抗CD3和抗CD28单抗对淋巴细胞产生RANTES的影响

RANTES Produced by Actiated Lymphocytes Costimulated by Anti-CD3 mAb and Anti-CD28 mAb

目的 探讨CD28共刺激通路活化对淋巴细胞产生RANTES的影响与意义,以及免疫抑制剂CsA对RANTES产生的抑制作用。方法 分离健康人外周静脉血中的淋巴细胞。实验分为4组:抗CD3mAb刺激组,抗CD28mAb刺激组,抗CD3mAb+抗CD28mAb共刺激组(CD28共刺激组),未加任何刺激作为对照组。PDTC和CsA以不同终浓度干预CD28共刺激组。采用ELISA方法测定培养上清中RANTES含量,流式细胞术检测淋巴细胞CD25和CCR5表达率。结果 培养48h后,CD28共刺激组RANTES产生显著增加(P<0.05);不同剂量PDTC和CsA对CD28共刺激后淋巴细胞产生RANTES均具有抑制作用;CD28共刺激后淋巴细胞CD25表达率显著增高(P<0.05),而CCR5表达率显著降低(P<0.05)。结论 CD28通路共刺激后淋巴细胞产生RNATES显著增加,这一效应受到NF-κB信号通路的调控。淋巴细胞产生RANTES及其对RANTES的反应均受到CD28共刺激信号调控。抑制淋巴细胞产生RANTES可能是CsA发挥作用的重要分子免疫学机制。

Purpose: To investigate the changes of regulated upon activation, normal T cell expressed and secreted (RANTES) level produced by activated lymphocytes costimulated by CD28 pathway and the effects of Cyclosporin A(CsA). Methods: Human peripheral lymphocytes separated by Ficoll were divided into four groups: anti-CD3mAb group, anti-CD28mAb group, CD28 costimulation group and control group. CD28 costimulation group was treated by different concentrations of PDTC or CsA. RANTES levels were detected by ELISA and CD25 and CCR5 expression of lymphocytes were detected by FCM. Results: RANTES concentration was significantly higher in CD28 costimulation group than other group (P < 0.05). RANTES secreted by lymphocytes could be inhibited by PDTC or CsA. CD25 expression was upregulated after lymphocytes costimulated by CD28 pathway, on the contrary, CCR5 was significantly downregulated (P < 0.05). Conclusions: CD28 costimulating lymphocytes play a important role on RANTES generation which regulated by NF-κB pathway. Lymphocytes producing RANTES and their reaction to RANTES were both related to CD28 pathway. Inhibiting the secretion of RANTES may be one of the important immunosuppressive mechanisms of CsA.