Fig 3.

Host antiviral gene expression negatively correlates with ex vivo HIV-1 infectivity in activated PBMC. Individual donor PBMC (n = 43) were assessed for their susceptibility to infection by an CCR5-tropic transmitted/founder strain of subtype B HIV-1 using Tat-regulated Renilla luciferase reporter gene expression to quantify infection. The virus used was an infectious molecular clone that contained the entire ectodomain of HIV-1 WITO4160 and also contained a Renilla luciferase reporter gene. PBMC (n = 43) were activated for 1 day with PHA-P (5 μg/ml), washed, and placed in fresh growth medium without PHA-P. Five-fold serial dilutions of virus were made in quadruplicate for a total of 11 dilutions in 96-well round-bottom tissue-culture plates. PBMC were added and incubated for 6 days. Luminescence was measured using a ViviRen Renilla luciferase kit, and the TCID₅₀ was calculated according to the method described in reference 42. Cell viability was determined by trypan blue staining using a Countess cell counter. Significant negative Spearman correlations were observed between PAF1 and RTF1 (A), TRIM family members (TRIM11 and TRIM26) (B), and BST-2/tetherin (C). P < 0.05 was considered significant. Solid lines depict linear regressions.