Figure 4. Cirrhotic plasma induces sensitivity to Fas-mediated apoptosis in healthy donor CD27⁺ B-cells.

(A) Purified CD27⁺ and CD27⁻ B-cells from HD were cultured with HD and CIR plasma in the presence and absence of agonistic anti-CD95 mAb (CH11) for 18 hours and then labeled with Annexin-V-FITC and PI. The data represent the frequency of apoptotic cells (Annexin-V⁺) on CD27⁺ and CD27⁻ B-cells with each plasma sample tested. Error bars reflect standard error of the mean. Statistical comparisons done by Wilcoxon test. (B) Impact of blocking anti-Fas mAb, blocking anti-CD40L mAb and two approaches to neutralize circulating Immunoglobulin (Ig) (Fc Blocker and protein A/G-treated CIR plasma) on the expression of Annexin-V on healthy donor CD27⁺ B-cells co-cultured with CIR plasma for 18 hours. (C) Impact of removal and re-introduction of exosomes on healthy donor CD27⁺ B-cells co-cultured with HD or CIR plasma with or without inhibition of Fas for 18 hours. Representative data from three separate experiments with different B-cell donors are shown. (D) Impact of agonist IgG/A/M (pre-cultured for 30 minutes) and/or blocking BAFF mAb on the expression of Annexin-V on healthy donor CD27⁺ B-cells co-cultured with CIR plasma plus agonistic anti-Fas mAb (CH11) for 18 hours. Representative data from three separate experiments with different B-cell donors are shown. Error bars reflect standard error of the mean. Statistical comparisons done by Wilcoxon test.