Peripheral blood from participants was processed and reserved for
plasma, serum, and peripheral blood mononuclear cells analysis. Soluble factors
were measured independently at each site using commercial enzyme-linked
immunosorbent assay (EIA) kits or multiplexed assays as per
manufacturer’s instructions. Levels of sCD14 and sCD163 were measured by
Quantikine ELISA (R&D Systems, Minneapolis, MN) and the R&D systems
human I-FABP Duo Set EIA kit was used to measure levels of I-FABP. T-cell
activation (percentage of CD4 or CD8 T-cells co-expressing HLA-DR and CD38) was
determined using a whole blood lyse no-wash flow cytometry procedure as
previously described. Commercial antibodies included CD3 PerCP (SK7), CD4 FITC
(SK3), CD8 FITC (SK1), CD38 PE (HB7), and HLA-DR APC (L243) (BD Biosciences, San
Jose, CA). Samples were examined on a BD FACSCalibur or FACS Canto II and data
analyzed using FlowJo version 9.8 (Treestar, Ashland, OR). Cut offs for CD38 and
HLA-DR co-expression were set based on CD Chex Normal control gates. For Ugandan
samples, EIAs were used to additionally quantify serum neopterin
(Immuno-Biological Laboratories America, Minneapolis, MN).
plasma, serum, and peripheral blood mononuclear cells analysis. Soluble factors
were measured independently at each site using commercial enzyme-linked
immunosorbent assay (EIA) kits or multiplexed assays as per
manufacturer’s instructions. Levels of sCD14 and sCD163 were measured by
Quantikine ELISA (R&D Systems, Minneapolis, MN) and the R&D systems
human I-FABP Duo Set EIA kit was used to measure levels of I-FABP. T-cell
activation (percentage of CD4 or CD8 T-cells co-expressing HLA-DR and CD38) was
determined using a whole blood lyse no-wash flow cytometry procedure as
previously described. Commercial antibodies included CD3 PerCP (SK7), CD4 FITC
(SK3), CD8 FITC (SK1), CD38 PE (HB7), and HLA-DR APC (L243) (BD Biosciences, San
Jose, CA). Samples were examined on a BD FACSCalibur or FACS Canto II and data
analyzed using FlowJo version 9.8 (Treestar, Ashland, OR). Cut offs for CD38 and
HLA-DR co-expression were set based on CD Chex Normal control gates. For Ugandan
samples, EIAs were used to additionally quantify serum neopterin
(Immuno-Biological Laboratories America, Minneapolis, MN).