2 versus 27.9 pg/mL, P = 0.01; Table 2A; Fig. 4A). TNF-β production was strongly correlated with baseline CD27 expression (R2 = 0.34, P = 0.005; Fig. 4B). Interestingly,

strong associations were also observed between baseline CD27 expression and IL-6, IL-12, and TNF-α selleckchem production, although no significant intragroup differences were observed (Fig. 4C,D). Furthermore, cirrhotic B cells also produced less total IgG (but not IgA or IgM) than normal donor B cells (Fig. 4E). Thus, cirrhotic B cells are hyporesponsive to strong activating stimuli, as manifested by impaired up-regulation of CD70, TNF-β, and IgG production. To test the allostimulatory capacity of cirrhotic B cells, relative to HD B cells, we performed a mixed lymphocyte reaction using 48-hour–activated and control B cells to stimulate normal donor CD4+ T cells. Cirrhotic B cells (with or without HCC) were less capable of stimulating alloreactive

CD4+ T-cell proliferation than noncirrhotic HCV patient or HD B cells (Fig. 5A). Interestingly, B-cell allostimulatory capacity was not correlated with memory B-cell frequency, CD86, or HLA-DR (Fig. 5B-E), but did correlate strongly with the degree of up-regulation of CD40 upon activated B cells (R2 = 0.37, P = 0.002; Fig. 5F). B-cell allostimulatory capacity did not significantly correlate with B-cell cytokine production (data not shown). T-cells stimulated by cirrhotic MAPK Inhibitor Library manufacturer B cells were impaired in their capacity to produce TNF-α and TNF-β (Table 2B). In multivariable logistic regression analysis, only CD40 expression and percent CD70+ B cells were the only independent predictors of B-cell allostimulatory capacity (data not shown). Thus, cirrhotic B cells are impaired in their capacity to stimulate CD4+ T cells, an effect that appears to correlate

with impaired up-regulation of costimulation markers after CD40/TLR9 activation. By ELISA, levels of sCD14, a soluble LPS adaptor protein produced and shed by monocytes after LPS exposure,25 were significantly increased in cirrhotic plasma (Fig. 6A). sCD14 concentrations were strongly inversely associated with CD27+ B-cell frequencies (R2 = 0.40, P < 0.001; Fig. 6B). B cells do not express membrane-bound cluster of differentiation 14 (mCD14), but sCD14 上海皓元医药股份有限公司 can directly transfer LPS to myeloid differentiation-2 (MD-2), activating the TLR4 pathway.26 It has also previously been shown that bacterial DNA, a potential TLR9 ligand, can often be detected in cirrhotic plasma.27 We, therefore, investigated the potential role of TLR4 and TLR9 ligands in cirrhotic plasma in activating B cells. HD B cells were cultured with 50% plasma from noncirrhotic (non-CIR; n = 8) or cirrhotic (CIR; n = 8) patients for 72 hours for the measurement of activation (i.e., HLA-DR, CD38, CD27, and CD19). Cirrhotic plasma induced a significant up-regulation of the expression of HLA-DR, up-regulation of CD38, and down-regulation of CD19 (Fig. 6C).