Macaques were challenged five times weekly by the intrarectal route with 1:10,000 dilution of our SHIV-1157(QNE)Y173H challenge stock (3.67 × 109 copies/ml of viral RNA and 4.88 × 108/ml TCID50 as measured on TZM-bl cells).21 The virus stock was grown from PBMCs and lymph nodes cells of an infected macaque at peak viremia following mitogen stimulation and the stock was titrated in rhesus macaques to select the appropriate challenge dose. Sequence diversity of the viral stock was determined by single-genome amplification.31
SHIV plasma viral RNA measurements were performed at the Immunology Virology Quality Assessment Center Laboratory Shared Resource, Duke Human Vaccine Institute, Durham, NC as described.31
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