结核分枝杆菌Rv1009蛋白的免疫学特性

The immune responses induced by Rv1009 protein of Mycobacterium tuberculosis in mice

目的研究原核表达的结核分枝杆菌Rv1009蛋白在小鼠体内诱导的体液和细胞免疫应答。方法采用皮下包埋的方法,以预先转移到硝酸纤维素膜上的原核表达的Rv1009蛋白免疫小鼠(10只)3次,每次间隔2周。用间接ELISA法检测免疫小鼠血清特异性抗体的滴度。末次免疫完成后4周,处死3只免疫小鼠并分离脾淋巴细胞,体外经PPD(0.2μg/孔)刺激后,用MTT比色法检测免疫小鼠脾淋巴细胞的增殖指数。用ELISA法检测脾淋巴细胞悬液中IFN-γI、L-10及IL-12的水平。结果Rv1009蛋白免疫小鼠血清特异性抗体滴度为1∶1600,淋巴细胞增殖指数为2.46±0.28,IFN-γ含量为1 350±34 pg/ml,IL-10含量为512±15 pg/ml,均显著高于生理盐水对照组;IL-12含量与生理盐水阴性对照组相当,为112±13pg/ml。结论Rv1009有可能作为新型结核疫苗的候选组分。

To evaluate the humoral and cell-mediated immune responses induced by Rv1009 protein of Mycobacterium tuberculosis. C57/BL/6 mice were immunized subcutaneously 3 times at 2 weeks interval on the back of mouse with Rv1008 protein which was transferred to nitrocellulose filter before hand , and the titers of the specific antibodies in sera of the immunized mice were determined by indirect ELISA. Four weeks after the latest immunization , the stimulation indices of the splenic lymphocytes after stimulation with PPD in immunized mice were determined by MTT colorimetry; and the levels of IFN-γ, IL-19 and IL-12 in the supernatants of cultured lymphocytes of immunized mice were assayed by ELISA. It was demonstrated that the titer of the specific antibodies in sera of the immunized mice was rather high, approaching upto 1 ： 1 600, and the stimulation index of the stimulated lymphocytes in the immunized group of mice was higher than that of mice group immunized with saline （2. 464±0.28 versus 0. 894±0.17）. The levels of IFN-γ（1350±34 pg/ml） and IL-10 （512±15 pg/ml） in the cultural supernarants of lymphocyte from the Rv1009 protein- immunized mice were also higher than that of mice immunized with saline （112 13 pg/ml）. However, there was no significant difference between the level of IL-12 in the immunized mice and that of the saline controls. It is evident Rv1009 protein of M. tuberculosis can be used as the candidate component for novel type of tuberculosis vaccine.