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. 1990 Feb 1;171(2):477–488. doi: 10.1084/jem.171.2.477

Monoclonal antibodies to Pgp-1/CD44 block lympho-hemopoiesis in long- term bone marrow cultures

PMCID: PMC2187725  PMID: 2406365

Abstract

A new panel of mAbs was prepared to a stromal cell line known to support lymphocytes in Whitlock-Witte type long-term bone marrow cultures. These antibodies were then screened with a cell adhesion assay and four were selected that inhibited the binding of B lineage cells to stromal cell monolayers. Immunofluorescent and biochemical analyses revealed that these new antibodies detected epitopes of the previously described Pgp-1/CD44 antigen complex. Addition of Pgp-1/CD44 antibodies to Dexter-type long-term bone marrow cultures completely prevented emergence of myeloid cells and they also blocked lymphocyte growth in Whitlock-Witte type cultures. mAbs MEL-14, LFA-1, and CD45R did not inhibit under the same conditions and there was no apparent relationship to Ig isotype. Adherent layers in treated cultures were not unusual in terms of morphology and the antibodies did not affect factor-dependent replication of lymphoid or myeloid progenitor cells. Therefore, the mechanism of inhibition may not involve direct toxicity to precursors or microenvironmental elements. Previous studies in humans and mice have implicated Pgp-1/CD44-related glycoproteins in the migration of peripheral lymphoid cells, as well as interactions of cells with the extracellular matrix. These findings suggest that they may also be critical for formation of lymphoid and myeloid cells within bone marrow.

