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目的:Th1与Th2的平衡紊乱是过敏性哮喘的免疫学基础,本研究通过给予小鼠口服免疫不同亲和力屋尘螨抗原Der p2重组耻垢分枝杆菌,来比较其调节免疫应答的差异。方法:1,分别给予6-8周BALB/c小鼠口服接种Mycobacterium Smegmatis(M.S)、pCW-Der p2-rM.S及pCW-Der p2-PEB1-rM.S,在末次免疫后第14、28、56天分别用夹心ELISA法测定小鼠血清及脾脏淋巴细胞培养上清中IFN-γ、IL-2及IL-4的含量。结果:夹心ELISA结果显示,无论是血清还是脾脏淋巴细胞培养上清中,各组IFN-γ、IL-2水平逐渐升高,IL-4水平逐渐降低,8周时更明显,pCW-Der p2-PEB1-rM.S组与M.S、pCW-Der p2-rM.S组相比有明显统计学差异。体外给予Der p2蛋白刺激后,pCW-Der p2-rM.S及pCW-Der p2-PEB1-rM.S组变化更加明显,而M.S组无明显变化。结论:小鼠口服免疫pCW-Der p2-PEB1-rM.S能明显提高Th1型免疫反应,与pCW-Der p2-rM.S组相比有统计学学差异。三种不同亲和力重组耻垢分枝杆菌经口服后均能诱导小鼠产生Th1型免疫应答,且pCW-Der p2-rM.S及pCW-Der p2-PEB1-rM.S诱导产生的Th1优势应答具有Der p2抗原特异性。本研究弥补了粘膜型疫苗低亲和力的不足,为新型口服疫苗的制备和改进创造了条件。
OBJECTIVE: The imbalance of Th1 and Th2 is the immunological basis of allergic asthma. In this study, mice were immunized orally with Der p2 recombinant Mycobacterium smegmatis against different affinity house dust mites to compare their differences in immune response. BALB / c mice were orally inoculated with Mycobacterium Smegmatis (MS), pCW-Der p2-rM.S and pCW-Der p2-PEB1-rM.S respectively at 6-8 weeks after the last immunization. At 28 and 56 days, the levels of IFN-γ, IL-2 and IL-4 in serum and spleen lymphocyte culture supernatant were measured by sandwich ELISA. Results: The results of sandwich ELISA showed that the levels of IFN-γ and IL-2 in serum and splenic lymphocyte culture supernatant increased gradually and the levels of IL-4 gradually decreased in the supernatant of the serum or spleen lymphocytes, and were more obvious at 8 weeks. PCW-Der p2 -PEB1-rM.S group and MS, pCW-Der p2-rM.S group compared to a statistically significant difference. Der p2 protein stimulation in vitro, pCW-Der p2-rM.S and pCW-Der p2-PEB1-rM.S group more obvious changes, while the M.S group had no significant change. Conclusion: The mice immunized with pCW-Der p2-PEB1-rM.S can significantly enhance Th1-type immune response compared with pCW-Der p2-rM.S group. Three different affinity recombinant Mycobacterium smegmatis could induce Th1 type immune response in mice after oral administration, and Th1 predominant response induced by pCW-Der p2-rM.S and pCW-Der p2-PEB1-rM.S Der p2 antigen specificity. This study made up for the lack of low affinity mucosal vaccine, prepared for the new oral vaccine and to create the conditions for improvement.