vaccination and timing influence siv immune escape viral dynamics in vivo疫苗接种和时间影响体内免疫逃避siv病毒动力学.pdfVIP

Vaccination and Timing Influence SIV Immune Escape Viral Dynamics In Vivo 1 2 1 2 1* Liyen Loh , Janka Petravic , C. Jane Batten , Miles P. Davenport , Stephen J. Kent 1 Department of Microbiology and Immunology, University of Melbourne, Melbourne, Victoria, Australia, 2 Centre for Vascular Research, University of New South Wales, Sydney, New South Wales, Australia CD8þ cytotoxic T lymphocytes (CTL) can be effective at controlling HIV-1 in humans and SIV in macaques, but their utility is partly offset by mutational escape. The kinetics of CTL escape and reversion of escape mutant viruses upon transmission to MHC-mismatched hosts can help us understand CTL-mediated viral control and the fitness cost extracted by immune escape mutation. Traditional methods for following CTL escape and reversion are, however, insensitive to minor viral quasispecies. We developed sensitive quantitative real-time PCR assays to track the viral load of SIV Gag164–172 KP9 wild-type (WT) and escape mutant (EM) variants in pigtail macaques. Rapid outgrowth of EM virus occurs during the first few weeks of infection. However, the rate of escape plateaued soon after, revealing a prolonged persistence of WT viremia not detectable by standard cloning and sequencing methods. The rate of escape of KP9 correlated with levels of vaccine-primed KP9-specific CD8þ T cells present at that time. Similarly, when non-KP9 responder (lacking the restricting Mane-A*10 allele) macaques were infected with SHIVmn229 stock containing a mixture of EM and WT virus, rapid reversion to WT was observed over the first 2 weeks following infection. However, the rate of reversion to WT slowed dramatically over the first month of infection. The serial quantitation of escape mutant viruses ev