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目的观察重组弓形虫磷酸甘油酸变位酶2(Recombinant Toxoplasma gondii phosphoglycerate mutase 2,rTgPGAM2)联合白细胞介素-2(Interleukin-2,IL-2)或干扰素γ(Interferonγ,IFNγ)滴鼻免疫小鼠诱导的脾淋巴细胞免疫应答,探讨IL-2和IFNγ的佐剂效应。方法将48只BALB/c小鼠随机均分为6组:rTgPGAM2组(30μg)、IL-2组(500 IU)、IFNγ组(1 000 IU)、rTgPGAM2(30μg)+IL-2(500 IU)组和rTgPGAM2(30μg)+IFNγ(1 000 IU)组和对照组(20μl PBS),免疫途径均为滴鼻免疫,共免疫3次。末次免疫后第14天,颈椎脱臼处死小鼠,CCK-8法检测各组小鼠脾淋巴细胞增殖活性,ELISA法检测脾淋巴细胞培养上清中IL-2、IFNγ、IL-4和IL-10水平。结果经rTgPGAM2刺激后,与对照组比较,各免疫组小鼠脾淋巴细胞刺激指数(Stimulation index,SI)均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);经ConA刺激后,仅IL-2组SI显著高于对照组(P<0.01)。各免疫组小鼠脾淋巴细胞培养上清中IL-2和IFNγ含量均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);rTgPGAM2组及其联合IL-2或IFNγ组IL-4含量显著高于对照组(P<0.05);而IL-10的含量,只有IFNγ组和rTgPGAM2+IFNγ组显著高于对照组(P<0.05)。结论 rTgPGAM2联合IL-2或IFNγ鼻内免疫小鼠诱导的脾淋巴细胞免疫应答优于rTgPGAM2单独免疫,表明IL-2和IFNγ具有良好的佐剂效应,IL-2的佐剂效应更佳。
Objective To observe the effect of recombinant Toxoplasma gondii phosphoglycerate mutase 2 (rTgPGAM2) combined with Interleukin-2 (IL-2) or Interferon γ (IFNγ) Induced immune response of splenic lymphocytes, to explore the adjuvant effect of IL-2 and IFNγ. Methods 48 BALB / c mice were randomly divided into 6 groups: rTgPGAM2 group (30μg), IL-2 group (500IU), IFNγ group (1 000IU), rTgPGAM2 (30μg) + IL-2 ) Group and rTgPGAM2 (30μg) + IFNγ (1 000 IU) group and control group (20μl PBS) were immunized intranasal immunized three times. On the 14th day after the last immunization, mice were sacrificed by cervical dislocation. The proliferation activity of splenic lymphocytes was detected by CCK-8 assay. The levels of IL-2, IFNγ, IL-4 and IL- 10 levels. Results After stimulation with rTgPGAM2, compared with the control group, the spleen lymphocyte stimulating index (SI) in each immunized group was significantly higher than that in the control group (P <0.05 or P <0.01), and rTgPGAM2 combined with IL-2 or IFNγ Group was significantly higher than that of rTgPGAM2 group (P <0.01 or P <0.05). After stimulation with ConA, SI in IL-2 group was significantly higher than that in control group (P <0.01). The levels of IL-2 and IFNγ in the splenic lymphocyte culture supernatant of each immunized group were significantly higher than that of the control group (P <0.05 or P <0.01), and rTgPGAM2 combined with IL-2 or IFNγ group was significantly higher than that of the rTgPGAM2 group (P < 0.01 or P <0.05). The content of IL-4 in rTgPGAM2 group and its combination with IL-2 or IFNγ group was significantly higher than that in control group (P <0.05), while the content of IL-10 in IFNγ group and rTgPGAM2 + In the control group (P <0.05). Conclusions The immune response of spleen lymphocytes induced by rTgPGAM2 combined with intranasal immunization with IL-2 or IFNγ is better than that of rTgPGAM2 alone, indicating that IL-2 and IFNγ have good adjuvant effect and IL-2 adjuvant effect is better.