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目的探讨腺病毒载体(Ad)介导异种黑色素瘤相关抗原酪氨酸酶相关蛋白2(TRP2)诱发抗肿瘤免疫与自身免疫性损伤的关系。方法用Ad编码的人或小鼠TRP2(AdhTRP2或AdmTRP2)皮内免疫C57BL/6小鼠,将小鼠分为正常对照组(d1703)、AdhTRP2和AdmTRP2免疫组。体内细胞毒性T淋巴细胞(CTL)杀伤试验和细胞内干扰素γ(IFNγ)染色(ICS)分析CTL杀伤活性和IFN-γ的产生(每组3只小鼠);皮下接种B16.F10黑色素瘤细胞(每组10只小鼠),观察荷瘤小鼠成活情况。结果AdhTRP2诱发小鼠CTL杀伤活性与免疫时间相关,免疫后1周活性达到最大,随后逐渐降低;AdhTRP2诱导的CTL杀伤活性具有剂量效应关系。AdhTRP2免疫小鼠1周其6hCTL杀伤率为94%±5%,脾脏产生IFNγ的CD8+T细胞占总CD8+T细胞的0.87%±0.12%,而相应地AdmTRP2分别为22%±8%和0.15%±0.05%。接种1×106B16.F10细胞,AdhTRP2免疫的小鼠观察3个月100%无瘤生长,但在病毒注射部位有白癜风形成;而接种1×105B16.F10细胞的AdmTRP2免疫小鼠3个月只有20%的成活率,并不伴局部白癜风形成。结论用Ad介导异种TRP2能有效地打破对自身TRP2的免疫耐受,诱发强烈的抗原特异性细胞免疫反应,同时伴发自身免疫性损伤。
Objective To investigate the relationship between anti-tumor immunity and autoimmune damage induced by adenovirus vector (Ad) mediated tyrosinase-related protein 2 (TRP2) -induced heterologous melanoma. Methods C57BL / 6 mice were immunized intradermally with Ad-encoding human or mouse TRP2 (AdhTRP2 or AdmTRP2). The mice were divided into normal control group (d1703), AdhTRP2 and AdmTRP2 groups. CTL killing activity and IFN-γ production (3 mice per group) were analyzed in vivo cytotoxic T lymphocyte (CTL) killing assay and intracellular interferon gamma (IFNγ) staining (ICS); B16.F10 melanoma Cells (10 mice in each group) to observe the survival of tumor-bearing mice. Results The cytotoxic activity of CTL induced by AdhTRP2 in mice was related to the immunization time. The activity of AdhTRP2 induced CTL was the highest at 1 week after immunization, and then decreased gradually. The CTL killing activity induced by AdhTRP2 was dose-dependent. AdhTRP2-immunized mice had a 6 hCTL killing rate of 94% ± 5% at 1 week and spleen IFNγ-producing CD8 + T cells at 0.87% ± 0.12% of total CD8 + T cells, whereas AdmTRP2 was 22% ± 8% 0.15% ± 0.05%. Inoculated with 1 × 106B16.F10 cells, mice immunized with AdhTRP2 showed 100% tumor-free growth for 3 months but had vitiligo formation at the site of virus injection. AdmTRP2-immunized mice vaccinated with 1 × 10 6 B 10 F 1 cells had only 20 % Of the survival rate, not with the formation of local vitiligo. Conclusion Adiponecting heterologous TRP2 with Ad can effectively break down the immune tolerance to TRP2 and induce a strong antigen-specific cellular immune response accompanied by autoimmune damage.