人黏蛋白1串联重复区DNA疫苗诱导小鼠的免疫应答

Immune Response Induced by MUC1 VNTR DNA Vaccine in Mice

目的观察人黏蛋白1串联重复区(MUC1 VNTR)DNA疫苗诱导BALB/c小鼠的体液及细胞免疫应答。方法将含有33个黏蛋白1重复区序列(33m)的重组质粒pVR1012-33m,经肌肉注射免疫BALB/c小鼠,并加强免疫2次,每次100μg,间隔2周,分别以空载体pVR1012和生理盐水为对照。末次免疫后取小鼠血清及脾淋巴细胞,Western blot检测血清抗体特异性,LDH法检测CTL反应活性,MTS/PMS法检测脾淋巴细胞增殖活性。结果重组质粒免疫组小鼠血清中检出了MUC1 VNTR抗原特异性抗体。用重组质粒pVR1012-33m免疫BALB/c小鼠后,在效靶比为100:1和33:1时,可显著地诱导特异性CTL应答。在MUC1 VNTR抗原肽的刺激下,免疫小鼠脾淋巴细胞得到增殖,与空载体和生理盐水对照组相比,差异均有显著意义。结论DNA疫苗pVR1012-33m在BALB/c小鼠体内可诱导特异性细胞免疫和体液免疫应答,为预防、治疗性MUC1 VNTR疫苗的研制提供了一定的实验依据。

Objective To observe the humoral and cellular immune responses induced by MUC1 variable number of tandem repeats,VNTR DNA vaccine in mice. Methods Inject i. m. BALB/c mice with 100 p.g of recombinant plasmid pVR1012-33m contai- ning 33 repeats of MUC1 VNTR for 3 times, at an interval of 2 weeks, using empty vector pVR1012 and physiological saline as control. Collect the sera and spleen lymphocytes of mice after immunization and test for serum antibody specificity by Western blot ,CTL activity by LDH method,and spleen lymphocyte proliferation activity by MTS/PMS method. Results Specific antibody against MUC1 VNTR antigen was detected in sera of mice immunized with recombinant plasmid pVR1012-33m. When the ratio of effector cells to target cells were 100：1 and 33： 1, specific CTL responses were induced in immunized mice significantly. Compared with those immunized with emp- ty vector and physiological saline ,the spleen lymphocythes of mice immunized with recombinant plasmid pVR1012-33m proliferated sig- nificantly. Conclusion Recombinant plasmid pVR1012-33m induced specific cellular and humoral immune responses in mice, which provided experimental basis for the development of prophylactic and therapeutic MUCI VNTR vaccine.