Suping Wang,
Shanxi Medical University, China
Title: TLR3 signaling pathway on the effect of PBMC recombinant HBsAg immune response in vitro study
Biography
Biography: Suping Wang,
Abstract
Objective: To investigate the effect of on the PBMC recombinant HBsAg immune response in vitro.
Methods: PBMC were divided into 3 groups: blank group, TLR3 activator group (treated with polyI:C) and TLR3 inhibitor group (treated with TLR3 inhibitor), and cultured for 48h. TLR3 signaling pathway molecules in the collected cells were detected by Flow Cytometry, and TLR3/TRIF mRNA and HBV DNA in PBMC by quantitative real-time PCR. Then all the 3 groups were treated with recombinant HBsAg and cultured for 72h, and immune cells were detected by Flow Cytometry.
Results: The percentage of TLR3 (19.21%), pNF-κBp65 (13.73%), pIRF3 positive cells in PBMC (12.64%) among polyI:C group were significantly higher than those in blank group (11.54%, 8.72%, 9.71%, respectively) (P<0.05). The percentage of TLR3 (8.56%) and TRIF (89.61%), NF-κBp65 (89.22%) and IRF3 (91.62%) positive cells in TLR3 inhibitor group were slightly lower than blank group (11.54%, 89.64%, 90.97%, 92.99%, respectively). The expression level of TLR3 (8.98×103) and NF-κBp65 (2.62×104) in polyI:C group were significantly higher than that in blank group (8.09×103 and 2.23×104) (P<0.05). The expression of TLR3 in inhibitor group (7.54×103) was lower than blank group (P=0.045). After PBMC treated with rHBsAg, the proportion of B cells in lymphocytes (5.31%) and plasma cells in B cells (67.71%) among polyI:C group were higher than blank group (4.23% and 58.82%) (P<0.05). The proportions of mDC (4.12%) and pDC (2.96%) in PBMC were also increased compared with blank group (2.51% and 1.73%).
Conclusion: The increased expression of TLR3 protein and activation of NF-κB and IRF3 may promote the maturation of B cells and DC cells, which play an antigen-presenting role and promote formation of plasma cells, and then improve the potential of expression of anti-HBs to enhance the immune response to rHBsAg.