HIV-RNA in blood plasma was quantified locally at all study-visits, by real-time PCR (Roche or Abbott) as described above for the semen.
Blood samples were centralized for total cell-associated HIV-DNA quantification. Thawed whole blood was analyzed with the same ultrasensitive real-time PCR method as described above for the semen (Generic HIV-DNA assay, Biocentric, Bandol, France) [20 (link)]. Each entire DNA extract (quantified with Nanodrop as previously described) was tested in two replicates, and the results were reported as the number of HIV-DNA copies per 106 PBMC, taking into account the whole blood cell count.
Frozen blood plasma samples from the biobank were addressed to the Paris Pasteur Institute and Inserm U1184. Levels of IL-6, IP-10, sCD14 and sCD163 were measured in duplicate with specific ELISA assays (Human IL-6 Platinum ELISA, eBioscience; Human quantikine CXCL10 ELISA, R&D ELISA R&D; Human CD14 DuoSet ELISA and Human CD163 DuoSet ELISA, R&D Systems, Minneapolis, Minnesota). Samples with undetectable levels of a given analyte were arbitrarily attributed half the minimal detectable value.
Blood samples were centralized for total cell-associated HIV-DNA quantification. Thawed whole blood was analyzed with the same ultrasensitive real-time PCR method as described above for the semen (Generic HIV-DNA assay, Biocentric, Bandol, France) [20 (link)]. Each entire DNA extract (quantified with Nanodrop as previously described) was tested in two replicates, and the results were reported as the number of HIV-DNA copies per 106 PBMC, taking into account the whole blood cell count.
Frozen blood plasma samples from the biobank were addressed to the Paris Pasteur Institute and Inserm U1184. Levels of IL-6, IP-10, sCD14 and sCD163 were measured in duplicate with specific ELISA assays (Human IL-6 Platinum ELISA, eBioscience; Human quantikine CXCL10 ELISA, R&D ELISA R&D; Human CD14 DuoSet ELISA and Human CD163 DuoSet ELISA, R&D Systems, Minneapolis, Minnesota). Samples with undetectable levels of a given analyte were arbitrarily attributed half the minimal detectable value.