Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Aug 15;100(4):893–903. doi: 10.1172/JCI119605

Dynamics of HIV-1 mRNA expression in patients with long-term nonprogressive HIV-1 infection.

M Comar 1, C Simonelli 1, S Zanussi 1, P Paoli 1, E Vaccher 1, U Tirelli 1, M Giacca 1
PMCID: PMC508262  PMID: 9259589

Abstract

A large number of evidences indicate that progression of HIV disease is driven by an increase in viral burden. It is still unclear, however, to what extent this is contributed by the dysregulation of the molecular mechanisms governing virus gene expression at the transcriptional or posttranscriptional levels. To address this issue, several quantitative virologic parameters (including provirus transcriptional activity and splicing pattern) were analyzed in individuals with nonprogressive HIV infection and compared with those of a matched group of progressor patients. Exact quantification was achieved by a competitive PCR procedure using a multicompetitor template. Nonprogressors were characterized by striking differences in the levels of viremia, provirus copy number, and overall levels of all viral mRNA classes in peripheral blood mononuclear cells. Additionally, the transcriptional activity of the proviral DNA in these patients was mainly engaged in the production of multiprocessed transcripts, with a pattern resembling the early phases of the experimental infection. Taken together, these results show that both viral load and provirus transcription pattern are remarkably different in infected individuals nonprogressing toward overt disease, and further support the notion that disease progression is accompanied by a change in the kinetics of HIV gene expression.

Full Text

The Full Text of this article is available as a PDF (360.6 KB).

