Fig. 4.
Induction of in vitro T-cell immune responses by depletion of CD15s-expressing T cells. (A) Flow cytometry analysis of CD15s and CD45RA expression by subpopulations of FOXP3+CD4+ T cells from two healthy donors before or after in vitro depletion of CD15s+ cells. (B) CD4+ T-cell responses to NY-ESO-1 peptides after CD15s+ cell depletion. CD15s+ cell-depleted or nondepleted CD4+ T cells isolated from two healthy donors as shown in A were cultured with T-cell–depleted autologous PBMCs pulsed with overlapping NY-ESO-1 peptides covering the entire sequence of NY-ESO-1 protein. IFN-γ–secreting CD4+ T-cell counts were measured by ELISpot assay. (C) CD8+ T-cell responses by the same donors to CMV peptides after CD15s+ cell depletion. PBMCs from HLA-A2+ individuals were CD15s+ cell depleted or nondepleted and cultured in the presence of 10 µM CMV 495–503 HLA-A*0201–restricted peptide for 7 d, and analyzed for the percentages of CMV tetramer-positive CD8+ T cells. Data are representative of two independent experiments.