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目的 :观察MUC1基因疫苗诱导小鼠特异性杀伤性T细胞及体液免疫应答的作用。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 ,每次间隔 3wk ,共 3次。最后 1次免疫后第 3周 ,接种表达MUC1的EMT6乳腺癌细胞进行免疫保护实验。用 4h51Cr释放法检测小鼠脾细胞特异性CTL杀伤活性 ;免疫组化染色法检测小鼠血清特异性抗体的水平。结果 :在效靶比为 10 0∶1、5 0∶1、2 5∶1、12 .5∶1时 ,MUC1基因疫苗免疫组特异性CTL对EMT6靶细胞杀伤活性分别为 5 4 .1%、39.8%、2 6 .4 %和2 0 .1% ,对照组分别为 13.2 %、10 .0 %、8.2 %、7.2 %和 11.7%、9.8%、7.7%、7.0 % ,前者与后二者差异显著 (P <0 .0 1)。免疫组化染色检测显示 ,人乳腺癌组织MUC1呈染色阳性 ;MUC1基因疫苗免疫组仅见 4 0 % (4/ 10 )的小鼠有肿瘤形成 ,而 pcDNA3.1对照组和生理盐水阴性对照组 10 0 %可见肿瘤形成、生长 ,表明MUC1基因疫苗免疫组小鼠具有一定的免疫保护作用。结论 :MUC1基因疫苗可诱导小鼠产生特异性CTL及体液免疫应答 ,对小鼠体内荷瘤可能具有一定的预防作用
Objective: To observe the effect of MUC1 gene vaccine on specific killer T cells and humoral immune response in mice. Methods: Female Mice were immunized with the MUC1 gene vaccine pcDNA3.1 MUC1 by intramuscular injection of the femoral quadriceps, three times at 3wk intervals. At the third week after the last immunization, MUC1-expressing EMT6 breast cancer cells were inoculated for immunopreservation experiments. 4h51Cr release method was used to detect the specific CTL cytotoxic activity of mouse spleen cells; immunohistochemistry was used to detect the level of serum specific antibody in mice. Results: When the target ratio was 100: 1, 50: 1, 25: 1, 12: 5: 1, the killing activity of MUC1 gene specific CTL on EMT6 target cells was 54.1% , 39.8%, 26.4% and 20.1% in the control group, and 13.2%, 10.0%, 8.2%, 7.2% and 11.7%, 9.8%, 7.7% and 7.0% The difference was significant (P <0.01). Immunohistochemical staining showed that MUC1 was stained positive in human breast cancer tissues. Only 40% (4/10) mice in the MUC1 vaccine group showed tumor formation, whereas the pcDNA3.1 control group and saline-negative control group 10 0% showed tumor formation and growth, indicating MUC1 gene vaccine immunized mice have a certain degree of immune protection. Conclusion: MUC1 gene vaccine can induce specific CTL and humoral immune responses in mice, which may have some preventive effects on tumor-bearing in mice