Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids

Equine infectious anemia virus (EIAV) infection of horses is characterized by recurring cycles of disease and viremia that typically progress to an in apparent infection in which clinical symptoms are absent as host immune res ponses maintain control of virus replication indefinitely. The dynamics of EIAV viremia and its association with disease cycles have been well charact erized, but there has been to date no comprehensive quantitative analyses o f the specific tissue sites of EIAV infection and replication in experiment ally infected equids during acute disease episodes and during asymptomatic infections in long-term inapparent carriers. To characterize the in vivo si te(s) of viral infection and replication, we developed a quantitative compe titive PCR assay capable of detecting LO copies of viral DNA and a quantita tive competitive reverse transcription-PCR assay with a sensitivity of abou t 30 copies of viral singly spliced mRNA. Animals were experimentally infec ted with one of two reference viruses: the animal-passaged held isolate des ignated EIAV(Wyo) and the virulent cell-adapted strain designated EIAV(PV). 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 demonstrat ed that the appearance of clinical symptoms in experimentally infected equi ds coincided with rapid widespread seeding of viral infection and replicati on in a variety of tissues. During acute disease, the predominant cellular site of viral infection and replication was the spleen, which typically acc ounted for over 90% of the cellular viral burden. In asymptomatic animals, viral DNA and RNA persisted in virtually all tissues tested, but at extreme ly low levels, a finding indicative of tight but incomplete immune control of EIAV replication. During all disease states, peripheral blood mononuclea r cells (PBMC) were found to harbor less than 1% of the cellular viral burd en. 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, ev en in the presence of highly effective immune responses that maintain a str ingent 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.