【摘要】 目的探讨阴道毛滴虫重组AP33融合蛋白对宿主的保护性免疫机制。方法用纯化的重组AP33蛋白抗原免疫BALB/c小鼠,检测小鼠血清和脾细胞多项免疫指标的变化;实验组与对照组分别用滴虫滋养体进行皮下和腹腔攻击,观察各组小鼠的行为变化和死亡时间。结果重组AP33蛋白免疫小鼠产生高效价抗体;实验组小鼠的CD4+T淋巴细胞有所增高(30.65±3.69)%,CD4+/CD8+比值明显升高(3.96±0.56)%(P<0.01);脾细胞培养上清液中mIL-2、mIFN-γ、mIL-4和mIL-6均有不同程度的升高,尤其mIFN-γ浓度增高最明显;皮下注射组,实验组小鼠背部出现明显溃疡;腹腔注射组,对照组小鼠6d内全部死亡,而实验组小鼠的存活时间比对照组长,其差异有统计学意义(P<0.05)。结论重组AP33融合蛋白具有较强的免疫原性,能诱导小鼠产生较好的免疫保护作用,可能成为阴道毛滴虫预防或治疗性多价疫苗的侯选抗原。更多还原

【Abstract】 To investigate the immuno-protective mechanisms of the recombinant protein AP33 from Trichomonas vaginalis,the purified recombinant protein AP33 from Trichmonas vaginalis was used as antigen to immunize 36 BALB/c mice subcutaneously for 3 times; 1-2weeks after the final immunization,IgA and IgG antibodies in sera were measured by ELISA,and the specific CD+4 and CD+8 T cells were analyzed by flow cytometry(FCM). The supernatants of cultured spleen cells of 10 mice in each group were tested by sandwich ELISA assay to determine the presence of mouse IFN-γ,IL-2,IL-4 and IL-6. Eight mice in each group were challenged subcutaneously and intraperitoneally with 2 × 106 T.vaginalis,and their change in behaviors and survival times were thereby observed. It was found that mice immunized with recombinant AP33 protein produced high-titered antibodies with increased number of the CD+4 T lymphocytes (30.65 ± 3.69%) and elevated ratio of CD+4/CD+8 T cells (3.96 ± 0,56%) (P<0.01). The contents of cytokines IL-2,IFN-γ. IL-4 and IL-6 in the supernatants of mouse spleen cell cultures elevated in different degrees,especially the level of IFN-γ in the supernatants of cultured spleen cells of the immunized mice (453.15 ± 60.78 μg/ml)(P<0.01). After subcutaneous chanllenge with T.vaginalis ,many skin ulcers occurred on the back of mice,while after intraperitoneal challenge,the survival time in the experimental group was longer than that of the control group of mice with statistical difference (P<0.05). It is concluded that the recombinant fusion protein AP33 has strong immunogenicity which can induce better immuno-protection response against infection with T.vaginalis ,and should serve as a candidate antigen for the preparation of the therapeutic poly-vaccine.更多还原