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. 1995 Nov;69(11):6898–6903. doi: 10.1128/jvi.69.11.6898-6903.1995

Functional epitope analysis of the human CD4 molecule: antibodies that inhibit human immunodeficiency virus type 1 gene expression bind to the immunoglobulin CDR3-like region of CD4.

M Benkirane 1, M Hirn 1, D Carrière 1, C Devaux 1
PMCID: PMC189606  PMID: 7474106

Abstract

We recently demonstrated that monoclonal antibody (MAb) 13B8-2, specific for the immunoglobulin (Ig) complementary determining region 3 (CDR3)-like region of the CD4 molecule, inhibits viral transcription in human immunodeficiency virus (HIV)-infected CEM cells and HIV type 1 (HIV-1) promoter activity. Here, we have studied the capacity of several MAb specific for the D1 domain of CD4, including anti-CDR2-like (Leu-3a and ST4) and anti-CDR3-like (13B8-2 and ST40) MAb, and for the D2 domain of CD4 (BL4) to inhibit both provirus transcription in HIV-1LAI-infected CEM cells and transcription of the chloramphenicol acetyltransferase (CAT) gene under control of the HIV-1 long terminal repeat in transiently transfected CEM cells. We found that HIV-1 promoter activity and provirus transcription are inhibited only by MAb that bind to the CDR3-like region in domain 1 of CD4. Moreover, we demonstrated that the Fab fragment of an anti-CDR3-like region-specific anti-CD4 MAb is a powerful inhibitor of HIV-1 promoter activity. These results have implications for understanding the role of the CDR3-like region in CD4 T-cell signaling, which controls provirus transcription.

