PKC抑制剂Bis Ⅷ对小鼠T淋巴细胞体外活化及增殖的影响

Effect of bisindolylmaleimide Ⅷ on the activation and proliferation of murine T lymphocytes in vitro

研究Bis Ⅷ对小鼠T淋巴细胞体外活化及增殖的影响,并对其免疫调节作用进行初步探讨。分离小鼠淋巴结细胞,制备细胞悬液;以双色荧光抗体染色技术结合流式细胞术检测在ConA的刺激下,Bis Ⅷ对T淋巴细胞早期活化抗原CD69的表达的影响;用羧基荧光素乙酰乙酸(CFDA-SE)染色结合流式细胞术检测在ConA或者PMA和Ion的刺激下,Bis Ⅷ对所分离的T淋巴细胞增殖的影响。终浓度为1μmol/L、3μmol/L、5μmol/L的Bis Ⅷ对ConA刺激的早期活化抗原CD69的表达具有明显的抑制作用(P<0.01);对ConA或者PMA和Ion刺激的T淋巴细胞增殖具有明显的抑制作用(P<0.05),且呈剂量依赖性。Bis Ⅷ对小鼠淋巴细胞的体外活化及增殖均具有明显的抑制作用,是一种潜在的免疫抑制剂。

To investigate the influence of bisindolylmaleimide Ⅷ （Bis Ⅷ） on the activation and proliferation of murine T lymphocytes so as to elucidate the mechanism of its immuno-suppressive effect, cell suspensions were prepared from mouse lymph nodes, the T lymphocytes were treated with different concentrations of Bis Ⅷ and stimulated with polyclonal activator ConA. Meanwhile, fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69. After staining with CFDA-SE, T lymphocytes were stimulated with ConA or phorbol 12-myristate 1,3-acetate （PMA） plus inomycycin （Ion） and flow cytometry was used to assay the proliferation of T lymphocytes. It was demonstrated that Bis Ⅷ in a final concentration of 1, 3 or 5 μmol/L could inhibit significantly the expression of the early activation anti gen CD69 （P〈0.01） and could suppress the proliferative index stimulated by ConA or PMA＋ Ion. It is concluded that Bis Ⅷ can inhibit effectively the activation and proliferation of murine T lymphocytes and it is a potentially immuno depressive agent.