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目的:观察接种以膜蛋白形式表达屋尘螨抗原Der-p2基因的重组BCG(rBCG),对BALB/c小鼠Th细胞免疫应答的影响。方法:以生理盐水为对照,分别将rBCG和BCG经腹腔注射接种于6～8wk龄和新生BALB/c小鼠,用ELISA法测定小鼠血清、脾脏T细胞培养上清(STLCS)中IL4、IFN-γ水平,用双色荧光标记流式细胞仪法测定脾脏细胞Th细胞亚群。结果:接种rBCG和BCG后,成年和新生小鼠血清和STLCS中IL4水平较对照组显著降低、IFN-γ水平显著升高;无论成年鼠还是新生鼠,在CD4+的脾脏细胞中,IFN-γ+细胞的比例明显升高,而IL4+细胞比例明显降低;此外,在两个年龄组小鼠,接种rBCG者脾细胞均产生了较BCG组更高水平的IFNγ;同时,用rBCG免疫的两组小鼠脾脏CD4+IFN-γ+的细胞比例也明显高于BCG免疫各组。结论:无论rBCG还是BCG通过腹腔注射免疫,均可诱导不同年龄BALB/c小鼠产生Th1免疫应答;表达在rBCG细胞壁上的Derp2被当成BCG的成分,为机体免疫系统所识别,抗原再次接触进一步刺激了Derp2特异性的Th1应答。这些结果表明,胞壁型抗原重组BCG可作为有效的疫苗,特异性地调节Th1/Th2平衡。
OBJECTIVE: To observe the effects of recombinant BCG (rBCG) expressing the Der-p2 gene of the house dust mite antigen in membrane protein on the immune response of Th cells in BALB / c mice. Methods: RBCG and BCG were inoculated intraperitoneally into 6-8w of BALB / c mice and neonatal BALB / c mice respectively with saline as control. IL-4 and IL-6 in serum and spleen T cell culture supernatant (STLCS) IFN-γ levels were measured by two-color fluorescence-labeled flow cytometry of Th cells subsets. Results: After inoculation of rBCG and BCG, the level of IL-4 in serum and STLCS of adult and newborn mice was significantly lower than that of control group and the level of IFN-γ was significantly increased. In both adult and neonatal rats, IFN-γ + Cells significantly increased, while the proportion of IL4 + cells was significantly reduced; In addition, in both age groups, spleen cells vaccinated with rBCG produced higher levels of IFNγ than BCG group; meanwhile, two groups immunized with rBCG The proportion of CD4 + IFN-γ + cells in spleen of mice was also significantly higher than that of BCG-immunized groups. Conclusion: Both rBCG and BCG can induce Th1 immune response in BALB / c mice of different ages by intraperitoneal injection. Derp2, which is expressed on the cell wall of rBCG, is recognized as a component of BCG and recognized by the body’s immune system. The antigen re-contacts further Stimulates Derp2-specific Th1 responses. These results indicate that recombinant BCG, a cell wall antigen, can be used as an effective vaccine to specifically regulate Th1 / Th2 balance.