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. 1990 Oct;87(20):8085–8089. doi: 10.1073/pnas.87.20.8085

Expression of CD4 by human megakaryocytes.

R S Basch 1, Y H Kouri 1, S Karpatkin 1
PMCID: PMC54897  PMID: 2236021

Abstract

The CD4 antigen, which serves as the receptor for human immunodeficiency virus type 1 (HIV-1) on T cells, has been detected on human megakaryocytes. Recent evidence of impaired thrombopoiesis in HIV-1-related thrombocytopenia suggested that these cells could be directly infected by the virus and prompted a search for a receptor on megakaryocytes of normal subjects that could permit entry of HIV-1. Bone marrow specimens from uninfected normal control subjects were centrifuged over Ficoll-Hypaque (1.077 g/ml) and analyzed by three-color analysis with a flow cytometer utilizing monoclonal antibodies against CD4 and a glycoprotein present on the surface of megakaryocytes and platelets (GPIIb/IIIa; CD41), as well as 7-aminoactinomycin D, a stain for DNA. Cells presumed to be megakaryocytes were identified by having a DNA content greater than tetraploid and staining brightly with anti-CD41. Approximately 0.4% of the nucleated cells of the marrow met these criteria. Twenty-five percent of these megakaryocytes stained as brightly as CD4+ T cells. Several clones of antibody recognizing different epitopes of the CD4 molecule gave similar results. Platelets were CD4-. Staining of megakaryocytes with anti-CD4 was confirmed by direct microscopic examination of Percoll-gradient-enriched megakaryocytes employing two-color (CD4-phycoerythrin and CD41-fluorescein) immunofluorescence analysis and phase-contrast microscopy. The proportion of double-labeled cells among 112 phase-contrast-identifiable megakaryocytes from five bone marrow specimens varied between 20% and 26% with a mean and SD of 22% +/- 2.5%. Thus some human megakaryocytes express CD4 on their surface that should be capable of binding the HIV-1 gp120 envelope protein. This could serve as a portal of entry for HIV-1.

