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目的探讨受sCD40L与结核杆菌HSP70(mycobacterium tuberculosis heat shock protein 70,TB.HSP70)-H22肽联合刺激的树突状细胞(dendritic cell,DC)在体内外的特异性抗肿瘤免疫。方法取小鼠骨髓于培养第7天分为未刺激组、sCD40L组、TB.HSP70-H22肽组、sCD40L+TB.HSP70-H22肽组,共4组,分别给予相应干预,24 h后取上清,ELISA法检测IL-12、IL-10,流式细胞术检测CD40、CD80,混合淋巴细胞反应(MRL)检测淋巴细胞增殖,MTT法检测对肿瘤细胞的杀伤率;建立小鼠肝癌模型,完全随机分为5组(分别给予生理盐水和以上4种干预获得的DC干预),Ⅰ组:生理盐水对照组,Ⅱ组:未刺激DC治疗组,Ⅲ组:sCD40L DC治疗组,Ⅳ组:TB.HSP70-H22肽DC治疗组,Ⅴ组:sCD40L+TB.HSP70-H22肽DC治疗组,于第21天测瘤质量,ELISA法测血清IL-10、IFN-γ。结果sCD40L+TB.HSP70-H22肽组的各项指标与其他各组相比,IL-10显著降低,其他指标均显著升高,差异均有统计学意义(P<0.05)。Ⅴ组与其他各组相比,IL-10和瘤质量均显著降低,IFN-γ显著上升,差异均有统计学意义(P<0.05)。结论受sCD40L与TB.HSP70-H22肽复合物联合刺激后的DC被充分激活,能诱导强有力的特异性抗肿瘤细胞免疫,对H22瘤细胞产生强大的杀伤作用,显著抑制肿瘤的生长。
Objective To investigate the specific antitumor immunity of dendritic cells (DCs) stimulated by sCD40L and mycobacterium tuberculosis heat shock protein 70 (TB.HSP70) -H22 peptide in vitro and in vivo. Methods The bone marrow of mice was divided into 4 groups: unstimulated group, sCD40L group, TB.HSP70-H22 peptide group and sCD40L + TB.HSP70-H22 peptide group on the 7th day. The supernatant was collected and the IL-12 and IL-10 were detected by ELISA. The proliferation of lymphocytes was detected by flow cytometry (CD40), CD80 and mixed lymphocyte reaction (MRL), and the killing rate of tumor cells was detected by MTT assay. , Totally randomly divided into 5 groups (saline intervention and DC intervention respectively). Group I: saline control group, group II: unstimulated DC treatment group, group III: sCD40L DC treatment group, group IV : Group treated with TB.HSP70-H22 peptide DC, GroupⅤ: sCD40L + TB.HSP70-H22 peptide DC treated group, the tumor mass was measured on the 21st day, and the serum IL-10 and IFN-? Results The indexes of sCD40L + TB.HSP70-H22 peptide group were significantly lower than those of other groups, and the other indexes were significantly increased (P <0.05). Compared with other groups, Ⅴ group had significantly lower IL-10 and tumor mass, and IFN-γ significantly increased, with statistical significance (P <0.05). Conclusions DCs stimulated by sCD40L and TB.HSP70-H22 peptide complexes are fully activated, which can induce potent and specific anti-tumor cell immunity, exert a powerful killing effect on H22 tumor cells and significantly inhibit tumor growth.