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Selected References

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  1. Adam D., Klages S., Bishop P., Mahajan S., Escobedo J. A., Bolen J. B. Signal transduction through a biomolecular receptor tyrosine protein kinase composed of a platelet-derived growth factor receptor-CD4 chimera and the nonreceptor tyrosine protein kinase Lck. J Biol Chem. 1993 Sep 15;268(26):19882–19888. [PubMed] [Google Scholar]
  2. Arthos J., Deen K. C., Chaikin M. A., Fornwald J. A., Sathe G., Sattentau Q. J., Clapham P. R., Weiss R. A., McDougal J. S., Pietropaolo C. Identification of the residues in human CD4 critical for the binding of HIV. Cell. 1989 May 5;57(3):469–481. doi: 10.1016/0092-8674(89)90922-7. [DOI] [PubMed] [Google Scholar]
  3. Bank I., Chess L. Perturbation of the T4 molecule transmits a negative signal to T cells. J Exp Med. 1985 Oct 1;162(4):1294–1303. doi: 10.1084/jem.162.4.1294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Batinić D., Robey F. A. The V3 region of the envelope glycoprotein of human immunodeficiency virus type 1 binds sulfated polysaccharides and CD4-derived synthetic peptides. J Biol Chem. 1992 Apr 5;267(10):6664–6671. [PubMed] [Google Scholar]
  5. Benkirane M., Corbeau P., Housset V., Devaux C. An antibody that binds the immunoglobulin CDR3-like region of the CD4 molecule inhibits provirus transcription in HIV-infected T cells. EMBO J. 1993 Dec 15;12(13):4909–4921. doi: 10.1002/j.1460-2075.1993.tb06185.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Benkirane M., Jeang K. T., Devaux C. The cytoplasmic domain of CD4 plays a critical role during the early stages of HIV infection in T-cells. EMBO J. 1994 Dec 1;13(23):5559–5569. doi: 10.1002/j.1460-2075.1994.tb06893.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Benkirane M., Schmid-Antomarchi H., Littman D. R., Hirn M., Rossi B., Devaux C. The cytoplasmic tail of CD4 is required for inhibition of human immunodeficiency virus type 1 replication by antibodies that bind to the immunoglobulin CDR3-like region in domain 1 of CD4. J Virol. 1995 Nov;69(11):6904–6910. doi: 10.1128/jvi.69.11.6904-6910.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Broder C. C., Berger E. A. CD4 molecules with a diversity of mutations encompassing the CDR3 region efficiently support human immunodeficiency virus type 1 envelope glycoprotein-mediated cell fusion. J Virol. 1993 Feb;67(2):913–926. doi: 10.1128/jvi.67.2.913-926.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Camerini D., Seed B. A CD4 domain important for HIV-mediated syncytium formation lies outside the virus binding site. Cell. 1990 Mar 9;60(5):747–754. doi: 10.1016/0092-8674(90)90089-w. [DOI] [PubMed] [Google Scholar]
  10. Chirmule N., Kalyanaraman V. S., Pahwa S. Signals transduced through the CD4 molecule on T lymphocytes activate NF-kappa B. Biochem Biophys Res Commun. 1994 Aug 30;203(1):498–505. doi: 10.1006/bbrc.1994.2210. [DOI] [PubMed] [Google Scholar]
  11. Clayton L. K., Sieh M., Pious D. A., Reinherz E. L. Identification of human CD4 residues affecting class II MHC versus HIV-1 gp120 binding. Nature. 1989 Jun 15;339(6225):548–551. doi: 10.1038/339548a0. [DOI] [PubMed] [Google Scholar]
  12. Corbeau P., Benkirane M., Weil R., David C., Emiliani S., Olive D., Mawas C., Serre A., Devaux C. Ig CDR3-like region of the CD4 molecule is involved in HIV-induced syncytia formation but not in viral entry. J Immunol. 1993 Jan 1;150(1):290–301. [PubMed] [Google Scholar]
  13. Corbeau P., Devaux C., Kourilsky F., Chermann J. C. An early postinfection signal mediated by monoclonal anti-beta 2 microglobulin antibody is responsible for delayed production of human immunodeficiency virus type 1 in peripheral blood mononuclear cells. J Virol. 1990 Apr;64(4):1459–1464. doi: 10.1128/jvi.64.4.1459-1464.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Emmrich F., Kanz L., Eichmann K. Cross-linking of the T cell receptor complex with the subset-specific differentiation antigen stimulates interleukin 2 receptor expression in human CD4 and CD8 T cells. Eur J Immunol. 1987 Apr;17(4):529–534. doi: 10.1002/eji.1830170415. [DOI] [PubMed] [Google Scholar]
