热休克蛋白HSP65DNA疫苗抑制黑色素瘤血行转移的实验研究  

Study on HSP65 DNA Vaccine in the Hematogenous Metastasis of B16/F10 Melanoma

作　　者：林明[1] 鲁勇[1] 张宇[1] 谢燕飞[1] 胡向兵[1] 王华倩[1] 李泰明[1] 吴洁[1] 刘景晶[1] 曹荣月[1] 

机构地区：[1]中国药科大学生命科学与技术学院微基因药物实验室,江苏南京210009

出　　处：《药物生物技术》2009年第2期118-122,共5页Pharmaceutical Biotechnology

基　　金：国家青年自然科学基金(基金编号:30500458);国家自然科学基金(基金编号:30872393)资助

摘　　要：观察卡介苗来源的热休克蛋白HSP65的DNA疫苗对小鼠B16/F10黑色素瘤血行转移的抑制作用。构建含HSP65编码基因的真核分泌表达DNA疫苗pCR3.1-VS-HSP65及其后融合多个T细胞辅助表位的pCR3.1-VS-HSP65-TP-M2。连续8次肌肉注射免疫C57BL/6J雄性小鼠,采用ELISA法检测小鼠血清中抗HSP65-IgG类抗体滴度,于最后一次免疫后第2周,尾静脉接种B16/F10黑色素瘤细胞,考察该疫苗的免疫原性及药效。ELISA结果显示两种HSP65核酸疫苗均能诱发高滴度抗HSP-IgG类抗体。pCR3.1-VS-HSP65-TP-M2诱发抗体滴度较pCR3.1-VS-HSP65显著增加,并能显著抑制B16/F10的血行转移(P<0.05),且与生理盐水对照组对比抑瘤率达到74.8%(肝),53.2%(肺)。HSP65在增强其免疫原性的基础上能显著抑制B16/F10黑色素瘤的血行转移。To observe the inhibitory effect of the HSP65 DNA vaccine from Bacille Calmette Guerrin in the hematogenous metastasis of B16/F10 melanoma, two eukaryotic expression DNA vectors carrying the HSP65 encode gene were pCR3. 1-VS-HSP65 and pCR3. 1-VS-HSP65-TP-M2. Male C57BL/6J mice were continuously immunized intramuscularly 8 times in 8 weeks using above two DNA vaccines. The specific Anti-HSP65 antibody was detected by ELISA method. Two weeks after last immunization, B16/F10 melanomas cells were intravenously inoculated to investigate the immunogenicity and pharmacodynamic action of these vaccines. The results showed that the specific anti-HSP65 antibodies were detected in the antiserum of the male C57BL/6J mice immunized with the above two HSP65 DNA vaccines, pCR3. 1-VS-HSP65-TP-M2 could significantly induce the higher antibody titer than pCR3. 1-VS-HSP65 and significantly inhibit the hematogenous metastasis of B16/F10 melanoma. Tumor inhibitory rate was 74. 8% （liver） and 53. 2% （lung）compared with saline control group. HSP65 DNA vaccine can significantly suppress the hematogenous metastasis of B16/F10 melanoma on the basis of enhancing immunogenicity.

关 键 词：热休克蛋白 HSP65 DNA疫苗 黑色素瘤 抑瘤率 

分 类 号：R967[医药卫生—药理学]