结核杆菌H37Ra免疫小鼠后脾淋巴细胞的杀伤活性及免疫机制的研究医学论文.docVIP

结核杆菌H37Ra免疫小鼠后脾淋巴细胞的杀伤活 性及免疫机制的研究 ：王利娴，卢贤瑜，李惠，阳强 关键词】结核杆菌H37Ra;，Ag85B蛋白;，小鼠淋巴细胞;，穿孔 素;，颗粒酶B Study on the cytotoxicity of spleen lymphocytes and immune mechanisms in mice immunized by Mycobacterium tuberculosis H37Ra [Abstract] AIM: To explore the specific cytotoxicity of spleen lymphocytes and the immune mechanisms in mice immunized by Mycobacterium tuberculosis (MTB) H37Ra. METHODS: At the various interval (30days, 60days) after the mice were immunized by MTB H37Ra，BCG and PBS, the spleen lymphocytes of the immunized mice were used as the effector cells while the Sp2/0 cells expressing the secreted protein Ag85B were used as the target cells, and the cytotoxicity of spleen lymphocytes in the immunized mice was measured by MTT assay. Spleen lymphocytes were collected at 60 days after immunization and stimulated with PPD，and the expression of perforin, granzyme B on mRNA level was detected by RTPCR. RESULTS: The cytotoxicity of spleen lymphocytes in the group immunized by MTB H37Ra was significantly higher than that of PBS control group(P 005)，and slightly higher than that of BCG group. The mRNA expression of perforin, granzyme B in H37Ra group and BCG group was significantly higher than that in PBS control group (P 005); the expression of perforin mRNA in H37Ra group was significantly higher than that in BCG group (P 005)，but there was no obvious difference with regard to the expression of granzyme B mRNA between H37Ra group and BCG group. CONCLUSION: The cytotoxicity of spleen lymphocytes is enhanced after mice were immunized by MTB H37Ra，which may be related to the expression of perforin and granzyme B. [Keywords]MTB H37Ra; Ag85B protein; spleen lymphocyte; perforin; granzyme B ［ ］目的：探讨结核杆菌H37Ra免疫小鼠后，产生特异性的 脾淋巴细胞杀伤活性及其免疫机制。方法：（1）分别以结核杆菌 H37Ra、BCG和PBS免疫小鼠后第30天及第60天，分离各组 小鼠脾淋巴细胞作为效应细胞，以表达Ag85B分泌蛋白的Sp2/0 细胞为靶细胞，用MTT比色法检测免疫小鼠脾淋巴细胞的杀伤率; ⑵在免疫后第60天，小鼠脾淋巴细胞经PPD刺激后，以RTPCR法 检测穿孔素mRNA及颗粒酶B mRNA的表达。结果：（1）结核杆菌 H37Ra组脾淋巴细胞的杀伤率明显高于PBS对照组（P 0.05），略高 于BCG组；（2）H37Ra组和BCG组穿孔素、颗粒酶m