Figure 1.
Inhibition of HIV-1 in SCT PBMCs. (A) One round of infection with HIV-1-LUC-G virus expressing luciferase. PBMCs were purified from whole blood obtained from individuals with SCT and HbAA control subjects. The cells were treated with phytohemagglutinin and interleukin-2 to activate T cells. PBMCs (12 HbAA, 33% female; 12 HbAS, 83% female) were infected with HIV-1-LUC-G virus expressing luciferase. Luciferase activity was measured at 48 hours’ postinfection. The results are expressed relative to the infection of PBMCs from control subjects, which was set to 100%. Data are shown as mean ± standard deviation. (B-C) HIV-1 (IIIB) replication is repressed in SCT PBMCs. PBMCs were purified and activated as described earlier and infected by HIV-1 (IIIB). Samples were collected at 4 days’ postinfection. Supernatants were collected, and p24 levels were measured by using an enzyme-linked immunosorbent assay (panel B, 8 HbAA, 37.5% female; 6 HbAS, 66.7% female). Cells were collected and RNA was extracted, reverse transcribed, and analyzed with primers for HIV-1 gag gene by real-time PCR using 18S RNA as a reference (panel C, 6 HbAA, 33.3% female; 5 HbAS, 60% female). Data are shown as mean ± standard deviation. (D) Reduction of intracellular HIV-1 p24 in SCT PBMCs. Intracellular p24 levels in PBMCs infected with HIV-1 IIIB were measured by using flow cytometry. A representative histogram shows isotype antibody linked to fluorescein isothiocyanate (FITC-A) staining (black), and p24-FITC-A staining of the infected control PBMCs (blue) and SCT PBMCs (red). Bar graph shows percentage of infected cells ± standard deviation (n = 2 per group, 50% female). (E) Analysis of HIV-1 replication stages in SCT PBMCs. PBMCs purified and activated as described earlier (8 HbAA, 37.5% female; 8 HbAS, 75% female) were infected with HIV-1-LUC-G virus and collected at 14 hours’ postinfection. DNA was extracted and analyzed by real-time PCR on a Roche 4800 using primers for early LTR, late LTR, 2-LTR circles, and integrated viral DNA. β-globin gene was used as a reference. The results show HIV-1 DNA levels in SCT PBMCs relative to control PBMCs.

Inhibition of HIV-1 in SCT PBMCs. (A) One round of infection with HIV-1-LUC-G virus expressing luciferase. PBMCs were purified from whole blood obtained from individuals with SCT and HbAA control subjects. The cells were treated with phytohemagglutinin and interleukin-2 to activate T cells. PBMCs (12 HbAA, 33% female; 12 HbAS, 83% female) were infected with HIV-1-LUC-G virus expressing luciferase. Luciferase activity was measured at 48 hours’ postinfection. The results are expressed relative to the infection of PBMCs from control subjects, which was set to 100%. Data are shown as mean ± standard deviation. (B-C) HIV-1 (IIIB) replication is repressed in SCT PBMCs. PBMCs were purified and activated as described earlier and infected by HIV-1 (IIIB). Samples were collected at 4 days’ postinfection. Supernatants were collected, and p24 levels were measured by using an enzyme-linked immunosorbent assay (panel B, 8 HbAA, 37.5% female; 6 HbAS, 66.7% female). Cells were collected and RNA was extracted, reverse transcribed, and analyzed with primers for HIV-1 gag gene by real-time PCR using 18S RNA as a reference (panel C, 6 HbAA, 33.3% female; 5 HbAS, 60% female). Data are shown as mean ± standard deviation. (D) Reduction of intracellular HIV-1 p24 in SCT PBMCs. Intracellular p24 levels in PBMCs infected with HIV-1 IIIB were measured by using flow cytometry. A representative histogram shows isotype antibody linked to fluorescein isothiocyanate (FITC-A) staining (black), and p24-FITC-A staining of the infected control PBMCs (blue) and SCT PBMCs (red). Bar graph shows percentage of infected cells ± standard deviation (n = 2 per group, 50% female). (E) Analysis of HIV-1 replication stages in SCT PBMCs. PBMCs purified and activated as described earlier (8 HbAA, 37.5% female; 8 HbAS, 75% female) were infected with HIV-1-LUC-G virus and collected at 14 hours’ postinfection. DNA was extracted and analyzed by real-time PCR on a Roche 4800 using primers for early LTR, late LTR, 2-LTR circles, and integrated viral DNA. β-globin gene was used as a reference. The results show HIV-1 DNA levels in SCT PBMCs relative to control PBMCs.

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