BCR-ABL-SEA DNA疫苗诱导BALB/c小鼠的免疫应答  

Immune responses induced by DNA vaccine of BCR-ABL-SEA in BALB/c mice

作　　者：高永鹏[1] 林晨[1] 田红霞[1] 陈琛[1] 秦雅楠[1] 周羽竝[2] 李扬秋[3] 

机构地区：[1]暨南大学医学院微生物与免疫学系,广东广州510632 [2]暨南大学医学院生物化学系,广东广州510632 [3]暨南大学医学院血液病研究所,广东广州510632

出　　处：《中国病理生理杂志》2011年第2期361-366,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.06025169);广州市科技支撑计划资助项目(No.2009Z1-E161)

摘　　要：目的:了解BCR-ABL-SEA双表达DNA疫苗诱导BALB/c小鼠特异性细胞和体液免疫应答效应。方法:用已成功构建的重组双表达BCR-ABL多肽和SEA多肽的质粒BCR-ABL-pIRES-SEA(B-P-S)免疫小鼠,间隔14 d共3次。相同方法用单表达BCR-ABL多肽或SEA多肽的质粒BCR-ABL-pIRES和SEA-pIRES免疫小鼠作对照。利用CCK-8比色法检测小鼠脾脏T细胞对K562细胞株的杀伤活性;流式细胞术测定小鼠脾脏CD4+与CD8+T细胞表达情况;ELISA法检测小鼠血清中干扰素γ(IFN-γ)和白细胞介素4(IL-4)生成情况;间接免疫荧光法检测血清中抗BCR-ABL抗体。结果:免疫后第7周时,双表达重组质粒B-P-S组小鼠脾脏CTL细胞针对K562杀伤率、血清中INF-γ含量均明显高于单表达BCR-ABL-pIRES组和SEA-pIRES组(P<0.05);CD4+/CD8+T细胞比值、血清中IL-4含量各组之间无明显差异(P>0.05);荧光显微镜检测到血清中有抗BCR-ABL抗体。结论:所构建的BCR-ABL-SEA重组双表达质粒可诱导小鼠产生特异性细胞和体液免疫应答效应。AIM： To evaluate the immune responses induced by a DNA vaccine expressing both BCR - ABL and SEA peptides in mice. METHODS： The vaccine plasmid of BCR - ABL - plRES - SEA, which expressed both BCR - ABL and SEA peptides and was constructed previously in our laboratory, was intramuscularly injected into BALB/c mice at 14 - day intervals for 3 cycles. The plasmids of BCR - ABL - pIRES and SEA - plRES, which were only expressed ei- ther BCR/ABL peptide or SEA peptide, were also used in the same procedure for comparison. The cytotoxicity of T - cells from mouse spleen against K562 cell line was examined by cell counting kit - 8 （ CCK - 8）. The ratio of CD4＋ to CD8＋ on the harvested T cells was detected by flow cytometry. The serum levels of interferon -γ（ IFN- γ） and interleukin 4（ IL- were measured by ELISA. The antibodies against BCR - ABL were detected by the technique of RESULTS ： Seven weeks after immunization, the specific cytotoxicity of T - cells against K562 cells and the level of INF - γ in co - expression of BCR - ABL - pIRES - SEA group were significantly higher than those in mono - expression of BCR -ABL- plRES group and SEA- pIRES group（ P 〈0. 05 ）. The ratio of CIM ＋ to CD8 ＋ on the harvested T- cells and the level of IL-4 in vaccinated mouse serum were not significantly different among the groups（P 〉 0. 05 ）. The specific anti- body against BCR - ABL was detected in BCR - ABL - pIRES - SEA group and BCR - ABL - plRES group but not in SEA - pIRES group by indirect immunofluorescene analysis. CONCLUSION： The recombinant BCR - ABL - plRES - SEA vaccine induces specific humoral and cellular immune responses in vivo.

关 键 词：白血病 基因 BCR—ABL融合 葡萄球菌肠毒素A 疫苗 DNA 

分 类 号：R363[医药卫生—病理学]