EcoHIV infection of mice establishes latent viral reservoirs in T cells and active viral reservoirs in macrophages that are sufficient for induction of neurocognitive impairment
A. Schematic representation of strategy for assay of integration. In the first round PCR, primers, B1-F and B1-R, from the host B1 repetitive elements and a HIV-specific primer, Fnef, were used to amplify a region of integrated virus and the host flanking region. The second nested QPCR based on the HIV RU5 region was performed using the preamplified products. B. Integration standard curve was generated by logarithmic regression of the amplified IS sample signals. C.-F. Mice (n = 4/system) were pretreated with ABC or RAL 24 h before EcoHIV infection, euthanized 3 days later, and spleens collected for measurement of C. total, D. integrated, and E. 2-LTR circle vDNA; F. spliced vRNA by QPCR of DNA or cDNA as described in Methods.