Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1993 May 1;177(5):1331–1342. doi: 10.1084/jem.177.5.1331

Expression of Thy-1 on human hematopoietic progenitor cells

PMCID: PMC2191025  PMID: 7683034

Abstract

Expression of Thy-1 on hematopoietic cells from human fetal liver (FL), cord blood (CB), and bone marrow (BM) was studied with a novel anti-Thy- 1 antibody, 5E10. Specificity of 5E10 for human Thy-1 was demonstrated by immunoprecipitation of a 25-35-kD molecule, and the sequence of a cDNA that was cloned by immunoselection of COS cells transfected with a cDNA library derived from a 5E10+ cell line. Two- and three-color immunofluorescence staining experiments revealed that the Thy-1 expression is restricted to, an average, 1-4% of FL, CB, and BM cells, and binding to these cell types is essentially restricted to a very small subset of lymphoid cells and approximately 25% of CD34+ cells. Thy-1+ CD34+ cells were further characterized as CD38lo/CD45RO+/CD45RA- /CD71lo/c-kit(lo) and rhodamine 123dull. When CD34+ cells were sorted on the basis of Thy-1 expression, the majority of clonogenic cells were recovered in the CD34+Thy-1- fraction, whereas the majority of cells capable of producing myeloid colonies after 5-8 wk of long-term culture (long-term culture initiating cells) were recovered in the Thy-1+CD34+ fraction. In addition to CD34+ cells, Thy-1 was found to be expressed on a variable, very small number (< 1%) of CD34- mononuclear cells in BM, CB, and peripheral blood that were further characterized as CD3+ CD4+ lymphocytes. The restricted expression of Thy-1 on primitive hematopoietic cells is in agreement with a previous report (Baum et al., 1992. Proc. Natl. Acad. Sci. USA. 89:2804) in which Thy-1 expression was used to enrich for primitive hematopoietic cells from fetal tissue. Compared with those previous studies, we found Thy-1 expression on a larger proportion of CD34+ cells (25% in our study vs. 5% in Baum et al.) and furthermore performed studies on Thy-1 expression on CD34+ cells from CB, FL, and BM in relation to markers that are known to be differentially expressed on hematopoietic cells. Taken together our results indicate that Thy-1-specific antibody 5E10 is an attractive tool for further studies on the biology and purification of human stem cells.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Andrews R. G., Singer J. W., Bernstein I. D. Precursors of colony-forming cells in humans can be distinguished from colony-forming cells by expression of the CD33 and CD34 antigens and light scatter properties. J Exp Med. 1989 May 1;169(5):1721–1731. doi: 10.1084/jem.169.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baum C. M., Weissman I. L., Tsukamoto A. S., Buckle A. M., Peault B. Isolation of a candidate human hematopoietic stem-cell population. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2804–2808. doi: 10.1073/pnas.89.7.2804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berman J. W., Basch R. S. Thy-1 antigen expression by murine hematopoietic precursor cells. Exp Hematol. 1985 Dec;13(11):1152–1156. [PubMed] [Google Scholar]
  4. Bourguignon L. Y., Hyman R., Trowbridge I., Singer S. J. Participation of histocompatibility antigens in capping of molecularly independent cell surface components by their specific antibodies. Proc Natl Acad Sci U S A. 1978 May;75(5):2406–2410. doi: 10.1073/pnas.75.5.2406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brandt J., Briddell R. A., Srour E. F., Leemhuis T. B., Hoffman R. Role of c-kit ligand in the expansion of human hematopoietic progenitor cells. Blood. 1992 Feb 1;79(3):634–641. [PubMed] [Google Scholar]
  6. Brandt J., Srour E. F., van Besien K., Briddell R. A., Hoffman R. Cytokine-dependent long-term culture of highly enriched precursors of hematopoietic progenitor cells from human bone marrow. J Clin Invest. 1990 Sep;86(3):932–941. doi: 10.1172/JCI114795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bühring H. J., Asenbauer B., Katrilaka K., Hummel G., Busch F. W. Sequential expression of CD34 and CD33 antigens on myeloid colony-forming cells. Eur J Haematol. 1989 Feb;42(2):143–149. doi: 10.1111/j.1600-0609.1989.tb01203.x. [DOI] [PubMed] [Google Scholar]