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Selected References

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

  1. Budd R. C., Cerottini J. C., Horvath C., Bron C., Pedrazzini T., Howe R. C., MacDonald H. R. Distinction of virgin and memory T lymphocytes. Stable acquisition of the Pgp-1 glycoprotein concomitant with antigenic stimulation. J Immunol. 1987 May 15;138(10):3120–3129. [PubMed] [Google Scholar]
  2. Coulombel L., Vuillet M. H., Leroy C., Tchernia G. Lineage- and stage-specific adhesion of human hematopoietic progenitor cells to extracellular matrices from marrow fibroblasts. Blood. 1988 Feb;71(2):329–334. [PubMed] [Google Scholar]
  3. Dexter T. M., Allen T. D., Lajtha L. G. Conditions controlling the proliferation of haemopoietic stem cells in vitro. J Cell Physiol. 1977 Jun;91(3):335–344. doi: 10.1002/jcp.1040910303. [DOI] [PubMed] [Google Scholar]
  4. Gallatin M., St John T. P., Siegelman M., Reichert R., Butcher E. C., Weissman I. L. Lymphocyte homing receptors. Cell. 1986 Mar 14;44(5):673–680. doi: 10.1016/0092-8674(86)90832-9. [DOI] [PubMed] [Google Scholar]
  5. Gallatin W. M., Wayner E. A., Hoffman P. A., St John T., Butcher E. C., Carter W. G. Structural homology between lymphocyte receptors for high endothelium and class III extracellular matrix receptor. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4654–4658. doi: 10.1073/pnas.86.12.4654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gimble J. M., Pietrangeli C., Henley A., Dorheim M. A., Silver J., Namen A., Takeichi M., Goridis C., Kincade P. W. Characterization of murine bone marrow and spleen-derived stromal cells: analysis of leukocyte marker and growth factor mRNA transcript levels. Blood. 1989 Jul;74(1):303–311. [PubMed] [Google Scholar]
  7. Goldstein L. A., Zhou D. F., Picker L. J., Minty C. N., Bargatze R. F., Ding J. F., Butcher E. C. A human lymphocyte homing receptor, the hermes antigen, is related to cartilage proteoglycan core and link proteins. Cell. 1989 Mar 24;56(6):1063–1072. doi: 10.1016/0092-8674(89)90639-9. [DOI] [PubMed] [Google Scholar]
  8. Hayashi S., Witte P. L., Shultz L. D., Kincade P. W. Lymphohemopoiesis in culture is prevented by interaction with adherent bone marrow cells from mutant viable motheaten mice. J Immunol. 1988 Apr 1;140(7):2139–2147. [PubMed] [Google Scholar]
  9. Hughes E. N., August J. T. Characterization of plasma membrane proteins identified by monoclonal antibodies. J Biol Chem. 1981 Jan 25;256(2):664–671. [PubMed] [Google Scholar]
  10. Hughes E. N., Colombatti A., August J. T. Murine cell surface glycoproteins. Purification of the polymorphic Pgp-1 antigen and analysis of its expression on macrophages and other myeloid cells. J Biol Chem. 1983 Jan 25;258(2):1014–1021. [PubMed] [Google Scholar]
  11. Hughes E. N., Mengod G., August J. T. Murine cell surface glycoproteins. Characterization of a major component of 80,000 daltons as a polymorphic differentiation antigen of mesenchymal cells. J Biol Chem. 1981 Jul 10;256(13):7023–7027. [PubMed] [Google Scholar]
  12. Ingalls H. M., Goodloe-Holland C. M., Luna E. J. Junctional plasma membrane domains isolated from aggregating Dictyostelium discoideum amebae. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4779–4783. doi: 10.1073/pnas.83.13.4779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jacobson K., O'Dell D., August J. T. Lateral diffusion of an 80,000-dalton glycoprotein in the plasma membrane of murine fibroblasts: relationships to cell structure and function. J Cell Biol. 1984 Nov;99(5):1624–1633. doi: 10.1083/jcb.99.5.1624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jalkanen S., Jalkanen M., Bargatze R., Tammi M., Butcher E. C. Biochemical properties of glycoproteins involved in lymphocyte recognition of high endothelial venules in man. J Immunol. 1988 Sep 1;141(5):1615–1623. [PubMed] [Google Scholar]
  15. Jalkanen S., Wu N., Bargatze R. F., Butcher E. C. Human lymphocyte and lymphoma homing receptors. Annu Rev Med. 1987;38:467–476. doi: 10.1146/annurev.me.38.020187.002343. [DOI] [PubMed] [Google Scholar]
  16. Kincade P. W., Lee G., Pietrangeli C. E., Hayashi S., Gimble J. M. Cells and molecules that regulate B lymphopoiesis in bone marrow. Annu Rev Immunol. 1989;7:111–143. doi: 10.1146/annurev.iy.07.040189.000551. [DOI] [PubMed] [Google Scholar]
  17. Kincade P. W., Lee G., Watanabe T., Sun L., Scheid M. P. Antigens displayed on murine B lymphocyte precursors. J Immunol. 1981 Dec;127(6):2262–2268. [PubMed] [Google Scholar]
  18. Lasky L. A., Singer M. S., Yednock T. A., Dowbenko D., Fennie C., Rodriguez H., Nguyen T., Stachel S., Rosen S. D. Cloning of a lymphocyte homing receptor reveals a lectin domain. Cell. 1989 Mar 24;56(6):1045–1055. doi: 10.1016/0092-8674(89)90637-5. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Letarte M. Human p85 glycoprotein bears three distinct epitopes defined by several monoclonal antibodies. Mol Immunol. 1986 Jun;23(6):639–644. doi: 10.1016/0161-5890(86)90101-x. [DOI] [PubMed] [Google Scholar]
  21. Lewinsohn D. M., Bargatze R. F., Butcher E. C. Leukocyte-endothelial cell recognition: evidence of a common molecular mechanism shared by neutrophils, lymphocytes, and other leukocytes. J Immunol. 1987 Jun 15;138(12):4313–4321. [PubMed] [Google Scholar]
  22. Lynch F., Ceredig R. Ly-24 (Pgp-1) expression by thymocytes and peripheral T cells. Immunol Today. 1988 Jan;9(1):7–10. doi: 10.1016/0167-5699(88)91347-3. [DOI] [PubMed] [Google Scholar]
  23. Miller B. A., Antognetti G., Springer T. A. Identification of cell surface antigens present on murine hematopoietic stem cells. J Immunol. 1985 May;134(5):3286–3290. [PubMed] [Google Scholar]
  24. Murphy T. L., Decker G., August J. T. Glycoproteins of coated pits, cell junctions, and the entire cell surface revealed by monoclonal antibodies and immunoelectron microscopy. J Cell Biol. 1983 Aug;97(2):533–541. doi: 10.1083/jcb.97.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Omary M. B., Trowbridge I. S., Letarte M., Kagnoff M. F., Isacke C. M. Structural heterogeneity of human Pgp-1 and its relationship with p85. Immunogenetics. 1988;27(6):460–464. doi: 10.1007/BF00364434. [DOI] [PubMed] [Google Scholar]
  26. Picker L. J., De los Toyos J., Telen M. J., Haynes B. F., Butcher E. C. Monoclonal antibodies against the CD44 [In(Lu)-related p80], and Pgp-1 antigens in man recognize the Hermes class of lymphocyte homing receptors. J Immunol. 1989 Mar 15;142(6):2046–2051. [PubMed] [Google Scholar]
  27. Siegelman M. H., van de Rijn M., Weissman I. L. Mouse lymph node homing receptor cDNA clone encodes a glycoprotein revealing tandem interaction domains. Science. 1989 Mar 3;243(4895):1165–1172. doi: 10.1126/science.2646713. [DOI] [PubMed] [Google Scholar]
  28. Stamenkovic I., Amiot M., Pesando J. M., Seed B. A lymphocyte molecule implicated in lymph node homing is a member of the cartilage link protein family. Cell. 1989 Mar 24;56(6):1057–1062. doi: 10.1016/0092-8674(89)90638-7. [DOI] [PubMed] [Google Scholar]
  29. Thomas P. S., Pietrangeli C. E., Hayashi S., Schachner M., Goridis C., Low M., Kincade P. W. Demonstration of neural cell adhesion molecules on stromal cells that support lymphopoiesis. Leukemia. 1988 Mar;2(3):171–175. [PubMed] [Google Scholar]
  30. Trowbridge I. S., Lesley J., Schulte R., Hyman R., Trotter J. Biochemical characterization and cellular distribution of a polymorphic, murine cell-surface glycoprotein expressed on lymphoid tissues. Immunogenetics. 1982 Mar;15(3):299–312. doi: 10.1007/BF00364338. [DOI] [PubMed] [Google Scholar]
  31. Trowbridge I. S., Lesley J., Trotter J., Hyman R. Thymocyte subpopulation enriched for progenitors with an unrearranged T-cell receptor beta-chain gene. Nature. 1985 Jun 20;315(6021):666–669. doi: 10.1038/315666a0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]

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