  17. Heldin C. H. Dimerization of cell surface receptors in signal transduction. Cell. 1995 Jan 27;80(2):213–223. doi: 10.1016/0092-8674(95)90404-2. [DOI] [PubMed] [Google Scholar]
  18. Hivroz C., Mazerolles F., Soula M., Fagard R., Graton S., Meloche S., Sekaly R. P., Fischer A. Human immunodeficiency virus gp120 and derived peptides activate protein tyrosine kinase p56lck in human CD4 T lymphocytes. Eur J Immunol. 1993 Mar;23(3):600–607. doi: 10.1002/eji.1830230303. [DOI] [PubMed] [Google Scholar]
  19. Jabado N., Le Deist F., Fisher A., Hivroz C. Interaction of HIV gp120 and anti-CD4 antibodies with the CD4 molecule on human CD4+ T cells inhibits the binding activity of NF-AT, NF-kappa B and AP-1, three nuclear factors regulating interleukin-2 gene enhancer activity. Eur J Immunol. 1994 Nov;24(11):2646–2652. doi: 10.1002/eji.1830241112. [DOI] [PubMed] [Google Scholar]
  20. Jameson B. A., McDonnell J. M., Marini J. C., Korngold R. A rationally designed CD4 analogue inhibits experimental allergic encephalomyelitis. Nature. 1994 Apr 21;368(6473):744–746. doi: 10.1038/368744a0. [DOI] [PubMed] [Google Scholar]
  21. Kalyanaraman V. S., Rausch D. M., Osborne J., Padgett M., Hwang K. M., Lifson J. D., Eiden L. E. Evidence by peptide mapping that the region CD4(81-92) is involved in gp120/CD4 interaction leading to HIV infection and HIV-induced syncytium formation. J Immunol. 1990 Dec 15;145(12):4072–4078. [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Kruisbeek A. M., Mond J. J., Fowlkes B. J., Carmen J. A., Bridges S., Longo D. L. Absence of the Lyt-2-,L3T4+ lineage of T cells in mice treated neonatally with anti-I-A correlates with absence of intrathymic I-A-bearing antigen-presenting cell function. J Exp Med. 1985 May 1;161(5):1029–1047. doi: 10.1084/jem.161.5.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lamarre D., Capon D. J., Karp D. R., Gregory T., Long E. O., Sékaly R. P. Class II MHC molecules and the HIV gp 120 envelope protein interact with functionally distinct regions of the CD4 molecule. EMBO J. 1989 Nov;8(11):3271–3277. doi: 10.1002/j.1460-2075.1989.tb08487.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Langedijk J. P., Puijk W. C., van Hoorn W. P., Meloen R. H. Location of CD4 dimerization site explains critical role of CDR3-like region in HIV-1 infection and T-cell activation and implies a model for complex of coreceptor-MHC. J Biol Chem. 1993 Aug 15;268(23):16875–16878. [PubMed] [Google Scholar]
  27. Ledbetter J. A., June C. H., Rabinovitch P. S., Grossmann A., Tsu T. T., Imboden J. B. Signal transduction through CD4 receptors: stimulatory vs. inhibitory activity is regulated by CD4 proximity to the CD3/T cell receptor. Eur J Immunol. 1988 Apr;18(4):525–532. doi: 10.1002/eji.1830180406. [DOI] [PubMed] [Google Scholar]
  28. Lifson J. D., Hwang K. M., Nara P. L., Fraser B., Padgett M., Dunlop N. M., Eiden L. E. Synthetic CD4 peptide derivatives that inhibit HIV infection and cytopathicity. Science. 1988 Aug 5;241(4866):712–716. doi: 10.1126/science.2969619. [DOI] [PubMed] [Google Scholar]
  29. Littman D. R. The structure of the CD4 and CD8 genes. Annu Rev Immunol. 1987;5:561–584. doi: 10.1146/annurev.iy.05.040187.003021. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. McDonnell J. M., Blank K. J., Rao P. E., Jameson B. A. Direct involvement of the CDR3-like domain of CD4 in T helper cell activation. J Immunol. 1992 Sep 1;149(5):1626–1630. [PubMed] [Google Scholar]
  32. Mizukami T., Fuerst T. R., Berger E. A., Moss B. Binding region for human immunodeficiency virus (HIV) and epitopes for HIV-blocking monoclonal antibodies of the CD4 molecule defined by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9273–9277. doi: 10.1073/pnas.85.23.9273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nara P. L., Hwang K. M., Rausch D. M., Lifson J. D., Eiden L. E. CD4 antigen-based antireceptor peptides inhibit infectivity of human immunodeficiency virus in vitro at multiple stages of the viral life cycle. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7139–7143. doi: 10.1073/pnas.86.18.7139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ohki K., Kimura T., Ohmura K., Kato S., Ikuta K. Blocking of HIV-1 infection, but not HIV-1-induced syncytium formation, by a CD4 peptide derivative partly corresponding to an immunoglobulin CDR3. AIDS. 1990 Nov;4(11):1160–1161. doi: 10.1097/00002030-199011000-00020. [DOI] [PubMed] [Google Scholar]
