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Role of CD8+ cells in controlling replication of nonpathogenic Simian Immunodeficiency Virus SIVmac1A11
© Van Rompay et al; licensee BioMed Central Ltd. 2006
Received: 17 February 2006
Accepted: 03 April 2006
Published: 03 April 2006
Infection of macaques with the avirulent molecular clone SIVmac1A11 results in transient low viremia and no disease. To investigate if this low viremia is solely due to intrinsic poor replication fitness or is mediated by efficient immune-mediated control, 5 macaques were inoculated intravenously with SIVmac1A11. Three animals that were depleted of CD8+ cells at the start of infection had more prolonged viremia with peak virus levels 1 to 2 logs higher than those of 2 animals that received a non-depleting control antibody. Thus, CD8+ cell-mediated immune responses play an important role in controlling SIVmac1A11 replication during acute viremia.
Simian immunodeficiency virus (SIV) infection of macaques has proven useful for modeling HIV disease pathogenesis and intervention strategies [1–3]. While infection of macaques with most SIV isolates results eventually in an AIDS-like disease, there are also attenuated isolates and clones. SIVmac1A11 is a molecular clone originally derived from a virus isolate from an SIV-infected macaque that was also the source of virulent uncloned SIVmac251 isolates [4, 5]. Although the kinetics are slower than for other isolates, SIVmac1A11 replicates well in vitro and is highly cytopathogenic (with induction of syncytia) in T-cell lines and rhesus macaque peripheral blood mononuclear cells (PBMC); SIVmac1A11 replicates well in macrophage cultures . In early studies, it was observed that SIVmac1A11 inoculation of juvenile macaques resulted in transient viremia and no disease, even after prolonged follow-up for more than 12 years (; unpublished observations). Subsequent studies documented that SIVmac1A11 inoculation of fetal and newborn macaques also resulted in transient viremia and no disease [7, 8]. SIVmac1A11 has a tissue distribution distinct from that of virulent isolates .
Because of these unique properties, SIVmac1A11 has proven useful to study determinants of viral virulence. The genome of SIVmac1A11 has been sequenced, and recombination experiments revealed that differences in more than one region of the viral genome were responsible for the lack of virulence [5, 10]. SIVmac1A11 has also shown promise as a live-attenuated vaccine in both infant and juvenile/adult macaques [10–13].
The transient low-level viremia (peak levels ≤ 4 to 5 log RNA copies per ml plasma) that results from SIVmac1A11 infection suggests either poor intrinsic replication fitness in vivo and/or relatively effective immune control. CD8+ cell depletion experiments (via administration of monoclonal antibody) have demonstrated the important role of CD8+ cell-mediated immune responses in controlling acute and chronic viremia with virulent SIV isolates (such as SIVmac251; ) and chronic viremia with the attenuated clone SIVmac239Δnef ; however, CD8+ cell depletion had no detectable effect on viremia in animals chronically infected with the more attenuated clone SIVmac239Δ3  or with SIVmac1A11 (unpublished data). To our knowledge, no CD8+ cell depletion experiments have been performed during acute infection with nonpathogenic SIV isolates.
Accordingly, we sought to determine the role of CD8+ cell-mediated immune responses on acute SIVmac1A11 viremia. Animals in this study were juvenile rhesus macaques (Macaca mulatta; ~1 year of age), housed in accordance with American Association for Accreditation of Laboratory Animal Care Standards with strict adherence to the "Guide for the Care and Use of Laboratory Animals" . When necessary, the animals were immobilized with ketamine HCL (Parke-Davis, Morris Plains, New Jersey) 10 mg/kg injected intramuscularly.
