Restricted SIV Replication in Rhesus Macaque Lung Tissues During the Acute Phase of Infection
2002; Elsevier BV; Volume: 161; Issue: 3 Linguagem: Inglês
10.1016/s0002-9440(10)64257-5
ISSN1525-2191
AutoresCraig L. Fuller, Yang‐Kyu Choi, Beth A. Fallert, Saverio Capuano, Premeela A. Rajakumar, Michael Murphey‐Corb, Todd A. Reinhart,
Tópico(s)Polyomavirus and related diseases
ResumoThe extent to which simian immunodeficiency virus (SIV) replication in lung tissues contributes to the pool of viruses replicating during acute infection is incompletely understood. To address this issue, in situ hybridization was used to examine SIV replication in multiple lobes of lung from rhesus macaques infected with pathogenic SIV. Despite widespread viral replication in lymphoid and intestinal tissues, the lungs during acute infection harbored rare productively infected cells. Simultaneous immunohistochemical staining for the monocytic marker, CD68, revealed that SIV RNA+ cells in lung tissues during acute infection were CD68−, whereas during AIDS they were predominantly CD68+ and localized in large foci in caudal lobes. SIV RNA+ cells in spleen remained CD68− throughout disease. Since CD68 is also expressed by subpopulations of dendritic cells (DC), we also examined pulmonary CD68+ cells for expression of additional DC markers. DC-LAMP mRNA was abundant in lung tissues and expressed predominantly by CD68− cells, whereas DC-SIGN mRNA was expressed in only very rare cells, indicating that SIV RNA+ cells late in disease were most likely macrophages. These studies of SIV/host interactions demonstrate that macaque lung tissues are minimally infected during acute infection, exhibit changes in predominant target cells for infection, and express very little DC-SIGN. The extent to which simian immunodeficiency virus (SIV) replication in lung tissues contributes to the pool of viruses replicating during acute infection is incompletely understood. To address this issue, in situ hybridization was used to examine SIV replication in multiple lobes of lung from rhesus macaques infected with pathogenic SIV. Despite widespread viral replication in lymphoid and intestinal tissues, the lungs during acute infection harbored rare productively infected cells. Simultaneous immunohistochemical staining for the monocytic marker, CD68, revealed that SIV RNA+ cells in lung tissues during acute infection were CD68−, whereas during AIDS they were predominantly CD68+ and localized in large foci in caudal lobes. SIV RNA+ cells in spleen remained CD68− throughout disease. Since CD68 is also expressed by subpopulations of dendritic cells (DC), we also examined pulmonary CD68+ cells for expression of additional DC markers. DC-LAMP mRNA was abundant in lung tissues and expressed predominantly by CD68− cells, whereas DC-SIGN mRNA was expressed in only very rare cells, indicating that SIV RNA+ cells late in disease were most likely macrophages. These studies of SIV/host interactions demonstrate that macaque lung tissues are minimally infected during acute infection, exhibit changes in predominant target cells for infection, and express very little DC-SIGN. The lungs represent an extremely large interface between the host and environment, with the mucosal/epithelial surface area of an adult human estimated to be 75 m2.1Dunnill MS Postnatal growth of the lung.Thorax. 1967; 17: 329-333Crossref Google Scholar Accordingly, among the collection of outcomes comprising AIDS in human immunodeficiency virus type-1 (HIV-1)-infected individuals, pathology within the lungs is a frequent component. In two independent, retrospective examinations of AIDS autopsy cases spanning the years 1984 to 1999,2Jellinger KA Setinek U Drlicek M Bohm G Steurer A Lintner F Neuropathology and general autopsy findings in AIDS during the last 15 years.Acta Neuropathol (Berl). 2000; 100: 213-220Crossref PubMed Scopus (116) Google Scholar, 3Masliah E DeTeresa RM Mallory ME Hansen LA Changes in pathological findings at autopsy in AIDS cases for the last 15 years.AIDS. 2000; 14: 69-74Crossref PubMed Scopus (254) Google Scholar 75 to 85% of cases demonstrated pathology in lung tissues. The observation that Pneumocystis carinii is one of the most frequent opportunistic pathogens affecting HIV-1-infected individuals also further underscores the importance of the lung as an immunologically impaired environment during HIV-1 infection.2Jellinger KA Setinek U Drlicek M Bohm G Steurer A Lintner F Neuropathology and general autopsy findings in AIDS during the last 15 years.Acta Neuropathol (Berl). 