Isolation of naive and memory T cells
Lymphocyte subpopulations were isolated from fresh PBMCs using magnetic
beads (AutoMACS; Miltenyi Biotec) coated with monoclonal Abs to CD4,
CD8, CD45RA, and CD45RO, in accordance with the manufactures’
instructions. The isolated subpopulations were stained with
immunofluorescently-labelled monoclonal Abs to CD4, CD8, CD45RA, and
CD45RO. Isolated CD4+CD45RA+ cells
(naive T cells) and CD4+CD45RO+cells (memory T cells) from both the patients and controls were frozen
in 90% FCS/10% DMSO in the vapor phase oliquid nitrogen until future
use. Samples for the isolation of naive and memory T cells were
available for 24 HC, 21 ART naive, and 18 ART.