MUC1基因疫苗和GM-CSF对小鼠的细胞免疫和体液免疫应答  

Induction of specific T lymphocyte and humoral immune responses by MUC1 DNA vaccine and GM-CSF in mice

作　　者：袁时芳[1] 师长宏[2] 姚青[1] 李南林[1] 王廷[1] 王岭[1] 张英起[3] 

机构地区：[1]第四军医大学西京医院血管内分泌外科 [2]第四军医大学实验动物中心,陕西西安710033 [3]第四军医大学药学系生物技术中心,陕西西安710033

出　　处：《第四军医大学学报》2007年第17期1537-1540,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30571802);陕西省自然科学基金(2004C2-36)

摘　　要：目的:观察MUC1基因疫苗和GM-CSF共免疫对小鼠特异性细胞免疫和抗体水平的影响.方法:采用股四头肌肌肉注射,将构建的MUC1基因疫苗pcDNA3.1-MUC1免疫雌性Balb/c小鼠,每3wk1次,共3次.MUC1基因加GM-CSF组于每次基因免疫后1,3,5d,皮下注射GM-CSF100μL.最后1次基因免疫后第3周接种表达MUC1的EMT6小鼠乳腺癌细胞.2wk后观察、记录肿瘤的生长情况.流式细胞仪检测外周血CD4+和CD8+淋巴细胞亚群的百分比和比值.ELISA方法检测小鼠血清MUC1特异性抗体的水平.4h51Cr释放法检测小鼠脾特异性CTL的杀伤活性.结果:淋巴细胞亚群:与pcDNA3.1对照组相比,MUC1基因疫苗预防组的CD8+T淋巴细胞升高,差异有统计学意义(P<0.01),CD4+T淋巴细胞升高,差别无统计学意义;加GM-CSF佐剂预防组与单独MUC1疫苗预防组相比,CD4+T淋巴细胞升高,差异有统计学意义(P<0.01),CD8+T淋巴细胞升高差别无统计学意义.小鼠血清MUC1抗体水平:与对照组相比,MUC1基因疫苗预防组抗体水平升高,差异有统计学意义(P<0.05);加佐剂组与预防组相比,抗体水平升高但差别无统计学意义.在不同效靶比时,MUC1基因疫苗加GM-CSF组与单独MUC1基因免疫组相比较,特异性CTL对EMT6靶细胞的杀伤率差异显著(P<0.05).结论:MUC1基因疫苗可诱导小鼠产生特异性CD8+T淋巴细胞及体液免疫应答,GM-CSF对小鼠CD4+T淋巴细胞具有一定的调节作用.AIM： To observe the specific T lymphocyte responses and humoral immune responses against MUC1 in vivo induced by co-immunization with MUC1 DNA vaccine and GM-CSF in mice. METHODS： Female Balb/c mice were immunized intramuscnlarly with 100 mg MUC1 gene vaccine 3 times at 3-week intervals. On 1, 3 and 5 d after intra-muscular immunization, GM-CSF 100 μL （ 1μg/100μL） was given sc in MUC1 cDNA ＋ GM-CSF group. Three weeks after the last immunization, tumor challenge experiments were performed by using tumor cell line EMT6 expressing MUC1. Tumor growth inhibition was observed 2 weeks later. T-lymphocyte phenotype was detected by flow cytometry. MUCl-spocific antibody was detected by ELISA. CTL- specific cytotoxicity was detected by 4-hour 51Cr release assay. RESULTS： MUCl-specific CD8^＋ T cell responses were induced by MUC1 gene vaccine only （P 〈 0. 05 ）, compared to those of mice in pcDNA3. 1 control group; but CD4^＋ T cell responses were not significantly changed （ P 〉 0.05 ）. On the other hand, a dramatic augmentation of CD4^＋ T cell responses （ P 〈 0.05 ） was found with flow cytometry in the group of MUC1 cDNA ＋ GM- CSF （P 〈 0.01 ） ; however, CD8^＋ T cell responses were not significantly changed when compared to those of mice immunized with MUC1 gene vaccine only （ P 〉 0.05 ）. The levels of serum anti- MUC1 in the mice injected with MUC1 gene vaccine were significantly higher than those in pcDNA3.1 control group （P 〈 0.05）. The serum anti- MUC1 levels in the mice injected with the combination of MUC1 gene vaccine and GM-CSF were not significantly different from those with MUC1 gene vaccine only （ P 〉 0.05）. Compared with MUClcDNA group, the cytotoxicity of MUCl-spocific cytotoxic T lymphocytes to EMT6 target cells at various effector to target cell ratios was enhanced in the MUC1 DNA ＋ GM-CSF group （ P 〈 0.05）. CONCLUSION： MUC1specific CD8^＋ T cell responses and antibody production are elicited by MUC1 DNA immunization. And CD4^＋ T cel

关 键 词：MUC1粘蛋白 基因疫苗 粒细胞集落刺激因子 CD4^+ CD8^+ 体液免疫 

分 类 号：R392.11[医药卫生—免疫学]