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. Author manuscript; available in PMC: 2018 Oct 1.
Published in final edited form as: J Immunol Methods. 2017 Jun 22;449:15–22. doi: 10.1016/j.jim.2017.06.009

Figure 2. Suppression of both CD54 and CD134 Teff surface expression also correlate with suppression of Teff proliferation.

Figure 2

Correlation of percentage suppression of CD25+ Teff (a, day 1; e, day 2; i, day 4) and percentage suppression of CFSE-based Teff proliferation at day 4. Correlation of percentage suppression of CD25+CD54+ Teff (b, day 1; f, day 2; j, day 4) and percentage suppression of CFSE-based Teff proliferation at day 4. Correlation of percentage suppression of CD25+CD134+ Teff (c, day 1; g, day 2; k, day 4) and percentage suppression of CFSE-based Teff proliferation at day 4. The median frequency of CD25+CD134+ on day 2 in Teff cultured without Treg suppression was 25% (range 10–41%). Correlation of percentage suppression of CD25+CD54+CD134+ Teff (d, day 1; h, day 2; l, day 4) and percentage suppression of CFSE-based Teff proliferation at day 4. Spearman’s rank correlation coefficients: a (r=0.772; P<0.0001), b (r=0.866; P<0.0001), c (r=0.900; P<0.0001), d (r=0.953; P<0.0001), e (r=0.925; P<0.0001), f (r=0.915; P<0.0001), g (r=0.943; P<0.0001), h (r=0.955; P<0.0001), i (r=0.962; P<0.0001), j (r=0.961; P<0.0001), k (r=0.952; P<0.0001), and l (r=0.950; P<0.0001). Depending on the number of cells available ratios of 1:2–1:16 Treg:Teff were included for each sample and compared to Teff with anti-CD3/anti-CD28 beads for between 5 to 14 healthy controls from 3 experiments with no replicates. Not all combinations of markers were stained in all assays.