2000; 100: 213-220Crossref PubMed Scopus (116) Google Scholar, 3Masliah E DeTeresa RM Mallory ME Hansen LA Changes in pathological findings at autopsy in AIDS cases for the last 15 years.AIDS. 2000; 14: 69-74Crossref PubMed Scopus (254) Google Scholar, 4Aviram G Fishman JE Sagar M Cavitary lung disease in AIDS: etiologies and correlation with immune status.AIDS Patient Care STDS. 2001; 15: 353-361Crossref PubMed Scopus (34) Google Scholar, 5Graham SM Coulter JB Gilks CF Pulmonary disease in HIV-infected African children.Int J Tuberc Lung Dis. 2001; 5: 12-23PubMed Google Scholar, 6Nathoo KJ Gondo M Gwanzura L Mhlanga BR Mavetera T Mason PR Fatal Pneumocystis carinii pneumonia in HIV-seropositive infants in Harare, Zimbabwe.Trans R Soc Trop Med Hyg. 2001; 95: 37-39Abstract Full Text PDF PubMed Scopus (44) Google Scholar, 7Yparraguirre IT Sant'Anna CC Lopes VG Madi K Necroscopic study of 14 children with AIDS and pneumonia.Rev Assoc Med Bras. 2001; 47: 129-136Crossref PubMed Google Scholar Maintaining appropriate immune surveillance and effector activities in lung tissues, and at the appropriate levels, is important in combating pathogens encountered through pulmonary routes, and is frequently deficient during HIV-1 infection.The extent to which local and systemic HIV-1 replication in humans, and simian immunodeficiency virus (SIV) replication in rhesus macaques, contributes to disruption of immune function in the lungs is not known. HIV-1 DNA can be detected in bronchoalveolar lavages (BAL) obtained from infected individuals throughout the course of disease, but different studies have reported variable frequencies and magnitudes of viral replication.8Koziel H Kim S Reardon C Li X Garland R Pinkston P Kornfeld H Enhanced in vivo human immunodeficiency virus-1 replication in the lungs of human immunodeficiency virus-infected persons with Pneumocystis carinii pneumonia.Am J Respir Crit Care Med. 1999; 160: 2048-2055Crossref PubMed Scopus (51) Google Scholar, 9Lu W Israel-Biet D Virion concentration in bronchoalveolar lavage fluids of HIV-infected patients.Lancet. 1993; 342: 298Abstract PubMed Scopus (15) Google Scholar, 10Nakata K Weiden M Harkin T Ho D Rom WN Low copy number and limited variability of proviral DNA in alveolar macrophages from HIV-1-infected patients: evidence for genetic differences in HIV-1 between lung and blood macrophage populations.Mol Med. 1995; 1: 744-757Crossref PubMed Google Scholar Both HIV-1 and SIV replicate to differing extents throughout the course of infection with higher viral loads detected during the acute phase and terminal stage (AIDS).11Reimann KA Tenner-Racz K Racz P Montefiori DC Yasutomi Y Lin W Ransil BJ Letvin NL Immunopathogenic events in acute infection of rhesus monkeys with simian immunodeficiency virus of macaques.J Virol. 1994; 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Increased levels of viral replication in the lungs appear during the shift from clinical latency to AIDS,17Reinhart TA Rogan MJ Huddleston D Rausch DM Eiden LE Haase AT Simian immunodeficiency virus burden in tissues and cellular compartments during clinical latency and AIDS.J Infect Dis. 1997; 176: 1198-1208Crossref PubMed Scopus (62) Google Scholar although proviral DNA can be detected throughout the entire course of infection.10Nakata K Weiden M Harkin T Ho D Rom WN Low copy number and limited variability of proviral DNA in alveolar macrophages from HIV-1-infected patients: evidence for genetic differences in HIV-1 between lung and blood macrophage populations.Mol Med. 1995; 1: 744-757Crossref PubMed Google Scholar These issues are of importance because, despite the advent of anti-retroviral drug combinations and their demonstrated efficacy in potent suppression of viral replication with associated immune reconstitution,20Bucy RP Hockett RD Derdeyn CA Saag MS Squires K Sillers M Mitsuyasu RT Kilby JM Initial increase in blood CD4(+) lymphocytes after HIV anti-retroviral therapy reflects redistribution from lymphoid tissues.J Clin Invest. 