牛环形泰勒虫烯醇化酶重组蛋白对小鼠免疫应答的研究OACSTPCD

The humoral immunity level and cellular immunity level of experimental animals were evaluated by using the prokaryotic recombinant protein of Theileria annulata enolase(2-phospho-D-glyceratehydrolyase,enolase).Then the reactivity of rTa-enolase was also estimated by using Western blot after rTa-enolase was purified and concentrated using"His"purification column and ultrafiltration tube.In addition,polyclonal antibodies of rTa-eno-lase were prepared due to immunized Balb/c mice,and the specific antibody level of rTa-enolase was measured by indirect ELISA.The splenic T-cell subpopulations were also tested by flow cytometry which were used for prepared lymphocyte suspensions of immunized mouse spleens.Furthermore,proliferation of lymphocytes was examined by MTT kit after in vitro stimulation.The data were statistically analyzed by one-way ANOVA.Western blot analysis proved rTa-enolase can specifically react with positive sera,while the significant differences(P＜0.01)were ob-served in ELISA.The ratio of CD4+/CD8+T cells in the immunized group(2.53±0.11)were significant differ-ences(P＜0.01)compared with control group(1.85±0.11).Proliferation profile of spleen lymphocytes were signif-icant induced after mouse spleen lymphocytes was stimulated by rTa-enolase(P＜0.05).The results of this study indicated that high level of humoral immunity and cellular immune response was identified after rTa-enolase im-munization of mice,especially CD4+T cell.This study will be helpful for its subsequent immune function in vac-cines.