Inhibition of HIV-1 in SCD-derived PBMCs. (A) HPLC analysis of SCD hemoglobin. HPLC of representative blood samples obtained from an SCD subject with HbSS hemoglobin and normal control with HbAA hemoglobin. (B-C) Inhibition of 1 round of HIV-1 infection in SCD-derived PBMCs. PBMCs were purified from whole blood obtained from SCD patients and healthy controls. The cells were activated with PHA and IL-2 and infected with HIV-1-LUC-G virus expressing luciferase. Luciferase activity was measured at 24 hours postinfection (PI) and normalized to the cell numbers. (B) A representative sample that corresponds to panel A. (C) Comparison of 1 round of HIV-1 replication in a cohort of 15 SCD patients. The results are expressed relative to infection of PBMCs from 9 control subjects, which were set to 100%. The means ± standard error (SE) and P value calculated with Student t test are shown. (D) Inhibition of p24 production in SCD-derived PBMCs. PBMCs obtained from 3 SCD patients and 3 controls were activated and infected as described above. Supernatants were collected 72 hours PI, and p24 was measured by ELISA. The means ± SE and P value calculated with Student t test are shown. (E) Inhibition of HIV-1 env expression in SCD-derived PBMCs. PBMCs obtained from 4 SCD patients and 4 controls were activated and infected as described above for 48 hours. RNA was extracted, reverse transcribed, and analyzed with primers for HIV-1 env gene by RT-PCR using 18S RNA as a reference. The means ± SE and P value calculated with Student t test are shown. (F) Inhibition HIV-1 RT in SCD PBMCs. PBMCs obtained from 3 SCD patients and 3 controls were activated with PHA and IL-2 and infected with HIV-1-LUC-G. At 6 hours PI, DNA was extracted and analyzed by RT-PCR on Roche 480 using primers for early LTR and β-globin gene as a reference. The means ± SE are shown (n = 3 for each sample). The means ± SE and P value calculated with Student t test are shown.