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目的观察重组弓形虫磷酸甘油酸变位酶2(rTgPGAM2)联合IL-2或IFN-γ佐剂滴鼻免疫小鼠诱导的特异性抗体免疫应答。方法将48只6周龄BALB/c小鼠随机均分为6组:免疫组分别用500IU IL-2、1 000UIFN-γ、30μg rTgPGAM2、30μg rTgPGAM2+500IU IL-2、30μg rTgPGAM2+1 000UIFN-γ滴鼻免疫,抗原和佐剂均溶于20μl PBS中,对照组滴鼻20μl PBS。共免疫3次,第1次免疫与第2次免疫间隔14d,第2次免疫与第3次免疫间隔7d。末次免疫后第14d,颈椎脱臼处死小鼠,ELISA法检测血清IgA和IgG及鼻咽冲洗液、阴道冲洗液、肠冲洗液sIgA水平。结果免疫组血清IgA和IgG水平高于对照组,以rTgPGAM2+IL-2组和rTgPGAM2+IFN-γ组升高更显著(F=18.653和8.853,P<0.05);免疫组小鼠鼻咽冲洗液、阴道冲洗液sIgA水平显著高于对照组(F=6.670和7.288,P<0.05),rTgPGAM2+IL-2和rTgPGAM 2+IFN-γ免疫组小鼠小肠冲洗液sIgA水平显著高于对照组(F=4.557,P<0.05)。结论单独用rTgPGAM2或佐剂或两者联合免疫小鼠均能诱导黏膜特异性体液免疫应答,其中rTg-PGAM2联合IL-2佐剂能诱导产生更高水平的抗体。
Objective To observe the specific immune response induced by recombinant human Toxoplasma gondii phosphoglycerate mutase 2 (rTgPGAM2) combined with IL-2 or IFN-γ adjuvant intranasal immunization. Methods 48 6-week-old BALB / c mice were randomly divided into 6 groups: immunized with 500IU IL-2,1 000UIFN-γ, 30μg rTgPGAM2, 30μg rTgPGAM2 + 500IU IL-2, 30μg rTgPGAM2 + 1 000UIFN- γ intranasal immunization, antigen and adjuvant were dissolved in 20μl PBS, the control group intranasal 20μl PBS. Immunization with three times, the first immunization and the second immunization interval 14d, the second immunization and the third immunization interval 7d. At the 14th day after the last immunization, the mice were sacrificed by cervical dislocation, and serum IgA and IgG, nasal pharyngeal fluid, vaginal fluid and intestinal fluid sIgA were detected by ELISA. Results The levels of serum IgA and IgG in the immunized group were significantly higher than those in the control group, and were significantly higher in the rTgPGAM2 + IL-2 and rTgPGAM2 + IFN-γ groups (F = 18.653 and 8.853, P <0.05) The sIgA level of liquid and vaginal washes was significantly higher than that of the control group (F = 6.670 and 7.288, P <0.05). The levels of sIgA in rTgPGAM2 + IL-2 and rTgPGAM 2 + IFN- (F = 4.557, P <0.05). CONCLUSION: Mucosal-specific humoral immune responses can be induced by rTgPGAM2 or adjuvant alone or in combination with both. The rTg-PGAM2 combined with IL-2 adjuvant can induce higher levels of antibodies.