The 2 control-antibody treated animals had peak plasma viral RNA levels of 4 to 7 × 103 copies/ml at 3 days after virus inoculation (Fig. 1A). For one animal (number 35391), a second smaller peak of viremia was observed on day 17. The levels of viremia in these 2 control animals are thus similar to those described previously for SIVmac1A11-infected juvenile macaques . The 3 CD8+ cell-depleted animals had viral RNA levels during the first 7 days that were indistinguishable from those of the control animals, suggesting that during these early stages, CD8+ cells had no detectable role in controlling SIVmac1A11 replication. However, after an initial decline, viral RNA levels in the CD8+ cell-depleted animals increased from day 10 onwards and reached peak levels of 45,000 to 790,000 on day 17; these values were 1–2 log higher than those of the control animals (p = 0.015, two-tailed t-test comparing day 17 values, and area-under-the curve values for day 0 to 35) and only ~1–2 log lower than peak viremia levels observed with the pathogenic molecular clone SIVmac239 [20–22]. Despite this higher viremia in the CD8+ cell-depleted animals, there were no significant changes in CD4+CD3+ T lymphocyte counts in peripheral blood (Fig. 1D); this study was not designed to monitor CD4+CD3+ T lymphocyte levels in gut-associated lymphoid tissue. Plasma viral RNA levels declined upon the return of CD8+ cells and became undetectable from 28 to 35 days of infection onwards throughout the rest of the observation period (> 6 months). These results indicate that CD8+ cells play a major role in controlling SIVmac1A11 replication because in their absence, peak viremia was higher and the acute viremia phase was significantly prolonged.
Because the cM-T807 antibody depletes both CD8+ T lymphocytes as well as NK cells, the relative contribution of each cell type could not be determined. CD8+ T cells and NK cells inhibit virus replication in vitro through a variety of mechanisms, including cytolytic and non-cytolytic pathways [23–25]. Most NK cells also have the low-affinity Fc-gamma III receptor (CD16), which triggers antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cell-mediated virus inhibition (ADCVI). ADCVI is similar to ADCC but is a measure of virus inhibition, rather than target cell cytotoxicity. ADCVI has been observed ex vivo with serum and effector cells from HIV-infected humans and SIV-infected macaques (Forthal et al, manuscript submitted). Forthal et al. have also demonstrated that ADCVI-mediating antibodies can be found early during HIV-1 infection and reduce HIV-1 yield both by lysis of infected target cells and by the release of beta-chemokines from NK effector cells [27, 28]. In the current study, the SIVmac1A11-infected animals had detectable antiviral IgG antibodies (as measured by whole SIV ELISA techniques; ) at ~2 weeks of infection, and the CD8+ cell-depleted animals had a more rapid increase in antibody titers, possibly due to more antigenic stimulation (Fig. 1E). Early plasma samples were also tested for ADCVI activity; pronounced inhibition (> 70%) was observed in plasma collected at 17 days of infection at a 1:100 dilution in all animals (Fig. 1F). Thus, some of the loss of viremia control following CD8+ cell depletion could be due to the loss of CD8+ NK cells that would likely serve as ADCVI effector cells.
Levels of interleukin-12 and interferon-α were measured in plasma using commercial ELISA-kits (monkey IL-12 ELISA, U-CyTech, Utrecht, the Netherlands; human interferon-α ELISA kit, PBL Biomedical Laboratories, Piscataway, NJ). Although variable levels were detected for both cytokines, there was no correlation with virus levels (data not shown).
In conclusion, this experiment demonstrated that the acute low-level viremia of SIVmac1A11 which is observed following inoculation of untreated animals cannot be explained solely by poor intrinsic replication fitness of the virus; instead, immune responses that are dependent on CD8+ cells limit the magnitude and duration of acute viremia. Viremia of SIVmac1A11 has always been observed to be transient (~2–6 weeks), even following inoculation of fetal and newborn macaques [7, 8]. This indicates that these antiviral immune responses are not abrogated or prevented from emerging during acute SIVmac1A11 viremia and suggests that there is relatively little or no virus-induced immunosuppression. Rather, the anti-SIVmac1A11 immune responses appear able to induce a long-term asymptomatic infection . This is in contrast to infection with virulent SIV isolates, for which irreversible damage to the immune system appears to occur early during the course of infection [9, 30–32]. Accordingly, further experiments that combine avirulent strains such as SIVmac1A11 infection with selective depletions of immune cell populations may prove to be a useful and sensitive model to further unravel precisely the immune responses that are important to control viremia, but that may be difficult to detect during infection with virulent isolates. Attempts to boost or preserve such immune responses may lead to immunotherapeutic strategies that are more effective in achieving long-term control on viremia of virulent virus isolates, including HIV-1.
We thank I. Cazares, T. Dearman, L. Hirst, A. Spinner, W. von Morgenland, the Veterinary Staff, Colony Services, and Clinical Laboratory of the California National Primate Research Center, for expert technical assistance; the Bayer Reference Testing Laboratory (Emeryville, California) for bDNA analysis; K. Reimann (Harvard Medical School, Boston) for assistance and useful discussions. This research was supported by NIH/NIAID grants AI58056 (K.V.R.) and Public Science Health grant RR00169 from the National Center for Research Resources. Reagents used in this work were provided by the NIH Nonhuman Primate Reagent Resource (RR016001 and AI040101) and produced by the National Cell Culture Center.
