sCD40L联合TB.HSP70-H22肽刺激树突状细胞诱导特异性抗肿瘤免疫的研究  

Inducement of specific anti-hepatoma immunity of DC activated by HSP70-H22 peptide complexes and sCD40L

作　　者：罗善明[1] 高建[1] 彭明利[2] 

机构地区：[1]重庆医科大学附属第二医院消化内科,重庆400010 [2]重庆医科大学附属第二医院病毒性肝炎研究所,重庆400010

出　　处：《第三军医大学学报》2009年第11期1024-1028,共5页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(2004-54);重庆市卫生局基金(2008-2-214)~~

摘　　要：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 study the specific anti-tumor immunity of dendritic cell （DC） modified by HSP70-tumor peptide complexes and sCD40L in vitro and in vivo. Methods The murine marrow cells were cultured for 7 d, and then randomized to 4 groups： control, only HSP70-H22 peptide complexes, sCD40L, and HSP70-H22 peptide complexes combined with sCD40L. After intervention for 24 h, IL-12 and IL-10 in the supernatant were detected by ELISA assay, CD40 and CD80 of DC by FACS, and the proliferation of spleen lymphocytes by MRL. The ability of spleen lymphocytes activated by DC to H22 cells in vitro was investigated by MTT. The models of murine hepatoma were established, and then randomized to 5 groups （ Ⅰ , Ⅱ ,Ⅲ, Ⅳ and Ⅴ ） , followed by interference with normal saline and DC respectively. At 21 d, mice were sacrificed and the weight of tumor was measured. The levels of IL-10 and IFN-γ in blood serum were detected by ELISA assay. Results Compared with that in the groups of control, only sCD40L, and HSP70-H22 peptide complexes, the level of IL-10 in the group stimulated by HSP70-H22 peptide complexes combined with sCD40L decreased significantly, but other indexes in this group increased significantly （P 〈 0.05 ）. Compared with that in groups Ⅰ ,Ⅱ , Ⅲ and Ⅳ, the level of IL-10 and the tumor weight in group V decreased significantly, but the level of IFN-γ increased significantly （P 〈0.05）. Conclusion Sufficient activation of DC modified by HSP70-tumor peptide complexes and sCD40L can induce powerful specific antitumor immunity, strong killing effect against H22 cells, and efficient inhibition of tumor growth.

关 键 词：热休克蛋白70-H22肽 肝癌 树突状细胞 SCD40L 肿瘤免疫 

分 类 号：R392-33[医药卫生—免疫学] R392.3[医药卫生—基础医学]