  8. Curtis B. M., Williams D. E., Broxmeyer H. E., Dunn J., Farrah T., Jeffery E., Clevenger W., deRoos P., Martin U., Friend D. Enhanced hematopoietic activity of a human granulocyte/macrophage colony-stimulating factor-interleukin 3 fusion protein. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5809–5813. doi: 10.1073/pnas.88.13.5809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  11. Goldschneider I., Metcalf D., Battye F., Mandel T. Analysis of rat hemopoietic cells on the fluorescence-activated cell sorter. I. Isolation of pluripotent hemopoietic stem cells and granulocyte-macrophage progenitor cells. J Exp Med. 1980 Aug 1;152(2):419–437. doi: 10.1084/jem.152.2.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gunter K. C., Malek T. R., Shevach E. M. T cell-activating properties of an anti-Thy-1 monoclonal antibody. Possible analogy to OKT3/Leu-4. J Exp Med. 1984 Mar 1;159(3):716–730. doi: 10.1084/jem.159.3.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. He H. T., Naquet P., Caillol D., Pierres M. Thy-1 supports adhesion of mouse thymocytes to thymic epithelial cells through a Ca2(+)-independent mechanism. J Exp Med. 1991 Feb 1;173(2):515–518. doi: 10.1084/jem.173.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hueber A. O., Pierres M., He H. T. Sulfated glycans directly interact with mouse Thy-1 and negatively regulate Thy-1-mediated adhesion of thymocytes to thymic epithelial cells. J Immunol. 1992 Jun 15;148(12):3692–3699. [PubMed] [Google Scholar]
  15. Ikuta K., Weissman I. L. Evidence that hematopoietic stem cells express mouse c-kit but do not depend on steel factor for their generation. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1502–1506. doi: 10.1073/pnas.89.4.1502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones H. M., Edelman P., Pilkington G. R., Watts M., Linch D. C. Fc gamma RII, but not erythropoietin or GM-CSF, mediates calcium mobilization in fetal hemopoietic blast cells. Exp Hematol. 1992 Mar;20(3):315–319. [PubMed] [Google Scholar]
  17. Kemshead J. T., Heath L., Gibson F. M., Katz F., Richmond F., Treleaven J., Ugelstad J. Magnetic microspheres and monoclonal antibodies for the depletion of neuroblastoma cells from bone marrow: experiences, improvements and observations. Br J Cancer. 1986 Nov;54(5):771–778. doi: 10.1038/bjc.1986.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kroczek R. A., Gunter K. C., Germain R. N., Shevach E. M. Thy-1 functions as a signal transduction molecule in T lymphocytes and transfected B lymphocytes. Nature. 1986 Jul 10;322(6075):181–184. doi: 10.1038/322181a0. [DOI] [PubMed] [Google Scholar]
  19. Kroczek R. A., Gunter K. C., Seligmann B., Shevach E. M. Induction of T cell activation by monoclonal anti-Thy-1 antibodies. J Immunol. 1986 Jun 15;136(12):4379–4384. [PubMed] [Google Scholar]
  20. Lansdorp P. M., Astaldi G. C., Oosterhof F., Janssen M. C., Zeijlemaker W. P. Immunoperoxidase procedures to detect monoclonal antibodies against cell surface antigens. Quantitation of binding and staining of individual cells. J Immunol Methods. 1980;39(4):393–405. doi: 10.1016/0022-1759(80)90240-9. [DOI] [PubMed] [Google Scholar]
  21. Lansdorp P. M., Dragowska W. Long-term erythropoiesis from constant numbers of CD34+ cells in serum-free cultures initiated with highly purified progenitor cells from human bone marrow. J Exp Med. 1992 Jun 1;175(6):1501–1509. doi: 10.1084/jem.175.6.1501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lansdorp P. M., Sutherland H. J., Eaves C. J. Selective expression of CD45 isoforms on functional subpopulations of CD34+ hemopoietic cells from human bone marrow. J Exp Med. 1990 Jul 1;172(1):363–366. doi: 10.1084/jem.172.1.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McKenzie J. L., Fabre J. W. Human thy-1: unusual localization and possible functional significance in lymphoid tissues. J Immunol. 1981 Mar;126(3):843–850. [PubMed] [Google Scholar]
  24. Müller-Sieburg C. E., Tidmarsh G. F., Weissman I. L., Spangrude G. J. Maturation of hematolymphoid cells that express Thy-1. Immunol Ser. 1989;45:289–316. [PubMed] [Google Scholar]
  25. Ostergaard H. L., Shackelford D. A., Hurley T. R., Johnson P., Hyman R., Sefton B. M., Trowbridge I. S. Expression of CD45 alters phosphorylation of the lck-encoded tyrosine protein kinase in murine lymphoma T-cell lines. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8959–8963. doi: 10.1073/pnas.86.22.8959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Papayannopoulou T., Brice M., Yokochi T., Rabinovitch P. S., Lindsley D., Stamatoyannopoulos G. Anti-HEL cell monoclonal antibodies recognize determinants that are also present in hemopoietic progenitors. Blood. 1984 Feb;63(2):326–334. [PubMed] [Google Scholar]
