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. 2017 Jul;153(1):263–276.e8. doi: 10.1053/j.gastro.2017.03.023

Figure 4.

Figure 4

Effect of circulating soluble mediators on CTLA4 expression. (A) CTLA4 levels in purified CD4+ T conditioned in media supplemented with HC (n = 12) or ALF sera (n = 14) were analyzed using flow cytometry after 24-hour and 48-hour culture. (B) CTL4 levels in purified CD4+ T cells conditioned in media supplemented with sera from ACLF (n = 15), CLD (n = 6) or sepsis (n = 10) sera in comparison to levels induced by culture in the presence of ALF and normal sera. (C) Detection of levels of soluble costimulatory molecules sCD80 and CD86 in sera samples from ALF (n = 20) and HC (n = 10) by enzyme-linked immunosorbent assay (ELISA). (D) sCD80 and sCD86 in sequential samples on days 1 (n = 12), 3 (n = 6), 7 (n = 7), and 14 (n = 5) after admission. (E) Circulating levels of sCD86 and sCD80 in sera from ALF (n = 25), ACLF (n = 20), CLD (n = 15), and sepsis (n = 20) patients determined by ELISA. (F) Healthy and ALF sera were preincubated with anti-CD80 or CD86 to block soluble CD80 or CD86, respectively. Results are representative of 8 independent experiments. Representative histograms gated on CD3+ T cells demonstrating proliferation percentages of carboxyfluorescein succinimidyl esterlabeled CD4+ T cells preconditioned in ALF sera in the absence (left histogram) or presence (right histogram) of pretreatment with anti-CD80 or CD86 blocking antibodies. Proliferation (middle panel) and IL2 secretion (right panel) are from 5 independent experiments.