Selected References

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

  1. Bagasra O., Hauptman S. P., Lischner H. W., Sachs M., Pomerantz R. J. Detection of human immunodeficiency virus type 1 provirus in mononuclear cells by in situ polymerase chain reaction. N Engl J Med. 1992 May 21;326(21):1385–1391. doi: 10.1056/NEJM199205213262103. [DOI] [PubMed] [Google Scholar]
  2. Bagnarelli P., Menzo S., Valenza A., Manzin A., Giacca M., Ancarani F., Scalise G., Varaldo P. E., Clementi M. Molecular profile of human immunodeficiency virus type 1 infection in symptomless patients and in patients with AIDS. J Virol. 1992 Dec;66(12):7328–7335. doi: 10.1128/jvi.66.12.7328-7335.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bagnarelli P., Valenza A., Menzo S., Manzin A., Scalise G., Varaldo P. E., Clementi M. Dynamics of molecular parameters of human immunodeficiency virus type 1 activity in vivo. J Virol. 1994 Apr;68(4):2495–2502. doi: 10.1128/jvi.68.4.2495-2502.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buchbinder S. P., Katz M. H., Hessol N. A., O'Malley P. M., Holmberg S. D. Long-term HIV-1 infection without immunologic progression. AIDS. 1994 Aug;8(8):1123–1128. doi: 10.1097/00002030-199408000-00014. [DOI] [PubMed] [Google Scholar]
  5. Cao Y., Qin L., Zhang L., Safrit J., Ho D. D. Virologic and immunologic characterization of long-term survivors of human immunodeficiency virus type 1 infection. N Engl J Med. 1995 Jan 26;332(4):201–208. doi: 10.1056/NEJM199501263320401. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Chun T. W., Finzi D., Margolick J., Chadwick K., Schwartz D., Siliciano R. F. In vivo fate of HIV-1-infected T cells: quantitative analysis of the transition to stable latency. Nat Med. 1995 Dec;1(12):1284–1290. doi: 10.1038/nm1295-1284. [DOI] [PubMed] [Google Scholar]
  8. Comar M., Marzio G., D'Agaro P., Giacca M. Quantitative dynamics of HIV type 1 expression. AIDS Res Hum Retroviruses. 1996 Jan 20;12(2):117–126. doi: 10.1089/aid.1996.12.117. [DOI] [PubMed] [Google Scholar]
  9. Connor R. I., Mohri H., Cao Y., Ho D. D. Increased viral burden and cytopathicity correlate temporally with CD4+ T-lymphocyte decline and clinical progression in human immunodeficiency virus type 1-infected individuals. J Virol. 1993 Apr;67(4):1772–1777. doi: 10.1128/jvi.67.4.1772-1777.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cullen B. R., Greene W. C. Regulatory pathways governing HIV-1 replication. Cell. 1989 Aug 11;58(3):423–426. doi: 10.1016/0092-8674(89)90420-0. [DOI] [PubMed] [Google Scholar]
  11. De Rossi A., Masiero S., Giaquinto C., Ruga E., Comar M., Giacca M., Chieco-Bianchi L. Dynamics of viral replication in infants with vertically acquired human immunodeficiency virus type 1 infection. J Clin Invest. 1996 Jan 15;97(2):323–330. doi: 10.1172/JCI118419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Demarchi F., D'Agaro P., Falaschi A., Giacca M. In vivo footprinting analysis of constitutive and inducible protein-DNA interactions at the long terminal repeat of human immunodeficiency virus type 1. J Virol. 1993 Dec;67(12):7450–7460. doi: 10.1128/jvi.67.12.7450-7460.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dianzani F., Antonelli G., Riva E., Uccini S., Visco G. Plasma HIV viremia and viral load in lymph nodes. Nat Med. 1996 Aug;2(8):832–833. doi: 10.1038/nm0896-832. [DOI] [PubMed] [Google Scholar]
  14. Diviacco S., Norio P., Zentilin L., Menzo S., Clementi M., Biamonti G., Riva S., Falaschi A., Giacca M. A novel procedure for quantitative polymerase chain reaction by coamplification of competitive templates. Gene. 1992 Dec 15;122(2):313–320. doi: 10.1016/0378-1119(92)90220-j. [DOI] [PubMed] [Google Scholar]
  15. Easterbrook P. J. Non-progression in HIV infection. AIDS. 1994 Aug;8(8):1179–1182. doi: 10.1097/00002030-199408000-00023. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Furtado M. R., Kingsley L. A., Wolinsky S. M. Changes in the viral mRNA expression pattern correlate with a rapid rate of CD4+ T-cell number decline in human immunodeficiency virus type 1-infected individuals. J Virol. 1995 Apr;69(4):2092–2100. doi: 10.1128/jvi.69.4.2092-2100.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Giacca M., Zanussi S., Comar M., Simonelli C., Vaccher E., de Paoli P., Tirelli U. Treatment of human immunodeficiency virus infection with hydroxyurea: virologic and clinical evaluation. J Infect Dis. 1996 Jul;174(1):204–209. doi: 10.1093/infdis/174.1.204. [DOI] [PubMed] [Google Scholar]
