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目的探讨可溶性速殖子抗原(STAg)联合蜂胶佐剂鼻内免疫小鼠抗弓形虫感染的保护作用及蜂胶的适宜剂量。方法将60只BALB/c小鼠随机分为5组,每组12只,分别用20μgSTAg、20μgSTAg+5μg蜂胶、20μgSTAg+10μg蜂胶、20μgSTAg+20μg蜂胶及20μgSTAg+40μg蜂胶鼻内免疫小鼠2次,间隔14d。末次免疫后第10天,用RH株弓形虫速殖子灌胃攻击,4×104个/只,观察小鼠存活情况。攻击后第30天处死全部小鼠,分离计数小鼠脑、肝组织内速殖子和肠系膜淋巴结淋巴细胞(MLNL)、脾T淋巴细胞;ELISA法检测小鼠肺冲洗液、粪便弓形虫特异性sIgA和血清IgG抗体水平。结果随着蜂胶剂量的增加,小鼠脑、肝速殖子数呈下降趋势,20μgSTAg+40μg蜂胶组显著低于20μgSTAg组,脑、肝组织减虫率分别为46.10%和60.11%;MLNL和脾T淋巴细胞增殖活性提高,其中20μgSTAg+40μg蜂胶组小鼠MLNL和脾T淋巴细胞数分别为(2.09±0.24)×107和(1.89±0.21)×107,均显著高于20μgSTAg组;肺冲洗液、粪便弓形虫特异性sIgA和血清IgG抗体水平呈增高趋势,但各组间差异无统计学意义。结论STAg联合蜂胶佐剂鼻内免疫小鼠可有效诱导抗弓形虫感染的保护作用,适宜的蜂胶剂量为40μg。
Objective To investigate the protective effect of intranasal immunization with soluble tachyzoite antigen (STAg) combined with propolis adjuvant against Toxoplasma gondii infection and the appropriate dosage of propolis. Methods Sixty BALB / c mice were randomly divided into 5 groups with 12 rats in each group. Mice were immunized intranasally with 20μgSTAg, 20μgSTAg + 5μg propolis, 20μgSTAg + 10μg propolis, 20μgSTAg + 20μg propolis and 20μgSTAg + 40μg propolis respectively , Interval 14d. On the 10th day after the last immunization, Toxoplasma gondii T.gondii gonorrhoeae were intragastrically challenged with 4 × 10 4 cells / mouse to observe the survival of the mice. All mice were sacrificed on the 30th day after challenge, and the counts of tachyzoites and mesenteric lymph node lymphocytes (MLNL) and splenic T lymphocytes were detected in the brain of mice, liver tissue, lung rinse and Toxoplasma gondii sIgA and serum IgG antibody levels. Results With the increase of propolis dose, the numbers of brain and liver tachyzoites decreased in 20μgSTAg + 40μg propolis group and 20μgSTAg group. The worm reduction rates in brain and liver tissues were 46.10% and 60.11%, respectively. MLNL and spleen T lymphocyte proliferation activity was enhanced. The numbers of MLNL and splenic T lymphocytes in 20μgSTAg + 40μg propolis group were (2.09 ± 0.24) × 107 and (1.89 ± 0.21) × 107, respectively, which were significantly higher than that of 20μgSTAg group. , Toxoplasma gondii specific sIgA and serum IgG antibody levels showed an upward trend, but no significant difference between the groups. Conclusion STAg combined with propolis adjuvant intranasally immunized mice can effectively protect against Toxoplasma gondii infection, the appropriate propolis dose of 40μg.