弓形虫表面抗原P30 DNA疫苗免疫小鼠诱导细胞免疫应答的研究

STUDY ON THE CELLULAR IMMUNE RESPONSES INDUCED BY P30 DNA VACCINE OF TOXOPLASMA GONDII IN MICE

目的 观察弓形虫表面抗原 P30 DNA疫苗诱导小鼠产生的细胞免疫应答。 方法 大量制备重组真核表达质粒 p BK- P30 ,用生理盐水 (NS)稀释至 1μg/μl,1次肌注免疫 BAL B/ c小鼠 ,分别在免疫后 5和 10周 ,通过 Con A刺激 ,MTT法测定免疫鼠脾脏 T淋巴细胞转化率 ;使用间接免疫荧光法 ,用流式细胞仪对 CD4+、CD8+T细胞亚群进行测定。 结果Con A刺激小鼠脾 T淋巴细胞发生增殖反应 ,p BK- P30免疫组与两对照组及对照组之间的差异均无显著性 (P>0 .0 5 )。T细胞亚群 CD4+、CD8+动态分析 ,CD4+T细胞在各组增殖均不明显 (P>0 .0 5 ) ,但免疫组 CD8+细胞数量升高 ,CD4+/CD8+比率下降 ,与空质粒 p BK- CMV对照组和 NS对照组之间差异均有显著性 (P<0 .0 5 ,P<0 .0 1) ,p BK- CMV对照组与 NS对照组之间差异也有显著性 (P<0 .0 1)。免疫后 10周 ,与 NS对照组相比较 ,免疫组和 p BK- CMV对照组 CD8+细胞仍升高 (P<0 .0 1) ,但它们之间的差异无显著性 (P>0 .0 5 )。 结论 弓形虫表面抗原 P30 DNA疫苗能诱导 BAL B/

Objective To observe the cellular immune response induced by P30 DNA vaccine of Toxoplasma gondii in mice. Methods Large scale preparation of pBK P30 was dissolved to 1 μg/μl using normal saline and injected into BALB/c mice via muscles, the mice were immunized onetime. After 5 and 10 weeks immunization, the proliferation rates of spleen cells, stimulated by ConA, were detected by means of MTT assay respectively. With the help of flow cytometer(FCM), T cell subgroup CD 4 + and CD 8 + were tested by the indirect immune fluorescent assay. Results The proliferating response of all group of mouse spleen T cells were generated when stimulated by ConA, but there was no remarkable difference among control mice and immunized ones ( P >0.05). The kinetic analysis of CD 4 + and CD 8 + T cell subgroup showed that the number of CD 4 + T cells had no obvious increase in all groups, but CD 8 + T cells in immunized mice were obviously increased and the rates of CD 4 +/CD 8 + were decreased compared with the pBK CMV control and NS control ones after 5 weeks immunization respectively ( P <0.05, P <0.01). There was remarkable difference between pBK CMV and NS control group ( P <0.01). After 10 weeks immunization, CD 8 + T cells of immunized group and pBK CMV control were still increased compared with NS control group( P <0.01), but there was no remarkable difference between them( P >0.05). Conclusion The P30 DNA vaccine of T. gondii could induce the cellular immune response in BALB/c mice.