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诱导肺局部黏膜免疫对于早期控制结核分枝杆菌(Mycobacterium tuberculosis)十分关键。在我们过去构建的结核基因疫苗pHSP65pep有效诱导脾脏IFN-γ+Th1细胞应答的基础上,为更好诱导呼吸道和肺局部黏膜免疫,采用天然生物多糖壳聚糖(chitosan)作为黏膜递送介质递送结核基因疫苗。制备壳聚糖-pHSP65pep结核黏膜疫苗,滴鼻免疫BALB/c小鼠,检测全身和黏膜T细胞免疫及抗体应答;并以大剂量BCG攻毒,以肺脏HE染色及菌落计数检测免疫保护效果。结果显示,壳聚糖黏膜递送虽不能增加血清HSP65特异性IgG抗体产生,但显著提高了肺灌洗液的SIgA水平;同时显著增强了肺黏膜局部pHSP65pep疫苗诱导的IFN-γ+CD4+Th1细胞应答。攻毒结果证实,壳聚糖递送显著增强了结核基因疫苗的免疫保护效果。表明壳聚糖黏膜递送可显著增强结核基因疫苗诱导的特异性肺黏膜Th1应答和SIgA水平,提高免疫保护效果。提示呼吸道黏膜免疫对于结核保护性免疫具有关键意义。
Induction of local lung mucosal immunity is critical for the early control of Mycobacterium tuberculosis. Based on our previous construction of tuberculosis gene vaccine pHSP65pep to effectively induce spleen IFN-γ + Th1 cell responses, natural biotin chitosan was used as mucosal delivery mediator to deliver nodules for better induction of mucosal immunity in the respiratory tract and lung Gene vaccine. The chitosan-pHH65pep tuberculosis mucosal vaccine was prepared and BALB / c mice were intranasally immunized to detect systemic and mucosal T cell immunity and antibody responses. The mice were challenged with high dose of BCG and the protective effect was evaluated by HE staining and colony counting. The results showed that although chitosan mucosal delivery did not increase serum HSP65-specific IgG antibody production, it significantly increased SIgA level in lung lavage fluid and significantly enhanced pulmonary fibroblast-derived pHSP65pep-induced IFN-γ + CD4 + Th1 cells answer. The challenge results confirmed that the delivery of chitosan significantly enhanced the immunoprotective efficacy of the TB gene vaccine. Chitosan mucosal delivery can significantly enhance the tuberculosis gene vaccine-induced specific pulmonary mucosal Th1 response and SIgA levels, improve immune protection. The results suggest that mucosal immunity in the respiratory tract is of crucial significance for the protective immunity of tuberculosis.