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目的探讨结核分枝杆菌Rv1009结构域多肽的免疫学特性。方法选取33只小白鼠按照融合蛋白免疫情况的不同分为Rv1009组、BCG组和生理盐水组,采用酶联免疫吸附法(ELISA)检测方式对三组小白鼠中的血清特异性抗体滴速情况进行观察。分离小白鼠的脾淋巴细胞后采用体外抗原刺激的方式和MTT比色法对小白鼠脾淋巴细胞增殖指数进行观察。采用ELISA方法检测淋巴细胞悬液中鼠源性γ干扰素(IFN-γ)、白细胞介素-10(IL-10)、白细胞介素-12(IL-12)的产生水平。结果经过研究后发现,结核分枝杆菌Rv1009结构域多肽免疫小白鼠血清特异性抗体的滴速为1:13800,淋巴细胞增殖的指数为(2.42±0.17),明显高于生理盐水组小白鼠的淋巴细胞增值指数(P<0.05)。在ELISA检测方面,结核分枝杆菌Rv1009结构域多肽免疫组小白鼠的IFN-γ、IL-10、IL-12产生水平分别为(1422±31)、(502±10)、(310±12)ng/L,明显高于生理盐水组小白鼠的IFN-γ、IL-10、IL-12产生水平(P<0.05)。结论结核分枝杆菌Rv1009结构域多肽在体外实验研究的过程中效果更好,具有极强的临床研究价值。
Objective To investigate the immunological characteristics of Mycobacterium tuberculosis Rv1009 domain polypeptide. Methods Thirty-three mice were divided into Rv1009 group, BCG group and saline group according to the immunological status of the fusion protein. Serum-specific antibody drip rate in three groups of mice was detected by enzyme linked immunosorbent assay (ELISA) Observe. After splenic lymphocytes were isolated from mice, the splenic lymphocyte proliferation index of mice was observed by MTT colorimetric method in vitro. The production of IFN-γ, IL-10 and IL-12 in lymphocyte suspension was detected by ELISA. Results The results showed that the titer of serum specific antibody against Mycobacterium tuberculosis Rv1009 domain immunized mice was 1: 13800, and the index of lymphocyte proliferation was (2.42 ± 0.17), which was significantly higher than that of mice in saline group Lymphocyte proliferation index (P <0.05). In ELISA, the levels of IFN-γ, IL-10 and IL-12 in mice immunized with Mycobacterium tuberculosis Rv1009 domain polypeptide were (1422 ± 31), (502 ± 10), (310 ± 12) ng / L, which was significantly higher than that of the normal mice (P <0.05). Conclusion Mycobacterium tuberculosis Rv1009 domain polypeptide is more effective in the in vitro experimental research and has strong clinical research value.