Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1995 Jul;69(7):4045–4052. doi: 10.1128/jvi.69.7.4045-4052.1995

Inhibition of clinical human immunodeficiency virus (HIV) type 1 isolates in primary CD4+ T lymphocytes by retroviral vectors expressing anti-HIV genes.

T Vandendriessche 1, M K Chuah 1, L Chiang 1, H K Chang 1, B Ensoli 1, R A Morgan 1
PMCID: PMC189138  PMID: 7769662

Abstract

Gene therapy may be of benefit in human immunodeficiency virus type 1 (HIV-1)-infected individuals by virtue of its ability to inhibit virus replication and prevent viral gene expression. It is not known whether anti-HIV-1 gene therapy strategies based on antisense or transdominant HIV-1 mutant proteins can inhibit the replication and expression of clinical HIV-1 isolates in primary CD4+ T lymphocytes. We therefore transduced CD4+ T lymphocytes from uninfected individuals with retroviral vectors expressing either HIV-1-specific antisense-TAR or antisense-Tat/Rev RNA, transdominant HIV-1 Rev protein, and a combination of antisense-TAR and transdominant Rev. The engineered CD4+ T lymphocytes were then infected with four different clinical HIV-1 isolates. We found that replication of all HIV-1 isolates was inhibited by all the anti-HIV vectors tested. Greater inhibition of HIV-1 was observed with transdominant Rev than with antisense RNA. We hereby demonstrated effective protection by antisense RNA or transdominant mutant proteins against HIV-1 infection in primary CD4+ T lymphocytes using clinical HIV-1 isolates, and this represents an essential step toward clinical anti-HIV-1 gene therapy.

Full Text

The Full Text of this article is available as a PDF (247.3 KB).

