腺病毒编码人TRP2修饰DC诱发抗小鼠黑色素瘤免疫的研究  

Immunity against murine melanomas elicited by dendritic cells infected with adenovirus encoding human tyrosine-related protein 2

作　　者：谭晓华 [1] Wan Yonghong  

机构地区：[1]100700,北京军区总医院血液科 [2]Centre for Genqe Therapy,Molecular and Pathological Medicine Department,Medical Centre,McMaster University,Hamilton,ON.Canada,L8S

出　　处：《中华微生物学和免疫学杂志》2005年第12期993-997,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目(30271185和30371627)

摘　　要：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 difference in immunity against routine melanomas elicited by dendritic cells infected with adenovirus encoding human or routine tyrosinase-related protein 2 （Ad hTRP2 or Ad m TRP2）. Methods Analysis of the killing activity and production of IFN-γ by antigen-specific cytotoxic T lymphocytes（CTL） from the C57BL/6 mice immunized subcutaneously with bone morrow-derived dendritic cells（ BM- DC） infected with adenovirus encoding human or murine TRP2 （Ad hTRP2/BM-DC or Ad mTRP2 BM-DC） were made using in vivo CTL and intracellular staining of IFN-γ（ICS）, respectively. Additionally, the survival rate of immunized mice was checked after the subcutaneous inoculation with murine melanoma B 16 . F 1 0 cells . Results It was shown by in vivo CTL and ICS that, in the spleen, 6 h CTL killing and IFN-γ-predueing CD8^＋ T cells in total CD8^＋ T cells induced by Ad hTRP2/BM-DC were （98.7 ± 1.2）% and （1.25 ± 0.21）%, respectively, whereas Ad mTRP2/BM-DC-induced 6 h CrL killing and IFN-γ-producing CD8^＋ T cells were just （28.6± 6.3）% and （0.24±0.06）%, respectively. Moreover, 100% of the Ad hTRP2/BM-DC-immunized mice inoculated subcutaneously with 10^6 B16.F10 cells were in a tumor-free state for three months, whereas only 40% of the Ad mTRP2/BM-DC-immunized mice with 5 × 10^4 B16. F10 cells were survived. Conclusion BM-DC modified with Ad hTRP2 breaks immune tolerance more effectively to murine melanomas and induces stronger immunity against melanomas as compared to BM-DC modified with Ad mTRP2, therefore acting as a highly effective dendritic cell-based tumor vaccine.

关 键 词：人腺病毒 树突状细胞 一元酚单氨酶 嗜异性抗原 免疫耐受 

分 类 号：R73-3[医药卫生—肿瘤]