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Selected References

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

  1. Abrams D. I., Kiprov D. D., Goedert J. J., Sarngadharan M. G., Gallo R. C., Volberding P. A. Antibodies to human T-lymphotropic virus type III and development of the acquired immunodeficiency syndrome in homosexual men presenting with immune thrombocytopenia. Ann Intern Med. 1986 Jan;104(1):47–50. doi: 10.7326/0003-4819-104-1-47. [DOI] [PubMed] [Google Scholar]
  2. Bai Y., Durbin H., Hogg N. Monoclonal antibodies specific for platelet glycoproteins react with human monocytes. Blood. 1984 Jul;64(1):139–146. [PubMed] [Google Scholar]
  3. Ballem P. J., Belzberg A., Devine D., Doubroff P., Mikulash C., Spurston B., Chambers H., Sartori C., Buskard N. Pathophysiology of thrombocytopenia associated with HIV infection in homosexual men. A preliminary report. Blut. 1989 Jul;59(1):111–114. doi: 10.1007/BF00320260. [DOI] [PubMed] [Google Scholar]
  4. Basch R. S., Berman J. W. Thy-1 determinants are present on many murine hematopoietic cells other than T cells. Eur J Immunol. 1982 May;12(5):359–364. doi: 10.1002/eji.1830120502. [DOI] [PubMed] [Google Scholar]
  5. Blakeslee D. Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF). II. Preparation, purity and stability of the compound. J Immunol Methods. 1977;17(3-4):361–364. doi: 10.1016/0022-1759(77)90118-1. [DOI] [PubMed] [Google Scholar]
  6. Dalgleish A. G., Beverley P. C., Clapham P. R., Crawford D. H., Greaves M. F., Weiss R. A. The CD4 (T4) antigen is an essential component of the receptor for the AIDS retrovirus. Nature. 1984 Dec 20;312(5996):763–767. doi: 10.1038/312763a0. [DOI] [PubMed] [Google Scholar]
  7. Dancis A., Ehmann C., Ferziger R., Grima K., Karpatkin S. GPIIIa-related PLA1 antigens with different molecular weights: studies in platelets, endothelial cells, and megakaryocytes. Blood. 1988 Apr;71(4):1056–1061. [PubMed] [Google Scholar]
  8. Deitch A. D., Law H., deVere White R. A stable propidium iodide staining procedure for flow cytometry. J Histochem Cytochem. 1982 Sep;30(9):967–972. doi: 10.1177/30.9.6182188. [DOI] [PubMed] [Google Scholar]
  9. Dialynas D. P., Wilde D. B., Marrack P., Pierres A., Wall K. A., Havran W., Otten G., Loken M. R., Pierres M., Kappler J. Characterization of the murine antigenic determinant, designated L3T4a, recognized by monoclonal antibody GK1.5: expression of L3T4a by functional T cell clones appears to correlate primarily with class II MHC antigen-reactivity. Immunol Rev. 1983;74:29–56. doi: 10.1111/j.1600-065x.1983.tb01083.x. [DOI] [PubMed] [Google Scholar]
  10. Doyle C., Strominger J. L. Interaction between CD4 and class II MHC molecules mediates cell adhesion. Nature. 1987 Nov 19;330(6145):256–259. doi: 10.1038/330256a0. [DOI] [PubMed] [Google Scholar]
  11. Frederickson G. G., Basch R. S. L3T4 antigen expression by hemopoietic precursor cells. J Exp Med. 1989 Apr 1;169(4):1473–1478. doi: 10.1084/jem.169.4.1473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gartner S., Markovits P., Markovitz D. M., Kaplan M. H., Gallo R. C., Popovic M. The role of mononuclear phagocytes in HTLV-III/LAV infection. Science. 1986 Jul 11;233(4760):215–219. doi: 10.1126/science.3014648. [DOI] [PubMed] [Google Scholar]
  13. Gogstad G., Hetland O., Solum N. O., Prydz H. Monocytes and platelets share the glycoproteins IIb and IIIa that are absent from both cells in Glanzmann's thrombasthenia type I. Biochem J. 1983 Aug 15;214(2):331–337. doi: 10.1042/bj2140331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ho D. D., Rota T. R., Hirsch M. S. Infection of monocyte/macrophages by human T lymphotropic virus type III. J Clin Invest. 1986 May;77(5):1712–1715. doi: 10.1172/JCI112491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hymes K. B., Greene J. B., Karpatkin S. The effect of azidothymidine on HIV-related thrombocytopenia. N Engl J Med. 1988 Feb 25;318(8):516–517. doi: 10.1056/NEJM198802253180812. [DOI] [PubMed] [Google Scholar]
  16. Jameson B. A., Rao P. E., Kong L. I., Hahn B. H., Shaw G. M., Hood L. E., Kent S. B. Location and chemical synthesis of a binding site for HIV-1 on the CD4 protein. Science. 1988 Jun 3;240(4857):1335–1339. doi: 10.1126/science.2453925. [DOI] [PubMed] [Google Scholar]
  17. Karpatkin S. Immunologic thrombocytopenic purpura in HIV-seropositive homosexuals, narcotic addicts and hemophiliacs. Semin Hematol. 1988 Jul;25(3):219–229. [PubMed] [Google Scholar]
  18. Klatzmann D., Barré-Sinoussi F., Nugeyre M. T., Danquet C., Vilmer E., Griscelli C., Brun-Veziret F., Rouzioux C., Gluckman J. C., Chermann J. C. Selective tropism of lymphadenopathy associated virus (LAV) for helper-inducer T lymphocytes. Science. 1984 Jul 6;225(4657):59–63. doi: 10.1126/science.6328660. [DOI] [PubMed] [Google Scholar]
  19. Klatzmann D., Champagne E., Chamaret S., Gruest J., Guetard D., Hercend T., Gluckman J. C., Montagnier L. T-lymphocyte T4 molecule behaves as the receptor for human retrovirus LAV. Nature. 1984 Dec 20;312(5996):767–768. doi: 10.1038/312767a0. [DOI] [PubMed] [Google Scholar]
  20. Kurki P., Ogata K., Tan E. M. Monoclonal antibodies to proliferating cell nuclear antigen (PCNA)/cyclin as probes for proliferating cells by immunofluorescence microscopy and flow cytometry. J Immunol Methods. 1988 Apr 22;109(1):49–59. doi: 10.1016/0022-1759(88)90441-3. [DOI] [PubMed] [Google Scholar]
  21. Levine R. F. Isolation and characterization of normal human megakaryocytes. Br J Haematol. 1980 Jul;45(3):487–497. doi: 10.1111/j.1365-2141.1980.tb07168.x. [DOI] [PubMed] [Google Scholar]