Selected References

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

  1. Adeniyi-Jones S., Romeo P. H., Zasloff M. Generation of long read-through transcripts in vivo and in vitro by deletion of 3' termination and processing sequences in the human tRNAimet gene. Nucleic Acids Res. 1984 Jan 25;12(2):1101–1115. doi: 10.1093/nar/12.2.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson W. French, Blaese R. Michael, Culver Kenneth. The ADA human gene therapy clinical protocol: Points to Consider response with clinical protocol, July 6, 1990. Hum Gene Ther. 1990 Fall;1(3):331–362. doi: 10.1089/hum.1990.1.3-331. [DOI] [PubMed] [Google Scholar]
  3. Bahner I., Zhou C., Yu X. J., Hao Q. L., Guatelli J. C., Kohn D. B. Comparison of trans-dominant inhibitory mutant human immunodeficiency virus type 1 genes expressed by retroviral vectors in human T lymphocytes. J Virol. 1993 Jun;67(6):3199–3207. doi: 10.1128/jvi.67.6.3199-3207.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Banda N. K., Bernier J., Kurahara D. K., Kurrle R., Haigwood N., Sekaly R. P., Finkel T. H. Crosslinking CD4 by human immunodeficiency virus gp120 primes T cells for activation-induced apoptosis. J Exp Med. 1992 Oct 1;176(4):1099–1106. doi: 10.1084/jem.176.4.1099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berkhout B., Silverman R. H., Jeang K. T. Tat trans-activates the human immunodeficiency virus through a nascent RNA target. Cell. 1989 Oct 20;59(2):273–282. doi: 10.1016/0092-8674(89)90289-4. [DOI] [PubMed] [Google Scholar]
  6. Bodine D. M., McDonagh K. T., Brandt S. J., Ney P. A., Agricola B., Byrne E., Nienhuis A. W. Development of a high-titer retrovirus producer cell line capable of gene transfer into rhesus monkey hematopoietic stem cells. Proc Natl Acad Sci U S A. 1990 May;87(10):3738–3742. doi: 10.1073/pnas.87.10.3738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chatterjee S., Johnson P. R., Wong K. K., Jr Dual-target inhibition of HIV-1 in vitro by means of an adeno-associated virus antisense vector. Science. 1992 Nov 27;258(5087):1485–1488. doi: 10.1126/science.1359646. [DOI] [PubMed] [Google Scholar]
  8. Chuah M. K., VandenDriessche T., Chang H. K., Ensoli B., Morgan R. A. Inhibition of human immunodeficiency virus type-1 by retroviral vectors expressing antisense-TAR. Hum Gene Ther. 1994 Dec;5(12):1467–1475. doi: 10.1089/hum.1994.5.12-1467. [DOI] [PubMed] [Google Scholar]
  9. Cullen B. R. Does HIV-1 Tat induce a change in viral initiation rights? Cell. 1993 May 7;73(3):417–420. doi: 10.1016/0092-8674(93)90126-b. [DOI] [PubMed] [Google Scholar]
  10. Culver K. W., Morgan R. A., Osborne W. R., Lee R. T., Lenschow D., Able C., Cornetta K., Anderson W. F., Blaese R. M. In vivo expression and survival of gene-modified T lymphocytes in rhesus monkeys. Hum Gene Ther. 1990 Winter;1(4):399–410. doi: 10.1089/hum.1990.1.4-399. [DOI] [PubMed] [Google Scholar]
  11. Culver K., Cornetta K., Morgan R., Morecki S., Aebersold P., Kasid A., Lotze M., Rosenberg S. A., Anderson W. F., Blaese R. M. Lymphocytes as cellular vehicles for gene therapy in mouse and man. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3155–3159. doi: 10.1073/pnas.88.8.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Embretson J., Zupancic M., Ribas J. L., Burke A., Racz P., Tenner-Racz K., Haase A. T. Massive covert infection of helper T lymphocytes and macrophages by HIV during the incubation period of AIDS. Nature. 1993 Mar 25;362(6418):359–362. doi: 10.1038/362359a0. [DOI] [PubMed] [Google Scholar]
  13. Ensoli B., Barillari G., Salahuddin S. Z., Gallo R. C., Wong-Staal F. Tat protein of HIV-1 stimulates growth of cells derived from Kaposi's sarcoma lesions of AIDS patients. Nature. 1990 May 3;345(6270):84–86. doi: 10.1038/345084a0. [DOI] [PubMed] [Google Scholar]
  14. Fischer U., Meyer S., Teufel M., Heckel C., Lührmann R., Rautmann G. Evidence that HIV-1 Rev directly promotes the nuclear export of unspliced RNA. EMBO J. 1994 Sep 1;13(17):4105–4112. doi: 10.1002/j.1460-2075.1994.tb06728.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Frankel A. D. Activation of HIV transcription by Tat. Curr Opin Genet Dev. 1992 Apr;2(2):293–298. doi: 10.1016/s0959-437x(05)80287-4. [DOI] [PubMed] [Google Scholar]
