(A) Cartoons on the left depict CD28+/+NFκB:GFP+AP1:mCherry+ reporter Jurkat cell cocultured with Raji APCs, with CD80 or CD86 expressed on either Jurkat cell (cis) or Raji cell (trans), ± Abatacept. Flow cytometry histograms show GFP and mCherry expression with the mean fluorescence intensity (MFI) indicated. Histograms in condition 2 are shown as dashed lines in conditions 3–10 as references. Green and red bars summarize GFP (NFκB) and mCherry (AP1) MFI respectively. Bar graph: IL-2 concentration in media after 16 h coculture. Data obtained from ≥5 independent experiments.
(B) Left, CD28, CD80 and CD86 expression on primed mouse T cells. Right, scheme of primed T cells cocultured with anti-CD3ε-loaded splenocytes. Bar graph: % IFNγ+ T cells after 6 h coculture. Data obtained from 3 independent experiments.
(C) Left, CD28, CD80 and CD86 expression on primed human T cells and gating strategy. Right, scheme of primed T cells cocultured with SEB-loaded Raji cell. Bar graph: % IFNγ+ T cells after 6 h coculture. Data obtained from 3 technical replicates using T cells from one donor.
Data shown as mean ± SEM. One-way ANOVA (A) and unpaired two-tailed Student’s t test (B, C): *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See also Figure S2.