  27. Rutledge R. G., Seligy V. L., Côté M. J., Dimock K., Lewin L. L., Tenniswood M. P. Rapid synthesis and cloning of complementary DNA from any RNA molecule into plasmid and phage M13 vectors. Gene. 1988 Aug 15;68(1):151–158. doi: 10.1016/0378-1119(88)90607-5. [DOI] [PubMed] [Google Scholar]
  28. Seeger R. C., Danon Y. L., Rayner S. A., Hoover F. Definition of a Thy-1 determinant on human neuroblastoma, glioma, sarcoma, and teratoma cells with a monoclonal antibody. J Immunol. 1982 Feb;128(2):983–989. [PubMed] [Google Scholar]
  29. Seki T., Spurr N., Obata F., Goyert S., Goodfellow P., Silver J. The human Thy-1 gene: structure and chromosomal location. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6657–6661. doi: 10.1073/pnas.82.19.6657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Spangrude G. J., Brooks D. M. Phenotypic analysis of mouse hematopoietic stem cells shows a Thy-1-negative subset. Blood. 1992 Oct 15;80(8):1957–1964. [PubMed] [Google Scholar]
  31. Spangrude G. J., Klein J., Heimfeld S., Aihara Y., Weissman I. L. Two monoclonal antibodies identify thymic-repopulating cells in mouse bone marrow. J Immunol. 1989 Jan 15;142(2):425–430. [PubMed] [Google Scholar]
  32. Stefanová I., Horejsí V., Ansotegui I. J., Knapp W., Stockinger H. GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254(5034):1016–1019. doi: 10.1126/science.1719635. [DOI] [PubMed] [Google Scholar]
  33. Sutherland H. J., Eaves C. J., Eaves A. C., Dragowska W., Lansdorp P. M. Characterization and partial purification of human marrow cells capable of initiating long-term hematopoiesis in vitro. Blood. 1989 Oct;74(5):1563–1570. [PubMed] [Google Scholar]
  34. Sutherland H. J., Lansdorp P. M., Henkelman D. H., Eaves A. C., Eaves C. J. Functional characterization of individual human hematopoietic stem cells cultured at limiting dilution on supportive marrow stromal layers. Proc Natl Acad Sci U S A. 1990 May;87(9):3584–3588. doi: 10.1073/pnas.87.9.3584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Terstappen L. W., Huang S., Safford M., Lansdorp P. M., Loken M. R. Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38- progenitor cells. Blood. 1991 Mar 15;77(6):1218–1227. [PubMed] [Google Scholar]
  36. Thomas P. M., Samelson L. E. The glycophosphatidylinositol-anchored Thy-1 molecule interacts with the p60fyn protein tyrosine kinase in T cells. J Biol Chem. 1992 Jun 15;267(17):12317–12322. [PubMed] [Google Scholar]
  37. Till J. E., McCulloch E. A. Hemopoietic stem cell differentiation. Biochim Biophys Acta. 1980 Nov 26;605(4):431–459. doi: 10.1016/0304-419x(80)90009-8. [DOI] [PubMed] [Google Scholar]
  38. Tiveron M. C., Barboni E., Pliego Rivero F. B., Gormley A. M., Seeley P. J., Grosveld F., Morris R. Selective inhibition of neurite outgrowth on mature astrocytes by Thy-1 glycoprotein. Nature. 1992 Feb 20;355(6362):745–748. doi: 10.1038/355745a0. [DOI] [PubMed] [Google Scholar]
  39. Volarević S., Burns C. M., Sussman J. J., Ashwell J. D. Intimate association of Thy-1 and the T-cell antigen receptor with the CD45 tyrosine phosphatase. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7085–7089. doi: 10.1073/pnas.87.18.7085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Williams A. F. Immunoglobulin-related domains for cell surface recognition. Nature. 1985 Apr 18;314(6012):579–580. doi: 10.1038/314579a0. [DOI] [PubMed] [Google Scholar]
  41. Williams D. E., Boswell H. S., Floyd A. D., Broxmeyer H. E. Pluripotential hematopoietic stem cells in post-5-fluorouracil murine bone marrow express the Thy-1 antigen. J Immunol. 1985 Aug;135(2):1004–1011. [PubMed] [Google Scholar]
  42. Williams D. E., Eisenman J., Baird A., Rauch C., Van Ness K., March C. J., Park L. S., Martin U., Mochizuki D. Y., Boswell H. S. Identification of a ligand for the c-kit proto-oncogene. Cell. 1990 Oct 5;63(1):167–174. doi: 10.1016/0092-8674(90)90297-r. [DOI] [PubMed] [Google Scholar]
  43. Wognum A. W., Krystal G., Eaves C. J., Eaves A. C., Lansdorp P. M. Increased erythropoietin-receptor expression on CD34-positive bone marrow cells from patients with chronic myeloid leukemia. Blood. 1992 Feb 1;79(3):642–649. [PubMed] [Google Scholar]

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

RESOURCES