000. EMBO J. 1986 Jul;5(7):1465–1472. doi: 10.1002/j.1460-2075.1986.tb04383.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Masumoto A., Hemler M. E. Multiple activation states of VLA-4. Mechanistic differences between adhesion to CS1/fibronectin and to vascular cell adhesion molecule-1. J Biol Chem. 1993 Jan 5;268(1):228–234. [PubMed] [Google Scholar]
  33. Medina K. L., Smithson G., Kincade P. W. Suppression of B lymphopoiesis during normal pregnancy. J Exp Med. 1993 Nov 1;178(5):1507–1515. doi: 10.1084/jem.178.5.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Melchers F., Haasner D., Grawunder U., Kalberer C., Karasuyama H., Winkler T., Rolink A. G. Roles of IgH and L chains and of surrogate H and L chains in the development of cells of the B lymphocyte lineage. Annu Rev Immunol. 1994;12:209–225. doi: 10.1146/annurev.iy.12.040194.001233. [DOI] [PubMed] [Google Scholar]
  35. Miyake K., Medina K. L., Hayashi S., Ono S., Hamaoka T., Kincade P. W. Monoclonal antibodies to Pgp-1/CD44 block lympho-hemopoiesis in long-term bone marrow cultures. J Exp Med. 1990 Feb 1;171(2):477–488. doi: 10.1084/jem.171.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Miyake K., Medina K., Ishihara K., Kimoto M., Auerbach R., Kincade P. W. A VCAM-like adhesion molecule on murine bone marrow stromal cells mediates binding of lymphocyte precursors in culture. J Cell Biol. 1991 Aug;114(3):557–565. doi: 10.1083/jcb.114.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Miyake K., Weissman I. L., Greenberger J. S., Kincade P. W. Evidence for a role of the integrin VLA-4 in lympho-hemopoiesis. J Exp Med. 1991 Mar 1;173(3):599–607. doi: 10.1084/jem.173.3.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Mizushima S., Nagata S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 1990 Sep 11;18(17):5322–5322. doi: 10.1093/nar/18.17.5322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Nishiguchi S., Joh T., Horie K., Zou Z., Yasunaga T., Shimada K. A survey of genes expressed in undifferentiated mouse embryonal carcinoma F9 cells: characterization of low-abundance mRNAs. J Biochem. 1994 Jul;116(1):128–139. doi: 10.1093/oxfordjournals.jbchem.a124485. [DOI] [PubMed] [Google Scholar]
  40. Phillips C. L., Lever L. W., Pinnell S. R., Quarles L. D., Wenstrup R. J. Construction of a full-length murine pro alpha 2(I) collagen cDNA by the polymerase chain reaction. J Invest Dermatol. 1991 Dec;97(6):980–984. doi: 10.1111/1523-1747.ep12491894. [DOI] [PubMed] [Google Scholar]
  41. Rapraeger A. C., Krufka A., Olwin B. B. Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science. 1991 Jun 21;252(5013):1705–1708. doi: 10.1126/science.1646484. [DOI] [PubMed] [Google Scholar]
  42. Rau W., Just W., Vetter U., Vogel W. A dinucleotide repeat in the mouse biglycan gene (EST) on the X chromosome. Mamm Genome. 1994 Jun;5(6):395–396. doi: 10.1007/BF00356564. [DOI] [PubMed] [Google Scholar]
  43. Rezaie A. R., Fiore M. M., Neuenschwander P. F., Esmon C. T., Morrissey J. H. Expression and purification of a soluble tissue factor fusion protein with an epitope for an unusual calcium-dependent antibody. Protein Expr Purif. 1992 Dec;3(6):453–460. doi: 10.1016/1046-5928(92)90062-2. [DOI] [PubMed] [Google Scholar]
  44. Roberts R., Gallagher J., Spooncer E., Allen T. D., Bloomfield F., Dexter T. M. Heparan sulphate bound growth factors: a mechanism for stromal cell mediated haemopoiesis. Nature. 1988 Mar 24;332(6162):376–378. doi: 10.1038/332376a0. [DOI] [PubMed] [Google Scholar]