  19. Giacca M., Zentilin L., Norio P., Diviacco S., Dimitrova D., Contreas G., Biamonti G., Perini G., Weighardt F., Riva S. Fine mapping of a replication origin of human DNA. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7119–7123. doi: 10.1073/pnas.91.15.7119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Grassi G., Pozzato G., Moretti M., Giacca M. Quantitative analysis of hepatitis C virus RNA in liver biopsies by competitive reverse transcription and polymerase chain reaction. J Hepatol. 1995 Oct;23(4):403–411. doi: 10.1016/0168-8278(95)80198-7. [DOI] [PubMed] [Google Scholar]
  21. Grassi G., Zentilin L., Tafuro S., Diviacco S., Ventura A., Falaschi A., Giacca M. A rapid procedure for the quantitation of low abundance RNAs by competitive reverse transcription-polymerase chain reaction. Nucleic Acids Res. 1994 Oct 25;22(21):4547–4549. doi: 10.1093/nar/22.21.4547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Greenough T. C., Somasundaran M., Brettler D. B., Hesselton R. M., Alimenti A., Kirchhoff F., Panicali D., Sullivan J. L. Normal immune function and inability to isolate virus in culture in an individual with long-term human immunodeficiency virus type 1 infection. AIDS Res Hum Retroviruses. 1994 Apr;10(4):395–403. doi: 10.1089/aid.1994.10.395. [DOI] [PubMed] [Google Scholar]
  23. Gupta P., Kingsley L., Armstrong J., Ding M., Cottrill M., Rinaldo C. Enhanced expression of human immunodeficiency virus type 1 correlates with development of AIDS. Virology. 1993 Oct;196(2):586–595. doi: 10.1006/viro.1993.1514. [DOI] [PubMed] [Google Scholar]
  24. Ho D. D., Moudgil T., Alam M. Quantitation of human immunodeficiency virus type 1 in the blood of infected persons. N Engl J Med. 1989 Dec 14;321(24):1621–1625. doi: 10.1056/NEJM198912143212401. [DOI] [PubMed] [Google Scholar]
  25. Huynen M. A., Neumann A. U. Rate of killing of HIV-infected T cells and disease progression. Science. 1996 Jun 28;272(5270):1962–1962. doi: 10.1126/science.272.5270.1962. [DOI] [PubMed] [Google Scholar]
  26. Jurriaans S., Dekker J. T., de Ronde A. HIV-1 viral DNA load in peripheral blood mononuclear cells from seroconverters and long-term infected individuals. AIDS. 1992 Jul;6(7):635–641. doi: 10.1097/00002030-199207000-00004. [DOI] [PubMed] [Google Scholar]
  27. Keet I. P., Krol A., Klein M. R., Veugelers P., de Wit J., Roos M., Koot M., Goudsmit J., Miedema F., Coutinho R. A. Characteristics of long-term asymptomatic infection with human immunodeficiency virus type 1 in men with normal and low CD4+ cell counts. J Infect Dis. 1994 Jun;169(6):1236–1243. doi: 10.1093/infdis/169.6.1236. [DOI] [PubMed] [Google Scholar]
  28. Kim S. Y., Byrn R., Groopman J., Baltimore D. Temporal aspects of DNA and RNA synthesis during human immunodeficiency virus infection: evidence for differential gene expression. J Virol. 1989 Sep;63(9):3708–3713. doi: 10.1128/jvi.63.9.3708-3713.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kirchhoff F., Greenough T. C., Brettler D. B., Sullivan J. L., Desrosiers R. C. Brief report: absence of intact nef sequences in a long-term survivor with nonprogressive HIV-1 infection. N Engl J Med. 1995 Jan 26;332(4):228–232. doi: 10.1056/NEJM199501263320405. [DOI] [PubMed] [Google Scholar]
  30. Klotman M. E., Kim S., Buchbinder A., DeRossi A., Baltimore D., Wong-Staal F. Kinetics of expression of multiply spliced RNA in early human immunodeficiency virus type 1 infection of lymphocytes and monocytes. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):5011–5015. doi: 10.1073/pnas.88.11.5011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Learmont J., Tindall B., Evans L., Cunningham A., Cunningham P., Wells J., Penny R., Kaldor J., Cooper D. A. Long-term symptomless HIV-1 infection in recipients of blood products from a single donor. Lancet. 1992 Oct 10;340(8824):863–867. doi: 10.1016/0140-6736(92)93281-q. [DOI] [PubMed] [Google Scholar]
