CD28、CD40通路共刺激对淋巴细胞增殖反应的影响及CsA的抑制作用

Effects of cyclosporin A on CD28/CTLA4 and CD40/CD40L co-stimulated lymphocyte proliferation

目的 :探讨CD2 8、CD4 0通路共刺激后淋巴细胞增殖反应的变化及免疫抑制剂环孢素A(CsA)、抗CTLA4单克隆抗体对共刺激通路激活后淋巴细胞增殖反应的影响。方法 :采用单克隆抗体与淋巴细胞表面CD3、CD2 8及CD4 0L分子结合产生相应刺激信号 ,根据刺激条件不同设组为 :①抗CD3单抗单刺激 (A组 ) (剂量为 1 0 μg/L、1 0 0μg/L、1 0 0 0 μg/L) ;②抗CD3单抗 +抗CD2 8单抗 (B组 ) ;③抗CD3单抗 +抗CD4 0L单抗 (C组 ) ;④抗CD3单抗 +抗CTLA4单抗 (D组 )共刺激 (A、B、C、D组中抗CD3单抗剂量固定为 1 0 0 μg/L ,其余 3种单抗各设 1 0 μg/L、1 0 0 μg/L、1 0 0 0 μg/L 3种剂量 ) ;⑤抗CD3单抗 +抗CD2 8单抗 +抗CD4 0L单抗 (E组 ) ;⑥抗CD3单抗 +抗CD2 8单抗 +抗CTLA4单抗 (F组 )共刺激 (E、F组中抗CD3单抗和抗CD2 8单抗的剂量固定为 1 0 0 μg/L ,抗CD4 0L单抗和抗CTLA4单抗的剂量各设 1 0 μg/L、1 0 0 μg/L、1 0 0 0 μg/L) ;⑦CsA干预组 :抗CD3单抗 +CsA(G组 ) ;⑧抗CD3单抗 +抗CD2 8单抗 +CsA(H组 ) ;⑨抗CD3单抗 +抗CD2 8单抗 +抗CD4 0L单抗 +CsA(I组 ) ,G、H、I组中所有单抗浓度均固定为1 0 0 μg/L ,CsA浓度设为 1 0 μg/L、1 0 0 μg/L、1 0 0 0 μg/L。采用 [3 H]

Aim: To investigate the effects of cyclosporin A(CsA)and anti CTLA4 monoclonal antibody(mAb) on CD28 and CD40 co stimulated lymphocyte proliferation. Methods:By using human lymphocyte culture system, the lymphocyte proliferation was examined by method of TdR isotope absorption. The experimental groups were divided into CD3 mAb (group A );CD3 mAb+CD28 mAb (group B);CD3 mAb+CD40 L mAb (group C);CD3 mAb+CTLA4 mAb (group D );CD3 mAb+CD28 mAb+CD40L mAb (group E );CD3 mAb+CD28 mAb+CTLA4 mAb (group F);CD3 mAb+CsA (group G );CD3 mAb+CD28 mAb+CsA(group H); CD3 mAb+CD28 mAb+CD40L mAb+CsA (group I). Results:The lymphocyte proliferation in group of anti CD3 mAb+anti CD28 mAb co stimulation was highly increased compared with that in group of anti CD3 mAb stimulation while that was decreased in group of anti CD3 mAb+anti CD40L co stimulation. CsA as well as anti CTLA4 mAb inhibited the lymphocyte proliferation induced by anti CD3 mAb stimulation and anti CD3 mAb+anti CD28 mAb co stimulation. The inhibitory effect of CsA on CD3 mAb stimulated lymphocyte proliferation was more significant. Conclusion: CD28 co stimulatory pathway plays a key role in lymphocyte activation. The stimulatory effects of anti CD40L mAb on lymphocyte proliferation may be concealed by its blockade of interaction between T lymphocyte and antigen presenting cell(APC, including B cells). CsA and CTLA4 mAb can inhibit co stimulated lymphocyte proliferation.