Sex-differences in HIV persistence and Immune Dynamics during Reproductive Aging

Dr. Alan Landay’s laboratory at The University of Texas Medical Branch at Galveston will evaluate changes in biological markers of T cell subset immune activation, senescence, exhaustion, maturation, and proliferation in batch by multiparameter flow cytometry. Frozen\thawed PBMC will be evaluated using Aqua Live Dead stain for cell viability and fluorochrome-conjugated monoclonal antibodies to CD3, CD4, CD8, HLA-DR, CD38, CD45RA, CCR7, CD27, CD57, CD28, PD-1, Ki67, and CCR5 for the specific cell populations. Cells will be acquired on an LSRFortessa flow cytometer (BD) using FACS Diva software. Analysis of flow cytometry data will be performed using FlowJo software (Tree Star Inc). Immune activation (CD38+/HLA-DR+), senescence (CD57+CD28-), exhaustion (PD-1+), maturation (CD27/CD45RA), proliferation (Ki-67+), and CCR5 expression will be reported on live singlet gated CD3+\CD4+ or CD3+/CD8+ T cells. Soluble markers of inflammation associated with HIV (TNFa, IL-6, sCD14, sCD163, and CRP) will be evaluated on frozen/thawed aliquots of plasma using commercially available ELISA assays.