1999; 103: 1391-1398Crossref PubMed Scopus (294) Google Scholar, 21Ho 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: 123-126Crossref PubMed Scopus (3779) Google Scholar, 22Wei X Ghosh SK Taylor ME Johnson VA Emini EA Deutsch P Lifson JD Bonhoeffer S Nowak MA Hahn BH Viral dynamics in human immunodeficiency virus type 1 infection.Nature. 1995; 373: 117-122Crossref PubMed Scopus (2908) Google Scholar there is a persistent viral reservoir that is long-lived and likely replenished.23Finzi D Blankson J Siliciano JD Margolick JB Chadwick K Pierson T Smith K Lisziewicz J Lori F Flexner C Quinn TC Chaisson RE Rosenberg E Walker B Gange S Gallant J Siliciano RF Latent infection of CD4+ T cells provides a mechanism for lifelong persistence of HIV-1, even in patients on effective combination therapy.Nat Med. 1999; 5: 512-517Crossref PubMed Scopus (1741) Google Scholar These viruses may be replicating in cells of the monocyte/macrophage lineage,24Babas T Vieler E Hauer DA Adams RJ Tarwater PM Fox K Clements JE Zink MC Pathogenesis of SIV pneumonia: selective replication of viral genotypes in the lung.Virology. 2001; 287: 371-381Crossref PubMed Scopus (23) Google Scholar, 25Sopper S Demuth M Stahl-Hennig C Hunsmann G Plesker R Coulibaly C Czub S Ceska M Koutsilieri E Riederer P Brinkmann R Katz M ter Meulen V The effect of simian immunodeficiency virus infection in vitro and in vivo on the cytokine production of isolated microglia and peripheral macrophages from rhesus monkey.Virology. 1996; 220: 320-329Crossref PubMed Scopus (57) Google Scholar and it is possible that the lungs serve as such a reservoir.Examination of rhesus macaque lung tissues has demonstrated productive SIV replication predominantly in monocytes/macrophages during AIDS,18Baskin GB Murphey-Corb M Martin LN Soike KF Hu FS Kuebler D Lentivirus-induced pulmonary lesions in rhesus monkeys (Macaca mulatta) infected with simian immunodeficiency virus.Vet Pathol. 1991; 28: 506-513Crossref PubMed Scopus (36) Google Scholar, 19Mankowski JL Carter DL Spelman JP Nealen ML Maughan KR Kirstein LM Didier PJ Adams RJ Murphey-Corb M Zink MC Pathogenesis of simian immunodeficiency virus pneumonia: an immunopathological response to virus.Am J Pathol. 1998; 153: 1123-1130Abstract Full Text Full Text PDF PubMed Scopus (29) Google Scholar although not in all animals that succumb to the immunodeficiency-inducing effects of the virus. During progression to AIDS, the populations of viral variants in approximately 50% of HIV-1-infected individuals switch from a non-syncitium-inducing phenotype (NSI) to syncitium-inducing phenotype (SI) when examined in in vitro culture systems,26Schuitemaker H Koot M Kootstra NA Dereksen MW de Goede RE van Steenwijk RP Lange JM Schattenkerk JK Miedema F Tersmette M Biological phenotype of human immunodeficiency virus type 1 clones at different stages of infection: progression of disease is associated with a shift from monocytotropic to T-cell-tropic virus population.J Virol. 1992; 66: 1354-1360Crossref PubMed Google Scholar, 27Tersmette M Gruters RA de Wolf F de Goede RE Lange JM Schellekens PT Goudsmit J Huisman HG Miedema F Evidence for a role of virulent human immunodeficiency virus (HIV) variants in the pathogenesis of acquired immunodeficiency syndrome: studies on sequential HIV isolates.J Virol. 1989; 63: 2118-2125Crossref PubMed Google Scholar which correlates with a switch in tropism from primary macrophages to established T cell lines. This has led to the supposition that macrophage-tropic (M-tropic) variants replicate early during the course of infection and T cell tropic (T-tropic) variants replicate later. Viral tropism has been based primarily on examination of the replicating properties of viruses in in vitro assays or, in a more limited way, on examination of the genetic and biochemical properties of the envelope glycoproteins of the viruses. Nevertheless, these important analyses do not examine directly the population of cells that actually serve as targets for productive viral replication within host tissues. Recent in situ hybridization (ISH) studies have identified T-lymphocytes as the predominant target cells for productive infection in lymphoid tissues of HIV-1-infected individuals28Schacker T Little S Connick E Gebhard K Zhang ZQ Krieger J Pryor J Havlir D Wong JK Schooley RT Richman D Corey L Haase AT Productive infection of T cells in lymphoid tissues during primary and early human immunodeficiency virus infection.J Infect Dis. 2001; 183: 555-562Crossref PubMed Scopus (105) Google Scholar, 29Zhang Z Schuler T Zupancic M Wietgrefe S Staskus KA Reimann KA Reinhart TA Rogan M Cavert W Miller CJ Veazey RS Notermans D Little S Danner SA Richman DD Havlir D Wong J Jordan HL Schacker TW Racz P Tenner-Racz K Letvin NL Wolinsky S Haase AT Sexual transmission and propagation of SIV and HIV in resting and activated CD4+ T cells.Science. 