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. Author manuscript; available in PMC: 2009 May 1.

Published in final edited form as: Cell Immunol. 2008 Jul 22;253(1-2):92–101. doi: 10.1016/j.cellimm.2008.05.007

CD25^high PBMC and CD25⁻ PBMC were isolated using a magnetic bead method (as described in Materials and Methods) from peripheral blood of HCs (n=4) and patients with RA (n=6) Representative dot plots of the CD25^high PBMC and CD25⁻ PBMC populations are shown (A). Isolated or combined (1:1) CD25⁻ PBMC and CD25^high PBMC were polyclonally stimulated with plate-bound anti-CD3 and soluble anti-CD28 antibodies for 5 days, and proliferation was measured for the last 18 hr of incubation using [³H]-thymidine incorporation (B). Results are expressed as mean PSL value ± SEM. In co-cultures, CD25^high PBMCs were able to suppress CD25⁻ PBMC proliferation. To assess the suppressive activity of CD25^high PBMC, coculture experiments were performed in which CD25⁻PBMC were cultured in the absence or presence of CD25^high PBMC at 1:1 ratio (C), after which the relative difference in proliferative response between the two conditions was calculated. Results are expressed as mean values ± SEM; NS means not significant.

CD25^high PBMC and CD25⁻ PBMC were isolated using a magnetic bead method (as described in Materials and Methods) from peripheral blood of HCs (n=4) and patients with RA (n=6) Representative dot plots of the CD25^high PBMC and CD25⁻ PBMC populations are shown (A). Isolated or combined (1:1) CD25⁻ PBMC and CD25^high PBMC were polyclonally stimulated with plate-bound anti-CD3 and soluble anti-CD28 antibodies for 5 days, and proliferation was measured for the last 18 hr of incubation using [³H]-thymidine incorporation (B). Results are expressed as mean PSL value ± SEM. In co-cultures, CD25^high PBMCs were able to suppress CD25⁻ PBMC proliferation. To assess the suppressive activity of CD25^high PBMC, coculture experiments were performed in which CD25⁻PBMC were cultured in the absence or presence of CD25^high PBMC at 1:1 ratio (C), after which the relative difference in proliferative response between the two conditions was calculated. Results are expressed as mean values ± SEM; NS means not significant.