  16. Gilboa E., Smith C. Gene therapy for infectious diseases: the AIDS model. Trends Genet. 1994 Apr;10(4):139–144. doi: 10.1016/0168-9525(94)90216-x. [DOI] [PubMed] [Google Scholar]
  17. Gurgo C., Guo H. G., Franchini G., Aldovini A., Collalti E., Farrell K., Wong-Staal F., Gallo R. C., Reitz M. S., Jr Envelope sequences of two new United States HIV-1 isolates. Virology. 1988 Jun;164(2):531–536. doi: 10.1016/0042-6822(88)90568-5. [DOI] [PubMed] [Google Scholar]
  18. Hill J. M., Mervis R. F., Avidor R., Moody T. W., Brenneman D. E. HIV envelope protein-induced neuronal damage and retardation of behavioral development in rat neonates. Brain Res. 1993 Feb 19;603(2):222–233. doi: 10.1016/0006-8993(93)91241-j. [DOI] [PubMed] [Google Scholar]
  19. Hope T. J., Klein N. P., Elder M. E., Parslow T. G. trans-dominant inhibition of human immunodeficiency virus type 1 Rev occurs through formation of inactive protein complexes. J Virol. 1992 Apr;66(4):1849–1855. doi: 10.1128/jvi.66.4.1849-1855.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kalland K. H., Szilvay A. M., Brokstad K. A., Saetrevik W., Haukenes G. The human immunodeficiency virus type 1 Rev protein shuttles between the cytoplasm and nuclear compartments. Mol Cell Biol. 1994 Nov;14(11):7436–7444. doi: 10.1128/mcb.14.11.7436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laspia M. F., Rice A. P., Mathews M. B. HIV-1 Tat protein increases transcriptional initiation and stabilizes elongation. Cell. 1989 Oct 20;59(2):283–292. doi: 10.1016/0092-8674(89)90290-0. [DOI] [PubMed] [Google Scholar]
  22. Laurence J., Astrin S. M. Human immunodeficiency virus induction of malignant transformation in human B lymphocytes. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7635–7639. doi: 10.1073/pnas.88.17.7635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leavitt M. C., Yu M., Yamada O., Kraus G., Looney D., Poeschla E., Wong-Staal F. Transfer of an anti-HIV-1 ribozyme gene into primary human lymphocytes. Hum Gene Ther. 1994 Sep;5(9):1115–1120. doi: 10.1089/hum.1994.5.9-1115. [DOI] [PubMed] [Google Scholar]
  24. Lee S. W., Gallardo H. F., Gilboa E., Smith C. Inhibition of human immunodeficiency virus type 1 in human T cells by a potent Rev response element decoy consisting of the 13-nucleotide minimal Rev-binding domain. J Virol. 1994 Dec;68(12):8254–8264. doi: 10.1128/jvi.68.12.8254-8264.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lee T. C., Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of RRE-derived sequences inhibits HIV-1 replication in CEM cells. New Biol. 1992 Jan;4(1):66–74. [PubMed] [Google Scholar]
  26. Li Y., Kappes J. C., Conway J. A., Price R. W., Shaw G. M., Hahn B. H. Molecular characterization of human immunodeficiency virus type 1 cloned directly from uncultured human brain tissue: identification of replication-competent and -defective viral genomes. J Virol. 1991 Aug;65(8):3973–3985. doi: 10.1128/jvi.65.8.3973-3985.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Liu J., Woffendin C., Yang Z. Y., Nabel G. J. Regulated expression of a dominant negative form of Rev improves resistance to HIV replication in T cells. Gene Ther. 1994 Jan;1(1):32–37. [PubMed] [Google Scholar]
  28. Lu Y. Y., Koga Y., Tanaka K., Sasaki M., Kimura G., Nomoto K. Apoptosis induced in CD4+ cells expressing gp160 of human immunodeficiency virus type 1. J Virol. 1994 Jan;68(1):390–399. doi: 10.1128/jvi.68.1.390-399.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Malim M. H., Böhnlein S., Hauber J., Cullen B. R. Functional dissection of the HIV-1 Rev trans-activator--derivation of a trans-dominant repressor of Rev function. Cell. 1989 Jul 14;58(1):205–214. doi: 10.1016/0092-8674(89)90416-9. [DOI] [PubMed] [Google Scholar]
  30. Malim M. H., Cullen B. R. Rev and the fate of pre-mRNA in the nucleus: implications for the regulation of RNA processing in eukaryotes. Mol Cell Biol. 1993 Oct;13(10):6180–6189. doi: 10.1128/mcb.13.10.6180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Malim M. H., Freimuth W. W., Liu J., Boyle T. J., Lyerly H. K., Cullen B. R., Nabel G. J. Stable expression of transdominant Rev protein in human T cells inhibits human immunodeficiency virus replication. J Exp Med. 1992 Oct 1;176(4):1197–1201. doi: 10.1084/jem.176.4.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Markowitz D., Goff S., Bank A. A safe packaging line for gene transfer: separating viral genes on two different plasmids. J Virol. 1988 Apr;62(4):1120–1124. doi: 10.1128/jvi.62.4.1120-1124.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McLachlin J. R., Mittereder N., Daucher M. B., Kadan M., Eglitis M. A. Factors affecting retroviral vector function and structural integrity. Virology. 1993 Jul;195(1):1–5. doi: 10.1006/viro.1993.1340. [DOI] [PubMed] [Google Scholar]