1999; 286: 1353-1357Crossref PubMed Scopus (730) Google Scholar and SIV-infected rhesus macaques29Zhang Z Schuler T Zupancic M Wietgrefe S Staskus KA Reimann KA Reinhart TA Rogan M Cavert W Miller CJ Veazey RS Notermans D Little S Danner SA Richman DD Havlir D Wong J Jordan HL Schacker TW Racz P Tenner-Racz K Letvin NL Wolinsky S Haase AT Sexual transmission and propagation of SIV and HIV in resting and activated CD4+ T cells.Science. 1999; 286: 1353-1357Crossref PubMed Scopus (730) Google Scholar early during the course of infection. The relative contributions made by T- versus M-tropic variants of SIV to the local pool of productively replicating viruses in the lungs during different stages of infection have not been examined directly in tissues.To determine the extent to which SIV/DeltaB670 productively replicates in lung tissues during acute infection, and in which cell types, we have comprehensively examined lung tissues from adult rhesus macaques during acute infection or AIDS. ISH was used to detect and quantitate SIV viral RNA+ (vRNA+) cells in five lobes of lung from each macaque. Despite widespread viral replication in lymphoid and gastrointestinal tissues, the lung was not a major target organ for productive replication early in the course of infection. As virus replication increased in lung tissues during AIDS, there was a tissue-specific change in the populations of productively infected target cells from rare, predominantly CD68− cells during acute infection to predominantly CD68+ cells during AIDS. Although CD68 has been a widely used marker for monocytes/macrophages, it is also expressed by subpopulations of dendritic cells.30Summers KL Hock BD McKenzie JL Hart DN Phenotypic characterization of five dendritic cell subsets in human tonsils.Am J Pathol. 2001; 159: 285-295Abstract Full Text Full Text PDF PubMed Scopus (165) Google Scholar Therefore, we examined the expression of DC-associated mRNAs and demonstrated that although DC were numerous in parenchymal lung tissue throughout the entire course of disease, they were almost exclusively DC-LAMP+, CD68−, and DC-SIGN− (dendritic-cell-specific ICAM-3 grabbing nonintegrin). DC-SIGN, which can bind HIV and SIV and promote its passage to other susceptible target cells,31Kwon DS Gregorio G Bitton N Hendrickson WA Littman DR DC-SIGN-mediated internalization of HIV is required for trans-enhancement of T cell infection.Immunity. 2002; 16: 135-144Abstract Full Text Full Text PDF PubMed Scopus (441) Google Scholar was expressed in lung tissue only rarely and only late in the disease, whereas it was expressed abundantly in lymphoid and gastrointestinal tissues. These studies provide a comprehensive definition of the virologic events occurring during peak viral replication in SIV-infected macaques within the large pulmonary compartment.Materials and MethodsAnimals and Tissue ProcessingAll animal studies were performed under the approval and guidance of the University of Pittsburgh Institutional Animal Care and Use Committee. The 12 adult rhesus macaques (Macaca mulatta) used in this study were negative for SIV, simian retrovirus (type D), and simian T-lymphotropic viruses -1, -2, and -3 and have been described.17Reinhart TA Rogan MJ Huddleston D Rausch DM Eiden LE Haase AT Simian immunodeficiency virus burden in tissues and cellular compartments during clinical latency and AIDS.J Infect Dis. 1997; 176: 1198-1208Crossref PubMed Scopus (62) Google Scholar Briefly, all animals were inoculated intravenously (I.V.) and sacrificed either 2 weeks PI during the acute phase of infection or on progression to AIDS. During necropsy, transcardial perfusion was performed with 0.9% saline to remove contaminating blood cells from tissues. Tissue specimens were fixed by immersion in fresh 4% paraformaldehyde (Sigma Co., St. Louis, MO)/phosphate-buffered saline (Biowhittaker, Walkersville, MD) (PF/PBS) and processed as described.32Fallert BA Reinhart TA Improved detection of simian immunodeficiency virus RNA by in situ hybridization in fixed tissue sections: combined effects of temperatures for tissue fixation and probe hybridization.J Virol Methods. 