  32. Menzo S., Bagnarelli P., Giacca M., Manzin A., Varaldo P. E., Clementi M. Absolute quantitation of viremia in human immunodeficiency virus infection by competitive reverse transcription and polymerase chain reaction. J Clin Microbiol. 1992 Jul;30(7):1752–1757. doi: 10.1128/jcm.30.7.1752-1757.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Michael N. L., Chang G., d'Arcy L. A., Ehrenberg P. K., Mariani R., Busch M. P., Birx D. L., Schwartz D. H. Defective accessory genes in a human immunodeficiency virus type 1-infected long-term survivor lacking recoverable virus. J Virol. 1995 Jul;69(7):4228–4236. doi: 10.1128/jvi.69.7.4228-4236.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Michael N. L., Mo T., Merzouki A., O'Shaughnessy M., Oster C., Burke D. S., Redfield R. R., Birx D. L., Cassol S. A. Human immunodeficiency virus type 1 cellular RNA load and splicing patterns predict disease progression in a longitudinally studied cohort. J Virol. 1995 Mar;69(3):1868–1877. doi: 10.1128/jvi.69.3.1868-1877.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Michael N. L., Morrow P., Mosca J., Vahey M., Burke D. S., Redfield R. R. Induction of human immunodeficiency virus type 1 expression in chronically infected cells is associated primarily with a shift in RNA splicing patterns. J Virol. 1991 Mar;65(3):1291–1303. doi: 10.1128/jvi.65.3.1291-1303.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Michael N. L., Vahey M., Burke D. S., Redfield R. R. Viral DNA and mRNA expression correlate with the stage of human immunodeficiency virus (HIV) type 1 infection in humans: evidence for viral replication in all stages of HIV disease. J Virol. 1992 Jan;66(1):310–316. doi: 10.1128/jvi.66.1.310-316.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Montefiori D. C., Pantaleo G., Fink L. M., Zhou J. T., Zhou J. Y., Bilska M., Miralles G. D., Fauci A. S. Neutralizing and infection-enhancing antibody responses to human immunodeficiency virus type 1 in long-term nonprogressors. J Infect Dis. 1996 Jan;173(1):60–67. doi: 10.1093/infdis/173.1.60. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Pantaleo G., Menzo S., Vaccarezza M., Graziosi C., Cohen O. J., Demarest J. F., Montefiori D., Orenstein J. M., Fox C., Schrager L. K. Studies in subjects with long-term nonprogressive human immunodeficiency virus infection. N Engl J Med. 1995 Jan 26;332(4):209–216. doi: 10.1056/NEJM199501263320402. [DOI] [PubMed] [Google Scholar]
  40. Patterson B. K., Till M., Otto P., Goolsby C., Furtado M. R., McBride L. J., Wolinsky S. M. Detection of HIV-1 DNA and messenger RNA in individual cells by PCR-driven in situ hybridization and flow cytometry. Science. 1993 May 14;260(5110):976–979. doi: 10.1126/science.8493534. [DOI] [PubMed] [Google Scholar]
  41. Pelizon C., Diviacco S., Falaschi A., Giacca M. High-resolution mapping of the origin of DNA replication in the hamster dihydrofolate reductase gene domain by competitive PCR. Mol Cell Biol. 1996 Oct;16(10):5358–5364. doi: 10.1128/mcb.16.10.5358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Piatak M., Jr, Saag M. S., Yang L. C., Clark S. J., Kappes J. C., Luk K. C., Hahn B. H., Shaw G. M., Lifson J. D. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259(5102):1749–1754. doi: 10.1126/science.8096089. [DOI] [PubMed] [Google Scholar]
  43. Pomerantz R. J., Seshamma T., Trono D. Efficient replication of human immunodeficiency virus type 1 requires a threshold level of Rev: potential implications for latency. J Virol. 1992 Mar;66(3):1809–1813. doi: 10.1128/jvi.66.3.1809-1813.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Pomerantz R. J., Trono D., Feinberg M. B., Baltimore D. Cells nonproductively infected with HIV-1 exhibit an aberrant pattern of viral RNA expression: a molecular model for latency. Cell. 1990 Jun 29;61(7):1271–1276. doi: 10.1016/0092-8674(90)90691-7. [DOI] [PubMed] [Google Scholar]
  45. Saksela K., Stevens C. E., Rubinstein P., Taylor P. E., Baltimore D. HIV-1 messenger RNA in peripheral blood mononuclear cells as an early marker of risk for progression to AIDS. Ann Intern Med. 1995 Nov 1;123(9):641–648. doi: 10.7326/0003-4819-123-9-199511010-00001. [DOI] [PubMed] [Google Scholar]
  46. Saksela K., Stevens C., Rubinstein P., Baltimore D. Human immunodeficiency virus type 1 mRNA expression in peripheral blood cells predicts disease progression independently of the numbers of CD4+ lymphocytes. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1104–1108. doi: 10.1073/pnas.91.3.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Schnittman S. M., Greenhouse J. J., Lane H. C., Pierce P. F., Fauci A. S. Frequent detection of HIV-1-specific mRNAs in infected individuals suggests ongoing active viral expression in all stages of disease. AIDS Res Hum Retroviruses. 1991 Apr;7(4):361–367. doi: 10.1089/aid.1991.7.361. [DOI] [PubMed] [Google Scholar]
  48. Schnittman S. M., Greenhouse J. J., Psallidopoulos M. C., Baseler M., Salzman N. P., Fauci A. S., Lane H. C. Increasing viral burden in CD4+ T cells from patients with human immunodeficiency virus (HIV) infection reflects rapidly progressive immunosuppression and clinical disease. Ann Intern Med. 1990 Sep 15;113(6):438–443. doi: 10.7326/0003-4819-113-6-438. [DOI] [PubMed] [Google Scholar]
  49. Schwartz D., Sharma U., Busch M., Weinhold K., Matthews T., Lieberman J., Birx D., Farzedagen H., Margolick J., Quinn T. Absence of recoverable infectious virus and unique immune responses in an asymptomatic HIV+ long-term survivor. AIDS Res Hum Retroviruses. 1994 Dec;10(12):1703–1711. doi: 10.1089/aid.1994.10.1703. [DOI] [PubMed] [Google Scholar]
  50. Schwartz S., Felber B. K., Benko D. M., Fenyö E. M., Pavlakis G. N. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):2519–2529. doi: 10.1128/jvi.64.6.2519-2529.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schwartz S., Felber B. K., Fenyö E. M., Pavlakis G. N. Env and Vpu proteins of human immunodeficiency virus type 1 are produced from multiple bicistronic mRNAs. J Virol. 1990 Nov;64(11):5448–5456. doi: 10.1128/jvi.64.11.5448-5456.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Seshamma T., Bagasra O., Trono D., Baltimore D., Pomerantz R. J. Blocked early-stage latency in the peripheral blood cells of certain individuals infected with human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10663–10667. doi: 10.1073/pnas.89.22.10663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sheppard H. W., Lang W., Ascher M. S., Vittinghoff E., Winkelstein W. The characterization of non-progressors: long-term HIV-1 infection with stable CD4+ T-cell levels. AIDS. 1993 Sep;7(9):1159–1166. [PubMed] [Google Scholar]
  54. Simmonds P., Balfe P., Peutherer J. F., Ludlam C. A., Bishop J. O., Brown A. J. Human immunodeficiency virus-infected individuals contain provirus in small numbers of peripheral mononuclear cells and at low copy numbers. J Virol. 1990 Feb;64(2):864–872. doi: 10.1128/jvi.64.2.864-872.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tafuro S., Zentilin L., Falaschi A., Giacca M. Rapid retrovirus titration using competitive polymerase chain reaction. Gene Ther. 1996 Aug;3(8):679–684. [PubMed] [Google Scholar]
  56. Wong M. T., Dolan M. J., Kozlow E., Doe R., Melcher G. P., Burke D. S., Boswell R. N., Vahey M. Patterns of virus burden and T cell phenotype are established early and are correlated with the rate of disease progression in human immunodeficiency virus type 1-infected persons. J Infect Dis. 1996 Apr;173(4):877–887. doi: 10.1093/infdis/173.4.877. [DOI] [PubMed] [Google Scholar]
  57. Zanussi S., Simonelli C., D'Andrea M., Caffau C., Clerici M., Tirelli U., DePaoli P. CD8+ lymphocyte phenotype and cytokine production in long-term non-progressor and in progressor patients with HIV-1 infection. Clin Exp Immunol. 1996 Aug;105(2):220–224. doi: 10.1046/j.1365-2249.1996.d01-746.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zanussi S., Simonelli C., D'Andrea M., Comar M., Bidoli E., Giacca M., Tirelli U., Vaccher E., De Paoli P. The effects of antineoplastic chemotherapy on HIV disease. AIDS Res Hum Retroviruses. 1996 Dec 10;12(18):1703–1707. doi: 10.1089/aid.1996.12.1703. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES