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目的研究重组白假丝酵母菌(Candida albicans)天冬氨酸蛋白酶2(Sap2)蛋白与致敏树突状细胞(DC)联合免疫对系统性白假丝酵母菌感染小鼠的保护效果。方法 PCR克隆白假丝酵母菌Sap2,与pMD19-T-Sap2载体转化至DH5α,阳性质粒与pET32a构建表达载体,转化至BL21(DE3),诱导目的蛋白表达。将白假丝酵母菌孢子、菌丝混悬液致敏小鼠DC后,3组小鼠分别免疫接种重组蛋白与致敏DC、重组Sap2蛋白、致敏DC,每组接种3次。再造系统性白假丝酵母菌感染模型,观察小鼠生存期、肾病理切片、观察保护性。结果成功构建pET32a-Sap2原核表达载体,目的蛋白被诱导表达并收集纯化。DC、重组Sap2蛋白、重组Sap2蛋白与DC均能有效延长系统性白假丝酵母菌感染小鼠生存期、保护小鼠肾脏,其中重组蛋白与DC联合免疫较单一的DC或Sap2免疫效果更佳。结论重组Sap2蛋白联合DC是系统性白假丝酵母菌感染重组疫苗的较理想免疫组合。
Objective To study the protective effect of combined immunization with Candida albicans aspartic proteinase 2 (Sap2) and sensitized dendritic cells (DCs) on systemic Candida albicans infection in mice. Methods PCR was used to clone Candida albicans Sap2, transformed into pD32a with pMD19-T-Sap2 vector and constructed expression vector pET32a. The recombinant plasmid was transformed into BL21 (DE3) to induce the expression of the target protein. After the C. albicans spores and mycelium suspension were sensitized to DC in mice, the three groups of mice were immunized with the recombinant proteins and sensitized DC, recombinant Sap2 protein, and sensitized DC, respectively, three times in each group. The model of systemic Candida albicans infection was reconstructed to observe the survival period of mice and pathological sections of kidney to observe the protective effect. Results The prokaryotic expression vector pET32a-Sap2 was successfully constructed. The target protein was induced to express and purified. DC, recombinant Sap2 protein, recombinant Sap2 protein and DC can effectively prolong the survival of mice infected with systemic Candida albicans and protect mice kidney, and the combined immunization with recombinant protein and DC is more effective than single DC or Sap2 . Conclusion Recombinant Sap2 protein combined with DC is the ideal immune combination of systemic Candida albicans infection with recombinant vaccine.