人黑色素肿瘤相关抗原gp100质粒诱导特异性抗小鼠黑色素瘤的免疫保护效应

Human gp100 induces specific anti-murine melanoma immune response, implicating a potential way to antitumor immunotherapy

目的以人黑色素肿瘤相关抗原hgp100基因免疫小鼠,诱导针对小鼠黑色素瘤的特异性免疫应答及保护效应。方法分别构建含全长人gp100(hgp100)和小鼠印100(mgp100)编码基因的质粒pcDNA3-hgp100和pcDNA3-mgp100,以100μg DNA肌肉免疫C57BL/6小鼠3次,每次间隔2周。通过体外免疫功能及体内攻击实验评价hgp100诱导对小鼠黑色素瘤的特异性免疫应答及保护效应。结果与mgp100比较,hgp100能诱导特异性针对mgp100的抗体生成和T细胞增殖及杀伤,产生以 IFN-γ分泌为主的特异性免疫应答;hgp100免疫小鼠在小鼠黑色素肿瘤细胞攻击后生存时间显著延长。结论人黑色素瘤相关抗原gp100可诱导特异性抗小鼠黑色素瘤的免疫应答及保护效应。

Objective To investigate the efficacy of hgp100, a human melanoma associate antigen, in inducing specific anfi-murine melanoma immune response and in protection via hgp100 plasmid. Methods Plasmids pcDNA3-hgp100 and pcDNA3-mgp100 were constructed by using full-length hgp100 and mgp100 coding sequence inserted into pcDNA3 vectors respectively. C57BL/6 mice were immunized intramuscularly with pcDNA3, pcDNA3-mgp100 and pcDNA3-hgp100 three times at two week intervals, separately. Specific anti-tumor antibody derived from immunized serum was detected kinetically by ELISA. Specific proliferation and cytotoxicity against B16F10 were detected. IFN-γ and IL-4 cytokines were measured. Mice immunized with plasmida were finally challenged with B16F10 melanoma tumor to observe the functional protection. Results Mice immunized with pcDNA3-hgp100 exhibited a significant protection （survival as long as 68 days） as compared to the mice immunized with pcDNA3-mgp100 only （survival to 29 days）. Both T cell proliferation and cytotoxicity against murine melanoma were enhanced by hgp100 DNA immunization. An increased IFN-γ release and antibody production could also be detected after immunization with the pcDNA3-hgp100. Conclusion The data strongly indicated that anti-tumor immune response and protection against murine melanoma could be induced by human gp100, implicating a potential approach in anti-tumor immunotherapy.