  22. Lifson J. D., Feinberg M. B., Reyes G. R., Rabin L., Banapour B., Chakrabarti S., Moss B., Wong-Staal F., Steimer K. S., Engleman E. G. Induction of CD4-dependent cell fusion by the HTLV-III/LAV envelope glycoprotein. Nature. 1986 Oct 23;323(6090):725–728. doi: 10.1038/323725a0. [DOI] [PubMed] [Google Scholar]
  23. Loken M. R., Shah V. O., Dattilio K. L., Civin C. I. Flow cytometric analysis of human bone marrow: I. Normal erythroid development. Blood. 1987 Jan;69(1):255–263. [PubMed] [Google Scholar]
  24. Lucey D. R., Dorsky D. I., Nicholson-Weller A., Weller P. F. Human eosinophils express CD4 protein and bind human immunodeficiency virus 1 gp120. J Exp Med. 1989 Jan 1;169(1):327–332. doi: 10.1084/jem.169.1.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell. 1986 Nov 7;47(3):333–348. doi: 10.1016/0092-8674(86)90590-8. [DOI] [PubMed] [Google Scholar]
  26. Maddon P. J., Littman D. R., Godfrey M., Maddon D. E., Chess L., Axel R. The isolation and nucleotide sequence of a cDNA encoding the T cell surface protein T4: a new member of the immunoglobulin gene family. Cell. 1985 Aug;42(1):93–104. doi: 10.1016/s0092-8674(85)80105-7. [DOI] [PubMed] [Google Scholar]
  27. Marsh M. The entry of enveloped viruses into cells by endocytosis. Biochem J. 1984 Feb 15;218(1):1–10. doi: 10.1042/bj2180001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McDougal J. S., Kennedy M. S., Sligh J. M., Cort S. P., Mawle A., Nicholson J. K. Binding of HTLV-III/LAV to T4+ T cells by a complex of the 110K viral protein and the T4 molecule. Science. 1986 Jan 24;231(4736):382–385. doi: 10.1126/science.3001934. [DOI] [PubMed] [Google Scholar]
  29. Morris L., Distenfeld A., Amorosi E., Karpatkin S. Autoimmune thrombocytopenic purpura in homosexual men. Ann Intern Med. 1982 Jun;96(6 Pt 1):714–717. doi: 10.7326/0003-4819-96-6-714. [DOI] [PubMed] [Google Scholar]
  30. Nicholson J. K., Cross G. D., Callaway C. S., McDougal J. S. In vitro infection of human monocytes with human T lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV). J Immunol. 1986 Jul 1;137(1):323–329. [PubMed] [Google Scholar]
  31. O'Riordan M. L., Berry E. W., Tough I. M. Chromosome studies on bone marrow from a male control population. Br J Haematol. 1970 Jul;19(1):83–90. doi: 10.1111/j.1365-2141.1970.tb01603.x. [DOI] [PubMed] [Google Scholar]
  32. Rabellino E. M., Nachman R. L., Williams N., Winchester R. J., Ross G. D. Human megakaryocytes. I. Characterization of the membrane and cytoplasmic components of isolated marrow megakaryocytes. J Exp Med. 1979 Jun 1;149(6):1273–1287. doi: 10.1084/jem.149.6.1273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rabinovitch P. S., Torres R. M., Engel D. Simultaneous cell cycle analysis and two-color surface immunofluorescence using 7-amino-actinomycin D and single laser excitation: applications to study of cell activation and the cell cycle of murine Ly-1 B cells. J Immunol. 1986 Apr 15;136(8):2769–2775. [PubMed] [Google Scholar]
  34. Ratnoff O. D., Menitove J. E., Aster R. H., Lederman M. M. Coincident classic hemophilia and "idiopathic" thrombocytopenic purpura in patients under treatment with concentrates of antihemophilic factor (factor VIII). N Engl J Med. 1983 Feb 24;308(8):439–442. doi: 10.1056/NEJM198302243080808. [DOI] [PubMed] [Google Scholar]
  35. Reinherz E. L., Kung P. C., Goldstein G., Schlossman S. F. Separation of functional subsets of human T cells by a monoclonal antibody. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4061–4065. doi: 10.1073/pnas.76.8.4061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Savona S., Nardi M. A., Lennette E. T., Karpatkin S. Thrombocytopenic purpura in narcotics addicts. Ann Intern Med. 1985 Jun;102(6):737–741. doi: 10.7326/0003-4819-102-6-737. [DOI] [PubMed] [Google Scholar]
  37. Tabilio A., Pelicci P. G., Vinci G., Mannoni P., Civin C. I., Vainchenker W., Testa U., Lipinski M., Rochant H., Breton-Gorius J. Myeloid and megakaryocytic properties of K-562 cell lines. Cancer Res. 1983 Oct;43(10):4569–4574. [PubMed] [Google Scholar]
  38. Takeda A., Tuazon C. U., Ennis F. A. Antibody-enhanced infection by HIV-1 via Fc receptor-mediated entry. Science. 1988 Oct 28;242(4878):580–583. doi: 10.1126/science.2972065. [DOI] [PubMed] [Google Scholar]
  39. Veillette A., Bookman M. A., Horak E. M., Samelson L. E., Bolen J. B. Signal transduction through the CD4 receptor involves the activation of the internal membrane tyrosine-protein kinase p56lck. Nature. 1989 Mar 16;338(6212):257–259. doi: 10.1038/338257a0. [DOI] [PubMed] [Google Scholar]
  40. Walsh C. M., Nardi M. A., Karpatkin S. On the mechanism of thrombocytopenic purpura in sexually active homosexual men. N Engl J Med. 1984 Sep 6;311(10):635–639. doi: 10.1056/NEJM198409063111004. [DOI] [PubMed] [Google Scholar]
  41. Walsh C., Krigel R., Lennette E., Karpatkin S. Thrombocytopenia in homosexual patients. Prognosis, response to therapy, and prevalence of antibody to the retrovirus associated with the acquired immunodeficiency syndrome. Ann Intern Med. 1985 Oct;103(4):542–545. doi: 10.7326/0003-4819-103-4-542. [DOI] [PubMed] [Google Scholar]
  42. Zucker-Franklin D., Cao Y. Z. Megakaryocytes of human immunodeficiency virus-infected individuals express viral RNA. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5595–5599. doi: 10.1073/pnas.86.14.5595. [DOI] [PMC free article] [PubMed] [Google Scholar]

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