- Staprans SI, Feinberg MB: The roles of nonhuman primates in the preclinical evaluation of candidate AIDS vaccines. Expert Rev Vaccines 2004,3(4 Suppl):S5-32. 10.1586/14760584.3.4.S5View ArticlePubMedGoogle Scholar
- Haigwood NL: Predictive value of primate models for AIDS. AIDS Reviews 2004,6(4):187-198.PubMedGoogle Scholar
- Van Rompay KKA: Antiretroviral drug studies in non-human primates: a valid animal model for innovative drug efficacy and pathogenesis studies . AIDS Reviews 2005, 7: 67-83.PubMedGoogle Scholar
- Marthas ML, Banapour B, Sutjipto S, Siegel ME, Marx PA, Gardner MB, Pedersen NC, Luciw PA: Rhesus macaques inoculated with molecularly cloned simian immunodeficiency virus. Journal of Medical Primatology 1989, 18: 311-319.PubMedGoogle Scholar
- Luciw PA, Shaw KE, Unger RE, Planelles V, Stout MW, Lackner JE, Pratt-Lowe E, Leung NJ, Banapour B, Marthas ML: Genetic and biological comparisons of pathogenic and nonpathogenic molecular clones of simian immunodeficiency virus (SIVmac). AIDS Res Hum Retroviruses 1992,8(3):395-402.View ArticlePubMedGoogle Scholar
- Banapour B, Marthas ML, Munn RJ, Luciw PA: In vitro macrophage tropism of pathogenic and nonpathogenic molecular clones of simian immunodeficiency virus. Virology 1991, 183: 12-19. 10.1016/0042-6822(91)90113-PView ArticlePubMedGoogle Scholar
- Tarantal AF, Marthas ML, Gargosky SE, Otsyula M, McChesney MB, Miller CJ, Hendrickx AG: Effects of viral virulence on intrauterine growth in SIV-infected fetal rhesus macaques Macaca mulatta. J Acquir Immun Defic Syndr Hum Retrovirol 1995,10(2):129-138. Oct 1View ArticleGoogle Scholar
- Marthas ML, Van Rompay KKA, Otsyula M, Miller CJ, Canfield D, Pedersen NC, McChesney MB: Viral factors determine progression to AIDS in simian immunodeficiency virus-infected newborn rhesus macaques. Journal of Virology 1995,69(7):4198-4205.PubMed CentralPubMedGoogle Scholar
- Lackner AA, Vogel P, Ramos RA, J. D. Kluge JD, Marthas M: Early events in tissues during infection with pathogenic (SIVmac239) and nonpathogenic (SIVmac1A11) molecular clones of simian immunodeficiency virus. American Journal of Pathology 1994,145(2):428-439.PubMed CentralPubMedGoogle Scholar
- Marthas ML, Ramos RA, Lohman BL, Van Rompay KKA, Unger RE, Miller CJ, Banapour B, Pedersen NC, Luciw PA: Viral determinants of simian immunodeficiency virus (SIV) virulence in rhesus macaques assessed by using attenuated and pathogenic molecular clones of SIVmac. J Virol 1993,67(10):6047-6055.PubMed CentralPubMedGoogle Scholar
- Marthas ML, Sutjipto S, Higgins J, Lohman B, Torten J, Luciw PA, Marx PA, Pedersen NC: Immunization with a live, attenuated simian immunodeficiency virus (SIV) prevents early disease but not infection in rhesus macaques challenged with pathogenic SIV. J Virol 1990,64(8):3694-3700.PubMed CentralPubMedGoogle Scholar
- Van Rompay KKA, Greenier JL, Cole KS, Earl P, Moss B, Steckbeck JD, Pahar B, Rourke T, Montelaro RC, Canfield DR, Tarara RP, Miller CJ, McChesney MB, Marthas ML: Immunization of newborn rhesus macaques with simian immunodeficiency virus (SIV) vaccines prolongs survival after oral challenge with virulent SIVmac251. Journal of Virology 2003, 77: 179-190. 10.1128/JVI.77.1.179-190.2003PubMed CentralView ArticlePubMedGoogle Scholar
- Otsyula MG, Miller CJ, Tarantal AF, Marthas ML, Greene TP, Collins JR, Van Rompay KKA, McChesney MB: Fetal or neonatal infection with attenuated simian immunodeficiency virus results in protective immunity against oral challenge with pathogenic SIVmac251. Virology 1996, 222: 275-278. 10.1006/viro.1996.0420View ArticlePubMedGoogle Scholar