2002; 99: 23-32Crossref PubMed Scopus (34) Google Scholar, 33Reinhart TA Fallert BA Pfeifer ME Sanghavi S Capuano III, S Rajakumar P Murphey-Corb M Day R Fuller CL Schaefer TM Increased expression of the inflammatory chemokine CXC chemokine ligand 9/monokine induced by interferon-γ in lymphoid tissues of rhesus macaques during simian immunodeficiency virus infection and acquired immunodeficiency syndrome.Blood. 2002; 99: 3119-3128Crossref PubMed Scopus (69) Google ScholarPlasma Viral LoadsQuantitation of virion-associated RNA in plasma was performed by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) on an ABI Prism 7700 (Applied Biosystems, Foster City, CA) as described.34Fuller DH Rajakumar PA Wilson LA Trichel AM Fuller JT Shipley T Wu MS Weis K Rinaldo CR Haynes JR Murphey-Corb M Induction of mucosal protection against primary, heterologous simian immunodeficiency virus by a DNA vaccine.J Virol. 2002; 76: 3309-3317Crossref PubMed Scopus (104) Google ScholarIn Situ HybridizationISHs were performed as described,17Reinhart TA Rogan MJ Huddleston D Rausch DM Eiden LE Haase AT Simian immunodeficiency virus burden in tissues and cellular compartments during clinical latency and AIDS.J Infect Dis. 1997; 176: 1198-1208Crossref PubMed Scopus (62) Google Scholar, 35Ozden S Aubert C Gonzalez-Dunia D Brahic M Simultaneous in situ detection of two mRNAs in the same cell using riboprobes labeled with biotin and 35S.J Histochem Cytochem. 1990; 38: 917-922Crossref PubMed Scopus (23) Google Scholar, 36Staskus KA Zhong W Gebhard K Herndier B Wang H Renne R Beneke J Pudney J Anderson DJ Ganem D Haase AT Kaposi's sarcoma-associated herpesvirus gene expression in endothelial (spindle) tumor cells.J Virol. 1997; 71: 715-719Crossref PubMed Google Scholar except that overnight hybridizations were performed at 50°C. The riboprobes in these studies encompassed sequences from four regions of the SIVmacBK28 molecular clone34Fuller DH Rajakumar PA Wilson LA Trichel AM Fuller JT Shipley T Wu MS Weis K Rinaldo CR Haynes JR Murphey-Corb M Induction of mucosal protection against primary, heterologous simian immunodeficiency virus by a DNA vaccine.J Virol. 2002; 76: 3309-3317Crossref PubMed Scopus (104) Google Scholar, 37Kornfeld H Riedel N Viglianti GA Hirsch V Mullins JI Cloning of HTLV-4 and its relation to simian and human immunodeficiency viruses.Nature. 1987; 326: 610-613Crossref PubMed Scopus (117) Google Scholar spanning portions of the gag, pol, env, and nef genes, and included positions 47–1130, 1676–3121, 6600–8266, and 8453–9267 (GenBank accession number M19499). Our generation of a rhesus macaque DC-SIGN cDNA (GenBank accession number AF369755) has been described.38Baribaud F Pohlmann S Sparwasser T Kimata MT Choi YK Haggarty BS Ahmad N Macfarlan T Edwards TG Leslie GJ Arnason J Reinhart TA Kimata JT Littman DR Hoxie JA Doms RW Functional and antigenic characterization of human, rhesus macaque, pigtailed macaque, and murine DC-SIGN.J Virol. 2001; 75: 10281-10289Crossref PubMed Scopus (71) Google Scholar A rhesus macaque DC-LAMP cDNA was generated from total RNA extracted from snap-frozen lung using Trizol (Life Technologies, Rockville, MD), and RT-PCR was performed using the Access RT-PCR System (Promega Corporation, Madison, WI) and the following cycling parameters: 48°C for 45 minutes; 94°C for 2 minutes; 30 cycles of 94°C for 30 seconds, 54°C for 30 seconds, and 68°C for 2 minutes; and 68°C for 3 minutes. The primer sequences were YKCDCLAMPF1 (5′-ATGCCCCGGCAGCTCAGCGCGGCGG-3′) and YKCDCLAMPR1 (5′-TTAGATTCTCTGGTATCCAGATGA-3′), based on the human sequences.39Saint-Vis B Vincent J Vandenabeele S Vanbervliet B Pin JJ Ait-Yahia S Patel S Mattei MG Banchereau J Zurawski S Davoust J Caux C Lebecque S A novel lysosome-associated membrane glycoprotein, DC-LAMP, induced upon DC maturation, is transiently expressed in MHC class II compartment.Immunity. 1998; 9: 325-336Abstract Full Text Full Text PDF PubMed Scopus (311) Google Scholar Products were ligated to the pGEM-T vector (Promega) and sequenced completely in both directions using manual and automated strategies (GenBank accession number AF416334).Simultaneous ISH and Immunohistochemistry (IHC)Following stringent ISH and washing, tissue sections were equilibrated in 1X PBS for 5 minutes, and blocked for 1 hour at room temperature in 5% nonfat dry milk/1X PBS supplemented with 1.6% horse serum. Excess blocking agent was removed and the anti-CD68 mAb (clone KP1 (1:50 dilution), Dako Corp., Carpinteria, CA) was applied to the tissue and incubated for 45 minutes at room temperature in a humid chamber. The sections were washed twice in 1X PBS for 3 minutes, and antigen-positive cells were detected by the ABC method (Vector Labs, Burlingame, CA) using 3,3′-diaminobenzidine as the substrate. The reaction was stopped after 8 to 10 minutes by rinsing in 1X PBS, and the tissues were dehydrated in graded ethanols containing 0.3 mol/L ammonium acetate. SIV vRNA+ cells were then detected by emulsion autoradiography with exposure times of 2 to 3 days, and DC-LAMP and DC-SIGN mRNA+ cells were detected with exposure times of 7 days.Image Capture and AnalysisFor the quantitation of the numbers of vRNA+ cells in lungs, 10 random microscopic fields from each lung tissue section were captured through a 60X Plan apochromat objective using the Metaview software package (Universal Imaging Corp., West Chester, PA) and a RT Slider Spot camera (Diagnostic Instruments, Inc., Sterling Heights, MI). Each nucleus and productively infected cell was counted from each captured image using the manual counting feature of Metaview. The percentages of SIV vRNA+ cells that expressed CD68 were examined in the same manner, and 100 SIV vRNA+ cells per section were categorized as CD68 positive or negative. The percentages of DC-associated mRNA+ cells that expressed CD68 were examined in the same manner, and 100 DC-LAMP or DC-SIGN mRNA+ cells per section were categorized as CD68 positive or negative.ResultsMinimal Productive SIV Replication in Lung Tissues during Acute Infection, Despite Widespread Systemic InfectionTo examine comprehensively the viral and immunological events in lung tissues and their timing throughout the course of infection, 12 adult rhesus macaques (Table 1) were inoculated intravenously with a characterized stock of SIV/DeltaB67040Amedee AM Lacour N Gierman JL Martin LN Clements JE Bohm Jr, R Harrison RM Murphey-Corb M Genotypic selection of simian immunodeficiency virus in macaque infants infected transplacentally.J Virol. 1995; 69: 7982-7990PubMed Google Scholar and sacrificed at different times after infection. Of these animals, eight were sacrificed 2 weeks PI, which is when plasma viral loads41Parker RA Regan MM Reimann KA Variability of viral load in plasma of rhesus monkeys inoculated with simian immunodeficiency virus or simian-human immunodeficiency virus: implications for using non-human primate AIDS models to test vaccines and therapeutics.J Virol. 2001; 75: 11234-11238Crossref PubMed Scopus (43) Google Scholar and lymphoid tissue viral loads11Reimann KA Tenner-Racz K Racz P Montefiori DC Yasutomi Y Lin W Ransil BJ Letvin NL Immunopathogenic events in acute infection of rhesus monkeys with simian immunodeficiency virus of macaques.J Virol. 1994; 68: 2362-2370PubMed Google Scholar reach their maximal levels. The remaining four macaques were maintained until they progressed to AIDS, between 21 and 55 weeks PI.Table 1Rhesus Macaques, SIV in Situ Hybridization Signals, and Clinicopathological FindingsSIV vRNA+ cells†SIV vRNA+ cells in spleen, axillary lymph node (AxLN) and jejunum were hybridized with SIV-specific riboprobes: +/−, 100 vRNA+ cells/mm2.AnimalDuration infection*All macaques were inoculated intravenously with a characterized stock of pathogenic isolate SIV/DeltaB670,44 and have been described.37 Clinicopathological data are presented here again for comparative purposes. (wk)SpleenAxLNJejunumSIV vRNA+ cells/nuclei in lung‡Number of vRNA+ cells/tot
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