【摘要】 目的:采用黏蛋白与麦芽糖结合蛋白融合蛋白(MUC1-MBP)作为特异性抗原,研究胸腺肽α1(Tα1)加强诱导MUC1特异性免疫应答的类型,探讨其作为佐剂应用的可行性。方法:将C57BL/6小鼠随机分为生理盐水组(注射生理盐水)、MUC1-MBP+BCG组(注射MUC1-MBP+BCG)和MUC1-MBP和Tα1组(注射MUC1-MBP+Tα1),分别进行T细胞免疫活性、MUC1特异性抗体效价及亚类、Tα1联合MUC1-MBP抗肿瘤作用检测。第3次免疫后4~7d无菌取脾脏组织,测定脾指数;淋巴细胞增殖反应测定各组小鼠的刺激指数(SI);ELISA法测定小鼠脾细胞培养上清中干扰素γ(IFN-γ)、白细胞介素2(IL-2)、白细胞介素4(IL-4)和白细胞介素10(IL-10)的水平及小鼠血清中MUC1特异性抗体水平;第3次免疫后7d,注射黑色素瘤细胞B16-MUC1,观察各组小鼠生存期。结果:与生理盐水组比较,MUC1-MBP+BCG组和MUC1-MBP+Tα1组小鼠脾指数与SI均升高(P<0.05或P<0.01),MUC1特异性SI明显升高(P<0.05)。与生理盐水组比较,MUC1-MBP+BCG组小鼠脾细胞培养上清中IFN-γ和IL-2水平均明显升高(P<0.05);IL-4和IL-10水平均略微升高,但差异无统计学意义(P>0.05)。与生理盐水组比较,MUC1-MBP+Tα1组小鼠脾细胞培养上清中IL-4水平明显升高(P<0.01);IFN-γ、IL-2和IL-10水平均略微升高,但差异无统计学意义(P>0.05)。MUC1-MBP+Tα1免疫小鼠能够产生抗MUC1特异性抗体且效价随浓度升高而升高。抗体亚型检测,与生理盐水组比较,MUC1-MBP+Tα1组IgG1水平明显升高(P<0.05或P<0.01),IgG2a水平无明显变化。肿瘤预防实验,与生理盐水组比较,MUC1-MBP+BCG组和MUC1-MBP+Tα1组小鼠生存期未见明显差异。结论:MUC1-MBP联合Tα1更倾向于Th2型免疫应答,Tα1可以作为预防性疫苗佐剂使用,不适合治疗性疫苗使用。更多还原

【Abstract】 Objective:To investigate the MUC1 specific immune response enhanced by thmosinα1(Tα1)using MUC1-MBP as the specific antigen,and to discuss the feasibility of MUC1-MBP as an adjuvant.Methods:The C57 BL/6 mice were randomly divided into normal saline group,MUC1-MBP+BCG group and MUC1-MBP+Tα1 group.The T cellular immune activities,MUC1 special antibody and subclass,anti-tumor effect of Tα1 combined with MUC1-MBP were detected.4-7 dafter the 3 rd immunization,the spleen indexes of the mice in various groups were measured;the lymphocyte proliferation response was used to detect the stimulate index(SI)of the mice in various groups;the levels of specific cytokines IFN-γ,IL-2,IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA;the level of serum MUC1-specific antibody was detected by ELISA.The C57 BL/6 mice were inoculated with B16-MUC1 7 dafter the last immunization and the survival of the mice was observed.Results:Compared with normal saline group,the spleen index and SI of the mice in MUC1-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased(P< 0.05 or P<0.01);the specific SI of MUC1 were significantly increased(P< 0.05).Compared with normal saline group,the levels of IFN-γand IL-2 in the supernatant of spleen cells of the mice in MUC1-MBP+BCG group were obviously increased(P<0.05);the levels of IL-4 and IL-10 were slightly increased,but there were no significant differences(P>0.05);compared with normal saline group,the level of IL-4 in MUC1-MBP+Tα1 group was obviously increased(P< 0.01);the levels of IFN-γ,IL-2 and IL-10 were slightly increased,but there were no significant differences(P>0.05).The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1.The antibody subtype detection results showed that compared with normal saline group,the level of IgG1 in MUC1-MBP+BCG group was significantly increased(P < 0.05 or P < 0.01),and the level of IgG2 ahad no obvious change.The tumor prevention experiment results showed that compared with normal saline group,the survival rates of the mice in MUC1-MBP+BCG group and MUC1-MBP+Tα1 group had no significant differences.Conclusion:MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice.It indicates that Tα1 can be used as an adjuvanted preventive vaccines,but not suitable for therapeutic vaccines.更多还原