【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。更多还原

【Abstract】 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-pIRES-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-pIRES,which were only expressed either 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-4) were measured by ELISA.The antibodies against BCR-ABL were detected by the technique of immunofluorescence. 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-pIRES group and SEA-pIRES group(P<0.05).The ratio of CD4+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 antibody against BCR-ABL was detected in BCR-ABL-pIRES-SEA group and BCR-ABL-pIRES group but not in SEA-pIRES group by indirect immunofluorescene analysis. CONCLUSION: The recombinant BCR-ABL-pIRES-SEA vaccine induces specific humoral and cellular immune responses in vivo.更多还原