Roles of B lymphocyte stimulator and Toll like receptor-4 in systemic lupus erythematosus transgenic mouse model and possible mechanism

HUANG Ning; XIE Wenge; LIU Weifang; LI Wen; WANG Weiping

doi:10.13431/j.cnki.immunol.j.20170132

Abstract：This study was performed to study the roles of B lymphocyte stimulating factor (BLys) and Toll like receptor-4 (TLR-4) in systemic lupus erythematosus (SLE) transgenic mouse model and possible mechanism.A total of 30 cases of SPF EB virus membrane antigen BLLF1 transgenic female mouse were divided randomly into control group,BLys blocking group and TLR-4 blocking group,with 10 mice in each group,at the same time 10 wild type mice were collected as the wild type control group.The mice in BLys blocking group and TLR-4 blocking group were received intraperitoneal injection of anti BR3 monoclonal antibody (500 mg/d) and anti human TLR-4 antibody (250 mg/d) for 10 days,respectively,while the control group and wild type control group were received intraperitoneal injection of equivalent normal saline.Ten days later,caudal vein blood were gathered from all mice to detect TLR-4 mRNA level by RT-PCR method,the levels of interleukin-6 (IL-6),IL-17,tumor necrosis factor alpha (TNF-α) and IL-2 by ELISA,the titer of dsDNA antibody by gold marked immunifaction,the levels of complement 3 (C3),erythrocyte sedimentation rate (ESR) and C-reaction protein (CRP) by conventional biochemical method.HE and Masson staining of kidney was carried out.Data showed that there had no significant difference in BLys in peripheral blood mononuclear cells (PBMC) between wild control group and BLys blocker group,control group and TLR-4 blocker group (P＞0.05).Compared with wild control group and BLys blocker group,BLys in PBMC had increased in the control group and TLR-4 blocker group (P＜0.05),which suggesting that EB virus membrane antigen BLLF1 transgenic mice was SLE model,and BLys blockers can block PBMC BLys expression.There had no significant difference in TLR-4 mRNA,IL-6,IL-17,TNF-α,IL-2,anti-dsDNA antibody,C3,ESR and CRP in wild control group and control group,before and after intervention (P＞0.05).Before intervention,the TLR-4 mRNA,IL-6,IL-17,TNF-α,anti dsDNA antibody,C3,ESR and CRP demonstrated no significantly differences among the control group,BLys blocker group and TLR-4 blockade group (P＞0.05),while after intervention,these indexes were significantly decreased in the two group (P＜0.05),but IL-2 was increased (P＜0.05).Compared with the wild control group,there had differences in TLR-4 mRNA,IL-6,IL-17,IL-2,TNF-α,anti dsDNA antibody,C3,ESR and CRP in other 3 groups (P＜0.05).HE and Masson staining result of BLys blocking group and TLR-4 blocking group were better than that of the control group.Taken together,BLys and TLR-4 may play important roles in SLE,by regulating immune and inflammatory response.