- Schmitz JE, Kuroda MJ, Santra S, Sasseville VG, Simon MA, Lifton MA, Racz P, Tenner-Racz K, Dalesandro M, Scallon BJ, Ghrayeb J, Forman MA, Montefiori DC, Rieber EP, Letvin NL, Reimann KA: Control of viremia in simian immunodeficiency virus infection by CD8+ T lymphocytes. Science 1999, 283: 857-860. 10.1126/science.283.5403.857View ArticlePubMedGoogle Scholar
- Metzner KJ, Jin X, Lee FV, Gettie A, Bauer DE, Di Mascio M, Perelson AS, Marx PA, Ho DD, Kostrikis LG, Connor RI: Effects of in vivo CD8+ depletion on virus replication in rhesus macaques immunized with a live, attenuated simian immunodeficiency virus vaccine. Journal of Experimental Medicine 2000,191(11):1921-1931. 10.1084/jem.191.11.1921PubMed CentralView ArticlePubMedGoogle Scholar
- Schmitz JE, Johnson RP, McClure HM, Manson KH, Wyand MS, Kuroda MJ, Lifton MA, Khunkhun RS, McEvers KJ, Gillis J, Piatak M, Lifson JD, Grosschupff G, Racz P, Tenner-Racz K, Rieber EP, Kuus-Reichel K, Gelman RS, Letvin NL, Montefiori DC, Ruprecht RM, Desrosiers RC, Reimann KA: Effect of CD8+ lymphocyte depletion on virus containment after simian immunodeficiency virus SIVmac251 challenge of live attenuated SIVmac239delta3-vaccinated rhesus macaques. J Virol 2005,79(13):8131-8141. 10.1128/JVI.79.13.8131-8141.2005PubMed CentralView ArticlePubMedGoogle Scholar
- National Research Council: Guide for the care and use of laboratory animals. Washington, D. C. , National Academy Press; 1996.Google Scholar
- Van Rompay KKA, Singh RP, Pahar B, Sodora DL, Wingfield C, Lawson JR, Marthas ML, Bischofberger N: CD8+ cell-mediated suppression of virulent simian immunodeficiency virus during tenofovir treatment. Journal of Virology 2004, 78: 5324-5337. 10.1128/JVI.78.10.5324-5337.2004PubMed CentralView ArticlePubMedGoogle Scholar
- Miller CJ, Marthas M, Greenier J, Lu D, Dailey P, Lu Y: In vivo replication capacity rather than in vitro macrophage tropism predicts efficiency of vaginal transmission of simian immunodeficiency virus or simian/human immunodeficiency virus in rhesus macaques. Journal of Virology 1998,72(4):3248-3258.PubMed CentralPubMedGoogle Scholar
- Abel K, Compton L, Rourke T, Montefiori D, Lu D, Rothaeusler K, Fritts L, Bost K, Miller CJ: Simian-human immunodeficiency virus SHIV89.6-induced protection against intravaginal challenge with pathogenic SIVmac239 is independent of the route of immunization and is associated with a combination of cytotoxic T-lymphocyte and alpha interferon responses. Journal of Virology 2003,77(5):3099-3118. 10.1128/JVI.77.5.3099-3118.2003PubMed CentralView ArticlePubMedGoogle Scholar
- Horton H, Vogel TU, Carter DK, Vielhuber K, Fuller DH, Shipley T, Fuller JT, Kunstman KJ, Sutter G, Montefiori DC, Erfle V, Desrosiers RC, Wilson N, Picker LJ, Wolinsky SM, Wang C, Allison DB, Watkins DI: Immunization of rhesus macaques with a DNA prime/modified vaccinia virus Ankara boost regimen induces broad simian immunodeficiency virus (SIV)-specific T-cell responses and reduces initial viral replication but does not prevent disease progression following challenge with pathogenic SIVmac239. J Virol 2002,76(14):7187-7202. 10.1128/JVI.76.14.7187-7202.2002PubMed CentralView ArticlePubMedGoogle Scholar
- Lifson JD, Piatak MJ, Cline AN, Rossio JL, Purcell J, Pandrea I, Bischofberger N, Blanchard J, Veazey RS: Transient early post-inoculation anti-retroviral treatment facilitates controlled infection with sparing of CD4+ T cells in gut-associated lymphoid tissues in SIVmac239-infected rhesus macaques, but not resistance to rechallenge. J Med Primatol 2003,32(4-5):201-210. 10.1034/j.1600-0684.2003.00026.xView ArticlePubMedGoogle Scholar
- Bernstein HB, Kinter AL, Jackson R, Fauci AS: Neonatal natural killer cells produce chemokines and suppress HIV replication in vitro. AIDS Res Hum Retroviruses 2004,20(11):1189-1195.View ArticlePubMedGoogle Scholar
- Oliva A, Kinter AL, Vaccarezza M, Rubbert A, Catanzaro A, Moir S, Monaco J, Ehler L, Mizell S, Jackson R, Li Y, Romano JW, Fauci AS: Natural killer cells from human immunodeficiency virus (HIV)-infected individuals are an important source of CC-chemokines and suppress HIV-1 entry and replication in vitro. Journal of Clinical Investigation 1998,102(1):223-231.PubMed CentralView ArticlePubMedGoogle Scholar
- Dines I, Rumjanek VM, Persechini PM: What is going on with natural killer cells in HIV infection? Int Arch Allergy Immunol 2004,133(4):330-339. 10.1159/000077352View ArticlePubMedGoogle Scholar
- Brenner BG, Gryllis C, Wainberg MA: Role of antibody-dependent cellular cytotoxicity and lymphokine-activated killer cells in AIDS and related diseases. J Leukoc Biol 1991,50(6):628-640.PubMedGoogle Scholar
- Forthal DN, Landucci G, Daar ES: Antibody from patients with acute human immunodeficiency virus (HIV) infection inhibits primary strains of HIV type 1 in the presence of natural killer effector cells. Journal of Virology 2001,75(15):6953-6961. 10.1128/JVI.75.15.6953-6961.2001PubMed CentralView ArticlePubMedGoogle Scholar
- Forthal DN, Landucci G, Phan TB, Becerra J: Interactions between natural killer cells and antibody Fc result in enhanced antibody neutralization of human immunodeficiency virus type 1. J Virol 2005,79(4):2042-2049. 10.1128/JVI.79.4.2042-2049.2005PubMed CentralView ArticlePubMedGoogle Scholar
- Van Rompay KKA, Singh R, Brignolo L, Lawson JR, Schmidt KA, Pahar B, Canfield DR, Tarara RP, Bischofberger N, Marthas M: The clinical benefits of tenofovir for simian immunodeficiency virus-infected macaques are larger than predicted by its effects on standard viral and immunological parameters. Journal of Acquired Immune Deficiency Syndromes 2004,36(4):900-914.View ArticlePubMedGoogle Scholar
- Veazey RS, Lackner AA: Getting to the guts of HIV pathogenesis. J Exp Med 2004,200(6):697-700. 10.1084/jem.20041464PubMed CentralView ArticlePubMedGoogle Scholar
- Li Q, Duan L, Estes JD, Ma ZM, Rourke T, Wang Y, Reilly C, Carlis J, Miller CJ, Haase AT: Peak SIV replication in resting memory CD4+ T cells depletes gut lamina propria CD4+ T cells. Nature 2005,434(7037):1148-1152.PubMedGoogle Scholar
- Mattapallil JJ, Douek DC, Hill B, Nishimura Y, Martin M, Roederer M: Massive infection and loss of memory CD4+ T cells in multiple tissues during acute SIV infection. Nature 2005,434(7037):1093-1097. 10.1038/nature03501View ArticlePubMedGoogle Scholar
- Van Rompay KKA, Marthas ML, Ramos RA, Mandell CP, McGowan EK, Joye SM, Pedersen NC: Simian immunodeficiency virus (SIV) infection of infant rhesus macaques as a model to test antiretroviral drug prophylaxis and therapy: oral 3'-azido-3'-deoxythymidine prevents SIV infection. Antimicrob Agents Chemother 1992,36(11):2381-2386.PubMed CentralView ArticlePubMedGoogle Scholar
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