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目的动态观察弓形虫可溶性速殖子抗原(soluble tachyzoite antigen,STAg)滴鼻和皮下免疫小鼠诱导的黏膜免疫和系统免疫应答。方法6周龄BALB/c小鼠90只随机分为3组,分别以20μg STAg滴鼻或皮下注射免疫小鼠2次,间隔2周,对照组不做处理。末次免疫后1、2、3、4和5周,每组随机处死6只小鼠。ELISA法测定小肠冲洗液sI-gA和血清IgG,分离并计数小肠上皮内淋巴细胞(intraepithelial lymphocyte,IEL)和脾淋巴细胞。结果对照组小鼠小肠冲洗液sIgA水平和IEL数及血清IgG水平和脾淋巴细胞数均维持在较低水平。滴鼻免疫组小鼠小肠冲洗液sIgA水平升高,与对照组比较差异有统计学意义(FsIgA=10.074,FIEL=14.747,P<0.01),其中第5周sIgA水平、第2~5周IEL数量与皮下免疫组比较差异有统计学意义(FsIgA=7.862,FIEL=9.807,P<0.05)。皮下免疫组小鼠血清IgG水平和脾淋巴细胞数均升高,与对照组比较差异有统计学意义(FIgG=8.207,F脾细胞=11.209,P<0.05),第5周脾淋巴细胞数与滴鼻组比较差异有统计学意义(F=6.826,P<0.05)。结论20μg STAg滴鼻免疫小鼠诱导的黏膜免疫应答水平优于同剂量的皮下免疫,同时也诱导了较高水平的系统免疫应答。皮下免疫可诱导较高水平的系统免疫应答,但其诱导的黏膜免疫应答较弱。
Objective To observe the mucosal and systemic immune responses induced by intranasal and subcutaneous immunization of soluble Tachyzoite antigen (STAg) in mice. Methods 90 BALB / c mice of 6 weeks old were randomly divided into 3 groups. The mice were immunized with 20μg STAg intranasally or subcutaneously twice a week for 2 weeks without any treatment in the control group. One, two, three, four and five weeks after the last immunization, 6 mice were randomly sacrificed in each group. Small intestine washing solution sI-gA and serum IgG were measured by ELISA. Small intestine intraepithelial lymphocyte (IEL) and spleen lymphocytes were isolated and counted. Results The sIgA level, IEL number, serum IgG level and splenic lymphocyte count in the control group mice were all kept at a low level. The sIgA level in intestinal mucosa of mice immunized with intranasal immunization group was significantly higher than that in control group (FsIgA = 10.074, FIEL = 14.747, P <0.01). The levels of sIgA in week 5, IEL There was significant difference between the quantity and subcutaneous immunization group (FsIgA = 7.862, FIEL = 9.807, P <0.05). The level of serum IgG and the number of splenic lymphocytes in subcutaneous immunized mice were significantly higher than those in control group (FIgG = 8.207, F splenocytes = 11.209, P <0.05) Compared with the nasal drops group, the difference was statistically significant (F = 6.826, P <0.05). Conclusion The mucosal immune response induced by 20μg STAg intranasal immunization mice is superior to the same dose of subcutaneous immunization, but also induced a higher level of systemic immune response. Subcutaneous immunization can induce a higher level of systemic immune response, but its mucosal immune response is weaker.