  34. Miller A. D., Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol. 1986 Aug;6(8):2895–2902. doi: 10.1128/mcb.6.8.2895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miller S. B., Tse H., Rosenspire A. J., King S. R. CD4-independent inhibition of lymphocyte proliferation mediated by HIV-1 envelope glycoproteins. Virology. 1992 Dec;191(2):973–977. doi: 10.1016/0042-6822(92)90275-t. [DOI] [PubMed] [Google Scholar]
  36. Muesing M. A., Smith D. H., Capon D. J. Regulation of mRNA accumulation by a human immunodeficiency virus trans-activator protein. Cell. 1987 Feb 27;48(4):691–701. doi: 10.1016/0092-8674(87)90247-9. [DOI] [PubMed] [Google Scholar]
  37. Ojwang J. O., Hampel A., Looney D. J., Wong-Staal F., Rappaport J. Inhibition of human immunodeficiency virus type 1 expression by a hairpin ribozyme. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10802–10806. doi: 10.1073/pnas.89.22.10802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Orloff S. L., Kennedy M. S., Belperron A. A., Maddon P. J., McDougal J. S. Two mechanisms of soluble CD4 (sCD4)-mediated inhibition of human immunodeficiency virus type 1 (HIV-1) infectivity and their relation to primary HIV-1 isolates with reduced sensitivity to sCD4. J Virol. 1993 Mar;67(3):1461–1471. doi: 10.1128/jvi.67.3.1461-1471.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Pantaleo G., Graziosi C., Demarest J. F., Butini L., Montroni M., Fox C. H., Orenstein J. M., Kotler D. P., Fauci A. S. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease. Nature. 1993 Mar 25;362(6418):355–358. doi: 10.1038/362355a0. [DOI] [PubMed] [Google Scholar]
  40. Richman D. D. HIV drug resistance. AIDS Res Hum Retroviruses. 1992 Jun;8(6):1065–1071. doi: 10.1089/aid.1992.8.1065. [DOI] [PubMed] [Google Scholar]
  41. Sabatier J. M., Vives E., Mabrouk K., Benjouad A., Rochat H., Duval A., Hue B., Bahraoui E. Evidence for neurotoxic activity of tat from human immunodeficiency virus type 1. J Virol. 1991 Feb;65(2):961–967. doi: 10.1128/jvi.65.2.961-967.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sarver N., Cantin E. M., Chang P. S., Zaia J. A., Ladne P. A., Stephens D. A., Rossi J. J. Ribozymes as potential anti-HIV-1 therapeutic agents. Science. 1990 Mar 9;247(4947):1222–1225. doi: 10.1126/science.2107573. [DOI] [PubMed] [Google Scholar]
  43. Sczakiel G., Oppenländer M., Rittner K., Pawlita M. Tat- and Rev-directed antisense RNA expression inhibits and abolishes replication of human immunodeficiency virus type 1: a temporal analysis. J Virol. 1992 Sep;66(9):5576–5581. doi: 10.1128/jvi.66.9.5576-5581.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replication. Cell. 1990 Nov 2;63(3):601–608. doi: 10.1016/0092-8674(90)90455-n. [DOI] [PubMed] [Google Scholar]
  45. Temin H. M., Bolognesi D. P. AIDS. Where has HIV been hiding? Nature. 1993 Mar 25;362(6418):292–293. doi: 10.1038/362292a0. [DOI] [PubMed] [Google Scholar]
  46. Vaishnav Y. N., Wong-Staal F. The biochemistry of AIDS. Annu Rev Biochem. 1991;60:577–630. doi: 10.1146/annurev.bi.60.070191.003045. [DOI] [PubMed] [Google Scholar]
  47. Walker R., Blaese R. M., Carter C. S., Chang L., Klein H., Lane H. C., Leitman S. F., Mullen C. A., Larson M. A study of the safety and survival of the adoptive transfer of genetically marked syngeneic lymphocytes in HIV-infected identical twins. Hum Gene Ther. 1993 Oct;4(5):659–680. doi: 10.1089/hum.1993.4.5-659. [DOI] [PubMed] [Google Scholar]
  48. Yu M., Ojwang J., Yamada O., Hampel A., Rapapport J., Looney D., Wong-Staal F. A hairpin ribozyme inhibits expression of diverse strains of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6340–6344. doi: 10.1073/pnas.90.13.6340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yu M., Poeschla E., Wong-Staal F. Progress towards gene therapy for HIV infection. Gene Ther. 1994 Jan;1(1):13–26. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES