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The dynamics of EIAV viremia and its association with disease cycles have been well characterized, but there has been to date no comprehensive quantitative analyses of the specific tissue sites of EIAV infection and replication in experimentally infected equids during acute disease episodes and during asymptomatic infections in long-term inapparent carriers. To characterize the in vivo site(s) of viral infection and replication, we developed a quantitative competitive PCR assay capable of detecting 10 copies of viral DNA and a quantitative competitive reverse transcription-PCR assay with a sensitivity of about 30 copies of viral singly spliced mRNA. Animals were experimentally infected with one of two reference viruses: the animal-passaged field isolate designated EIAV<jats:sub>Wyo<\/jats:sub>and the virulent cell-adapted strain designated EIAV<jats:sub>PV<\/jats:sub>. Tissues and blood cells were isolated during the initial acute disease or from asymptomatic animals and analyzed for viral DNA and RNA levels by the respective quantitative assays. The results of these experiments demonstrated that the appearance of clinical symptoms in experimentally infected equids coincided with rapid widespread seeding of viral infection and replication in a variety of tissues. During acute disease, the predominant cellular site of viral infection and replication was the spleen, which typically accounted for over 90% of the cellular viral burden. In asymptomatic animals, viral DNA and RNA persisted in virtually all tissues tested, but at extremely low levels, a finding indicative of tight but incomplete immune control of EIAV replication. During all disease states, peripheral blood mononuclear cells (PBMC) were found to harbor less than 1% of the cellular viral burden. These quantitative studies demonstrate that tissues, rather than PBMC, constitute the predominant sites of virus replication during acute disease in infected equids and serve as resilient reservoirs of virus infection, even in the presence of highly effective immune responses that maintain a stringent control of virus replication in long-term inapparent carriers. Thus, these observations with EIAV, a predominantly macrophage-tropic lentivirus, highlight the role of tissues in sequestering lentiviral infections from host immune surveillance.<\/jats:p>","DOI":"10.1128\/jvi.74.7.3112-3121.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:58:35Z","timestamp":1027763915000},"page":"3112-3121","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":71,"title":["Tissue Sites of Persistent Infection and Active Replication of Equine Infectious Anemia Virus during Acute Disease and Asymptomatic Infection in Experimentally Infected Equids"],"prefix":"10.1128","volume":"74","author":[{"given":"Sharon M.","family":"Harrold","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheila J.","family":"Cook","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 405462"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. Frank","family":"Cook","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 405462"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith E.","family":"Rushlow","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles J.","family":"Issel","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky 405462"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald C.","family":"Montelaro","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261,1 and"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.66.12.7328-7335.1992"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.11.7603-7613.1996"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.65.2.1007-1012.1991"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.9.1.100"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.70.11.7842-7850.1996"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(93)90766-V"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/362359a0"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.58.1.67-74.1986"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.20.7086"},{"key":"e_1_3_2_11_2","unstructured":"Genetics Computer Group Program manual for the Wisconsin Package. 1994 Genetics Computer Group Madison Wis"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.71.5.3840-3852.1997"},{"key":"e_1_3_2_13_2","first-page":"272","article-title":"Transmission of equine infectious anemia virus from horses without clinical signs of disease","volume":"180","author":"Issel C. 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