  35. Peterson A., Seed B. Genetic analysis of monoclonal antibody and HIV binding sites on the human lymphocyte antigen CD4. Cell. 1988 Jul 1;54(1):65–72. doi: 10.1016/0092-8674(88)90180-8. [DOI] [PubMed] [Google Scholar]
  36. Rausch D. M., Hwang K. M., Padgett M., Voltz A. H., Rivas A., Engleman E., Gaston I., McGrath M., Fraser B., Kalyanaraman V. S. Peptides derived from the CDR3-homologous domain of the CD4 molecule are specific inhibitors of HIV-1 and SIV infection, virus-induced cell fusion, and postinfection viral transmission in vitro. Implications for the design of small peptide anti-HIV therapeutic agents. Ann N Y Acad Sci. 1990;616:125–148. doi: 10.1111/j.1749-6632.1990.tb17834.x. [DOI] [PubMed] [Google Scholar]
  37. Repke H., Gabuzda D., Palù G., Emmrich F., Sodroski J. Effects of CD4 synthetic peptides on HIV type I envelope glycoprotein function. J Immunol. 1992 Sep 1;149(5):1809–1816. [PubMed] [Google Scholar]
  38. Robert V., Resnicoff M., Chermann J. C., Devaux C. Characterization of monoclonal antibodies identifying type and strain-specific epitopes of human immunodeficiency virus type 1. Mol Cell Biochem. 1991 Apr 10;102(2):115–123. doi: 10.1007/BF00234569. [DOI] [PubMed] [Google Scholar]
  39. Rosoff P. M., Burakoff S. J., Greenstein J. L. The role of the L3T4 molecule in mitogen and antigen-activated signal transduction. Cell. 1987 Jun 19;49(6):845–853. doi: 10.1016/0092-8674(87)90622-2. [DOI] [PubMed] [Google Scholar]
  40. Rudd C. E., Trevillyan J. M., Dasgupta J. D., Wong L. L., Schlossman S. F. The CD4 receptor is complexed in detergent lysates to a protein-tyrosine kinase (pp58) from human T lymphocytes. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5190–5194. doi: 10.1073/pnas.85.14.5190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ryu S. E., Kwong P. D., Truneh A., Porter T. G., Arthos J., Rosenberg M., Dai X. P., Xuong N. H., Axel R., Sweet R. W. Crystal structure of an HIV-binding recombinant fragment of human CD4. Nature. 1990 Nov 29;348(6300):419–426. doi: 10.1038/348419a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Saizawa K., Rojo J., Janeway C. A., Jr Evidence for a physical association of CD4 and the CD3:alpha:beta T-cell receptor. Nature. 1987 Jul 16;328(6127):260–263. doi: 10.1038/328260a0. [DOI] [PubMed] [Google Scholar]
  43. Sattentau Q. J., Arthos J., Deen K., Hanna N., Healey D., Beverley P. C., Sweet R., Truneh A. Structural analysis of the human immunodeficiency virus-binding domain of CD4. Epitope mapping with site-directed mutants and anti-idiotypes. J Exp Med. 1989 Oct 1;170(4):1319–1334. doi: 10.1084/jem.170.4.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sattentau Q. J., Dalgleish A. G., Weiss R. A., Beverley P. C. Epitopes of the CD4 antigen and HIV infection. Science. 1986 Nov 28;234(4780):1120–1123. doi: 10.1126/science.2430333. [DOI] [PubMed] [Google Scholar]
  45. Simon J. H., Somoza C., Schockmel G. A., Collin M., Davis S. J., Williams A. F., James W. A rat CD4 mutant containing the gp120-binding site mediates human immunodeficiency virus type 1 infection. J Exp Med. 1993 Apr 1;177(4):949–954. doi: 10.1084/jem.177.4.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Truneh A., Buck D., Cassatt D. R., Juszczak R., Kassis S., Ryu S. E., Healey D., Sweet R., Sattentau Q. A region in domain 1 of CD4 distinct from the primary gp120 binding site is involved in HIV infection and virus-mediated fusion. J Biol Chem. 1991 Mar 25;266(9):5942–5948. [PubMed] [Google Scholar]
  47. 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]
  48. Wang J. H., Yan Y. W., Garrett T. P., Liu J. H., Rodgers D. W., Garlick R. L., Tarr G. E., Husain Y., Reinherz E. L., Harrison S. C. Atomic structure of a fragment of human CD4 containing two immunoglobulin-like domains. Nature. 1990 Nov 29;348(6300):411–418. doi: 10.1038/348411a0. [DOI] [PubMed] [Google Scholar]
  49. Wassmer P., Chan C., Lögdberg L., Shevach E. M. Role of the L3T4-antigen in T cell activation. II. Inhibition of T cell activation by monoclonal anti-L3T4 antibodies in the absence of accessory cells. J Immunol. 1985 Oct;135(4):2237–2242. [PubMed] [Google Scholar]

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