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目的比较腺病毒载体(Ad)介导人和小鼠酪氨酸酶相关蛋白2(tyrosinase-relatedpro-tein2,TRP2)修饰小鼠骨髓来源的树突状细胞(BM-DC)诱发抗小鼠黑色素瘤免疫的差异。方法Ad编码人或小鼠TRP2(AdhTRP2或AdmTRP2)体外感染小鼠BM-DC并体内皮下免疫C57BL/6小鼠,7d后取出被免疫小鼠脾细胞行体内细胞毒性T淋巴细胞杀伤试验(invivoCTL)和细胞内IFN-γ染色(ICS)分析CTL的杀伤活性和IFN-γ的产生;或给免疫后小鼠皮下接种小鼠B16.F10黑色素瘤细胞,观察荷瘤小鼠的成活情况。结果invivoCTL和ICS分析显示,AdhTRP2/BM-DC免疫小鼠,其6hCTL杀伤率为(98.7±1.2)%,IFN-γ产生的CD8+T细胞占总CD8+T细胞的(1.25±0.21)%;而AdmTRP2/BM-DC免疫的小鼠,其6hCTL杀伤率和产生IFN-γ的CD8+T细胞比例分别为(28.6±6.3)%和(0.24±0.06)%。荷瘤试验表明,AdhTRP2/BM-DC免疫小鼠后1周皮下接种106B16.F10细胞,观察3个月100%的小鼠无瘤生长;而接种5×104B16.F10细胞至AdmTRP2/BM-DC免疫1周的小鼠,3个月后小鼠成活率仅为40%。结论Ad介导异种(人)TRP2较自身(小鼠)TRP2修饰的BM-DC更为有效地打破肿瘤免疫耐受、诱导强烈的抗黑色素瘤免疫反应,是一种高效的以DC为基础的肿瘤疫苗。
Objective To compare the effects of adenovirus vector (Ad) on mouse bone marrow-derived dendritic cells (BM-DCs) modified by human and mouse tyrosinase-related pro-tein2 (TRP2) Differences in tumor immunity. Methods Ad-CTLs encoding human or mouse TRP2 (AdhTRP2 or AdmTRP2) were used to infect BM-DCs in vitro and C57BL / 6 mice were immunized subcutaneously in vivo. After 7 days, cytotoxic T lymphocyte killing assay (invivoCTL ) And intracellular IFN-γ staining (ICS) CTL cytotoxic activity and IFN-γ production; or immunized mice subcutaneously inoculated with mouse B16.F10 melanoma cells to observe the survival of tumor-bearing mice. Results The results of invivoCTL and ICS showed that the killing rate of AdhTRP2 / BM-DC was (98.7 ± 1.2)% at 6hCTL and (1.25 ± 0.21)% of the total CD8 + T cells by IFN- While the mice immunized with AdmTRP2 / BM-DC had a ratio of 6hCTL killing and IFN-γ producing CD8 + T cells of (28.6 ± 6.3)% and (0.24 ± 0.06)%, respectively. Tumor-bearing experiments showed that mice were immunized subcutaneously with AdhTRP2 / BM-DC 1 week after 106B16.F10 cells were inoculated and 100% of the mice in 3 months were observed tumor-free growth; while 5 × 104B16.F10 cells were inoculated into AdmTRP2 / BM-DC Mice immunized for 1 week had a survival rate of only 40% after 3 months. Conclusion Ad-mediated heterogeneous (human) TRP2 breaks down tumor immune tolerance more efficiently than its own (mouse) TRP2-modified BM-DC and induces a strong anti-melanoma immune response and is an efficient DC-based Tumor vaccine.