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. 2011 Mar 14;208(3):577–592. doi: 10.1084/jem.20100619

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© 2011 Wang et al.

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Figure 6. — VISTA-Ig inhibited cytokine production by CD4⁺ and CD8⁺ T cells. (A and B) Bulk purified CD4⁺ T cells were stimulated with plate-bound α-CD3 and VISTA-Ig or control-Ig at the stated ratios. Culture supernatants were collected after 24 and 48 h. Levels of IL-2 and IFN-γ were analyzed by ELISA. (C and D) CD4⁺ T cells were sorted into naive (CD25⁻CD44^lowCD62L^hi) and memory (CD25⁻CD44^hiCD62L^low) cell populations. Cells were stimulated with plate-bound α-CD3 in the presence of VISTA-Ig or control-Ig at a ratio of 1:2 (2.5 µg/ml α-CD3 and 5 µg/ml VISTA-Ig or control-Ig). Culture supernatants were collected at 48 h, and the level of IL-2 and IFN-γ was analyzed by ELISA. (E) Bulk purified CD8⁺ T cells were stimulated with plate-bound α-CD3 and VISTA-Ig or control-Ig at the indicated ratios. The level of IFN-γ in the culture supernatant was analyzed by ELISA. For all conditions, supernatant from six duplicated wells was pooled for ELISA analysis and shown as means ± SEM. Shown are representative results from at least three experiments.