  45. Rosenberg M. E., Dvergsten J., Correa-Rotter R. Clusterin: an enigmatic protein recruited by diverse stimuli. J Lab Clin Med. 1993 Feb;121(2):205–214. [PubMed] [Google Scholar]
  46. Sanderson R. D., Lalor P., Bernfield M. B lymphocytes express and lose syndecan at specific stages of differentiation. Cell Regul. 1989 Nov;1(1):27–35. doi: 10.1091/mbc.1.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sanderson R. D., Sneed T. B., Young L. A., Sullivan G. L., Lander A. D. Adhesion of B lymphoid (MPC-11) cells to type I collagen is mediated by integral membrane proteoglycan, syndecan. J Immunol. 1992 Jun 15;148(12):3902–3911. [PubMed] [Google Scholar]
  48. Skarnes W. C., Moss J. E., Hurtley S. M., Beddington R. S. Capturing genes encoding membrane and secreted proteins important for mouse development. Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6592–6596. doi: 10.1073/pnas.92.14.6592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Smithson G., Medina K., Ponting I., Kincade P. W. Estrogen suppresses stromal cell-dependent lymphopoiesis in culture. J Immunol. 1995 Oct 1;155(7):3409–3417. [PubMed] [Google Scholar]
  50. Stanley M. J., Liebersbach B. F., Liu W., Anhalt D. J., Sanderson R. D. Heparan sulfate-mediated cell aggregation. Syndecans-1 and -4 mediate intercellular adhesion following their transfection into human B lymphoid cells. J Biol Chem. 1995 Mar 10;270(10):5077–5083. doi: 10.1074/jbc.270.10.5077. [DOI] [PubMed] [Google Scholar]
  51. Sugahara H., Kanakura Y., Furitsu T., Ishihara K., Oritani K., Ikeda H., Kitayama H., Ishikawa J., Hashimoto K., Kanayama Y. Induction of programmed cell death in human hematopoietic cell lines by fibronectin via its interaction with very late antigen 5. J Exp Med. 1994 Jun 1;179(6):1757–1766. doi: 10.1084/jem.179.6.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tashiro K., Tada H., Heilker R., Shirozu M., Nakano T., Honjo T. Signal sequence trap: a cloning strategy for secreted proteins and type I membrane proteins. Science. 1993 Jul 30;261(5121):600–603. doi: 10.1126/science.8342023. [DOI] [PubMed] [Google Scholar]
  53. Tsuda M., Sakiyama T., Endo H., Kitagawa T. The primary structure of mouse saposin. Biochem Biophys Res Commun. 1992 May 15;184(3):1266–1272. doi: 10.1016/s0006-291x(05)80019-1. [DOI] [PubMed] [Google Scholar]
  54. Unkeless J. C. Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors. J Exp Med. 1979 Sep 19;150(3):580–596. doi: 10.1084/jem.150.3.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Verfaillie C., Hurley R., Bhatia R., McCarthy J. B. Role of bone marrow matrix in normal and abnormal hematopoiesis. Crit Rev Oncol Hematol. 1994 Jun;16(3):201–224. doi: 10.1016/1040-8428(94)90071-x. [DOI] [PubMed] [Google Scholar]
  56. Wilson M. R., Easterbrook-Smith S. B. Clusterin binds by a multivalent mechanism to the Fc and Fab regions of IgG. Biochim Biophys Acta. 1992 Oct 20;1159(3):319–326. doi: 10.1016/0167-4838(92)90062-i. [DOI] [PubMed] [Google Scholar]
  57. Zuckerman K. S., Rhodes R. K., Goodrum D. D., Patel V. R., Sparks B., Wells J., Wicha M. S., Mayo L. A. Inhibition of collagen deposition in the extracellular matrix prevents the establishment of a stroma supportive of hematopoiesis in long-term murine bone marrow cultures. J Clin Invest. 1985 Mar;75(3):970–975. doi: 10.1172/JCI111798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES