Constant mean viral copy number per infected cell in tissues regardless of high, low, or undetectable plasma HIV RNA
R D Hockett, J M Kilby, C A Derdeyn, M S Saag, M Sillers, K Squires, S Chiz, M A Nowak, G M Shaw, R P Bucy, R D Hockett, J M Kilby, C A Derdeyn, M S Saag, M Sillers, K Squires, S Chiz, M A Nowak, G M Shaw, R P Bucy
Abstract
Quantitative analysis of the relationship between virus expression and disease outcome has been critical for understanding HIV-1 pathogenesis. Yet the amount of viral RNA contained within an HIV-expressing cell and the relationship between the number of virus-producing cells and plasma virus load has not been established or reflected in models of viral dynamics. We report here a novel strategy for the coordinated analysis of virus expression in lymph node specimens. The results obtained for patients with a broad range of plasma viral loads before and after antiretroviral therapy reveal a constant mean viral (v)RNA copy number (3.6 log10 copies) per infected cell, regardless of plasma virus load or treatment status. In addition, there was a significant but nonlinear direct correlation between the frequency of vRNA+ lymph node cells and plasma vRNA. As predicted from this relationship, residual cells expressing this same mean copy number are detectable (frequency <2/10(6) cells) in tissues of treated patients who have plasma vRNA levels below the current detectable threshold (<50 copies/ml). These data suggest that fully replication-active cells are responsible for sustaining viremia after initiation of potent antiretroviral therapy and that plasma virus titers correlate, albeit in a nonlinear fashion, with the number of virus-expressing cells in lymphoid tissue.
Figures
References
- Mellors J, Rinaldo C, Gupta P, White RM, Todd JA, Kingsley LA. Prognosis in HIV infection predicted by the quantity of virus in plasma. Science. 1996;272:1167–1170.
- O'Brien W, Hartigan P, Martin D. Changes in plasma HIV-1 RNA and CD4+lymphocyte counts and the risk of progression to AIDS. Rapid and simple PCR assay for quantitation of HIV-1 RNA in plasma: application to acute retroviral infection. N Engl J Med. 1996;334:426–431.
- Carpenter CC, Fischl MA, Hammer SM, Hirsch MS, Jacobsen DM, Katzenstein D, Montaner JS, Richman DD, Saag MS, Schooley RT, et al. Antiretroviral therapy for HIV infection in 1996. Recommendations of an international panel. International AIDS Society-USA. JAMA. 1996;276:146–154.
- Saag MS, Holodniy M, Kuritzkes DR, O'Brien WA, Coombs R, Poscher ME, Jacobsen DM, Shaw GM, Richman DD, Volberding PA. HIV viral load markers in clinical practice. Nat Med. 1996;2:625–629.
- Pantaleo G, Graziosi C, Demarest JF, Butini L, Montroni M, Fox CH, Orenstein JM, Kotler DP, Fauci AS. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease. Nature. 1993;362:355–358.
- Fox CH, Tenner-Racz K, Racz P, Firpo A, Pizzo PA, Fauci AS. Lymphoid germinal centers are reservoirs of human immunodeficiency virus type 1 RNA. J Infect Dis. 1991;164:1051–1057.
- Pantaleo G, Graziosi C, Butini L, Pizzo PA, Schnittman SM, Kotler DP, Fauci AS. Lymphoid organs function as major reservoirs for human immunodeficiency virus. Proc Natl Acad Sci USA. 1991;88:9838–9842.
- Pantaleo G, Graziosi C, Fauci AS. The immunopathogenesis of human immunodeficiency virus infection. N Engl J Med. 1993;328:327–335.
- Embretson J, Zupancic M, Ribas JL, Burke A, Racz P, Tenner-Racz K, Haase AT. Massive covert infection of helper T lymphocytes and macrophages by HIV during the incubation period of AIDS. Nature. 1993;362:359–364.
- Bucy RP, Karr L, Huang GQ, Li J, Devore-Carter D, Honjo K, Lemons A, Murphy KM, Weaver CT. Single-cell analysis of cytokine gene co-expression during naive CD4+T cell phenotype development. Proc Natl Acad Sci USA. 1995;92:7565–7569.
- Karr L, Panoskaltsis-Mortari A, Li J, Devore-Carter D, Weaver CT, Bucy RP. In situhybridization for cytokine mRNA with digoxigenin labeled riboprobes: sensitivity of detection and double label applications. J Immunol Methods. 1995;182:93–106.
- Panoskaltsis-Mortari A, Bucy RP. In situ hybridization with digoxigenin-labeled RNA probes: facts and artifacts. Biotechniques. 1995;18:300–307.
- Bucy RP, Panoskaltsis-Mortari A, Huang GQ, Li J, Karr L, Ross M, Russel JH, Murphy KM, Weaver CT. Heterogeneity of single cell cytokine gene expression in clonal T cell populations. J Exp Med. 1994;180:1251–1262.
- Hockett RD, Janowski KM, Bucy RP. Simultaneous quantitation of multiple cytokine mRNAs by RT-PCR utilizing plate based EIA methodology. J Immunol Methods. 1995;187:273–285.
- Mulder J, McKinney N, Christopherson C, Sninsky J, Greenfield L, Kwok S. Rapid and simple PCR assay for quantitation of HIV-1 RNA in plasma: application to acute retroviral infection. J Clin Microbiol. 1994;32:292–300.
- Cao Y, Ho DD, Todd J, Kokka R, Urdea M, Lifson JD, Piatak M, Jr, Chen S, Hahn BH, Saag MS. Clinical evaluation of branched DNA signal amplification for quantifying HIV type 1 in human plasma. AIDS Res Hum Retroviruses. 1995;11:353–361.
- Haase AT, Henry K, Zupancic M, Sedgewick G, Faust RA, Melroe H, Cavert W, Gebhard K, Staskus K, Zhang Z, et al. Quantitative image analysis of HIV-1 infection in lymphoid tissue. Science. 1996;274:985–989.
- Cavert W, Notermans DW, Staskus K, Wietgrefe SW, Zupancic M, Gebhard K, Henry K, Zhang ZQ, Mills R, McDade H, et al. Kinetics of response in lymphoid tissues to antiretroviral therapy of HIV-1 infection. Science. 1997;276:960–964.
- Orenstein JM, Fox C, Wahl SM. Macrophages as a source of HIV during opportunistic infections. Science. 1997;276:1857–1861.
- Wei X, Ghosh SK, Taylor ME, Johnson VA, Emini EA, Deutsch P, Lifson JD, Bonhoeffer S, Nowak MA, Hahn BH, et al. Viral dynamics in human immunodeficiency virus type 1 infection. Nature. 1995;373:117–122.
- Ho DD, Neumann AU, Perelson AS, Chen W, Leonard JM, Markowitz M. Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1 infection. Nature. 1995;373:123–126.
- Peng H, Reinhart TA, Retzel EF, Staskus KA, Zupancic M, Haase AT. Single cell transcript analysis of human immunodeficiency virus gene expression in the transition from latent to production infection. Virology. 1995;206:16–27.
- Wong JK, Gunthard HF, Havlir DV, Zhang ZQ, Haase AT, Ignacio CC, Kwok S, Emini E, Richman DD. Reduction of HIV-1 in blood and lymph nodes following potent antiretroviral therapy and the virologic correlates of treatment failure. Proc Natl Acad Sci USA. 1997;94:12574–12579.
- Pantaleo G, Cohen OJ, Schacker T, Vaccarezza M, Graziosi C, Rizzardi GP, Kahn J, Fox CH, Schnittman SM, Schwartz DH, et al. Evolutionary pattern of human immunodeficiency virus (HIV) replication and distribution in lymph nodes following primary infection: implications for antiviral therapy. Nat Med. 1998;4:341–345.
- Negulescu PA, Shastri N, Cahalan MD. Intracellular calcium dependence of gene expression in single T lymphocytes. Proc Natl Acad Sci USA. 1994;91:2873–2877.
- Wong JK, Hezareh M, Gunthard HF, Havlir DV, Ignacio CC, Spina CA, Richman DD. Recovery of replication-competent HIV despite prolonged suppression of plasma viremia. Science. 1997;278:1291–1295.
- Finzi D, Hermankova M, Pierson T, Carruth LM, Buck C, Chaisson RE, Quinn TC, Chadwick K, Margolick J, Brookmeyer R, et al. Identification of a reservoir for HIV-1 in patients on highly active antiretroviral therapy. Science. 1997;278:1295–1300.
- Chun TW, Stuyver L, Mizell SB, Ehler LA, Mican JA, Baseler M, Lloyd AL, Nowak MA, Fauci AS. Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. Proc Natl Acad Sci USA. 1997;94:13193–13197.
- Bucy RP. Immune clearance of HIV-1 replication active cells: a model of two patterns of steady state HIV infection. AIDS Res Hum Retroviruses. 1999;15:223–227.
Source: PubMed