重组嵌合毒素Dsg3EC1-2PE40对寻常型天疱疮患者T、B淋巴细胞的影响

目的 探讨重组嵌合毒素Dsg3EC_1-2PE40对寻常型天疱疮患者外周血淋巴细胞的影响。方法 对重组嵌合毒素Dsg3EC_1-2PE40进行表达,并分离纯化;分别以不同浓度重组嵌合毒素作用于寻常型天疱疮患者外周血淋巴细胞,通过ELISPOT法、MTT比色试验及³H-TdR掺入试验来观察重组嵌合毒素对T、B淋巴细胞的影响。结果 纯化后的重组嵌合毒素纯度达80%以上,它能够特异性与天疱疮抗体IgG结合,而与正常人IgG不结合。重组嵌合毒素可使寻常型天疱疮外周血产生抗体的B细胞数量减少约40%左右;在重组嵌合毒素作用下,T细胞的增殖、代谢较重组Dsg3EC_1-2明显降低,两者间差异有统计学意义(P<0.01)。结论 重组嵌合毒素Dsg3EC_1-2PE40可以抑制寻常型天疱疮外周血B细胞的抗体产生作用,对寻常型天疱疮外周血T细胞具有明显的抑制或杀伤作用。

Effects of Recombinant Chimeric Toxin Dsg3EC1-2PE40 on T and B Lymphocytes from Pemphigus Vulgaris

Objective To study the effects of recombinant chimeric toxin Dsg3EC_1-2PE40 on T and B cells from PV patients. Methods The recombinant protein Dsg3EC_1-2PE40 was expressed on BL21TrxB (DE3) cells, then identified and purified. ELISPOT assay was used to detect and quantitate autoantibody-producing B cells in different concentrations of recombinant chimeric toxin, and MTT assay and ³H-TdR assay to observe the metabolism and proliferation of T cells from PV patients in vitro. Results The purity of expressed protein Dsg3EC_1-2PE40 was up to 80%. The number of anti-Dsg3 antibody-producing B cells in PBMC from PV patients decreased by 40% with treatment of Dsg3EC_1-2PE40, which was significantly lower (P<0.01) than those with treatment of Dsg3EC_1-2. Compared with Dsg3EC_1-2, Dsg3EC_1-2PE40 markedly inhibited the metabolism and proliferation of T cells from PV patients (P<0.01). Conclusions Recombinant chimeric toxin Dsg3EC_1-2PE40 can reduce the number of anti-Dsg3 antibody-producing B cells and inhibit or kill T cells from PV patients in vitro.