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目的观察不同剂量重组弓形虫Peroxiredoxin蛋白(rTgPrx)诱导小鼠的体液免疫、细胞免疫应答及其抗弓形虫感染的保护作用。方法BALB/c小鼠60只,随机分为5组,分别用20、40、60、80μgrTgPrx(溶于100μlPBS中)和100μlPBS皮下免疫小鼠3次,间隔2周。末次免疫后第14天,用1×104个速殖子/只灌胃攻击,逐日观察小鼠健康状况。攻击后第30天,ELISA法检测小鼠血清IgG和小肠冲洗液sIgA水平,分离并计数脾淋巴细胞和小肠上皮内淋巴细胞(IEL),分离肝、脑组织内弓形虫速殖子并计数。结果60μgrTgPrx组小鼠血清IgG水平显著高于PBS、20μg和40μgrTgPrx组,小肠冲洗液sIgA组间比较差异无统计学意义;80μgrTgPrx组小鼠脾淋巴细胞数量明显高于PBS和20μgrTgPrx组,IEL数量各组间差异无统计学意义;60μg和80μgrTgPrx组小鼠脑和肝组织虫荷均低于PBS和20μgrTgPrx组。结论60μg和80μgrTgPrx皮下免疫小鼠可诱导有效的体液免疫、细胞免疫应答,且抗弓形虫感染的保护作用效果良好。
Objective To observe the humoral and cellular immune responses induced by Peroxiredoxin protein (rTgPrx) in different doses of recombinant Toxoplasma gondii and its protective effect against Toxoplasma gondii infection. Methods Sixty BALB / c mice were randomly divided into 5 groups. The mice were immunized subcutaneously with 20, 40, 60 and 80 μg rTgPrx (100 μl PBS) and 100 μl PBS for 3 weeks respectively. On the 14th day after the last immunization, mice were challenged daily with 1 × 10 4 tachyzoites / gavage challenge. Thirty days after the challenge, serum IgG and intestinal sIgA levels were measured by ELISA. Splenic lymphocytes and small intestine intraepithelial lymphocytes (IELs) were isolated and counted. Toxoplasma gondii were isolated from liver and brain and counted. Results The serum IgG levels in 60μg rTgPrx group were significantly higher than those in PBS group, 20μg and 40μg rTgPrx group, but there was no significant difference in intestinal rinse sIgA group. The number of splenic lymphocytes in 80μg rTgPrx group was significantly higher than that in PBS group and 20μg rTgPrx group There was no significant difference between the two groups. The mouse and liver tissue biotypes in the 60 μg and 80 μg rTgPrx groups were lower than those in the PBS and 20 μg rTgPrx groups. Conclusion The mice immunized subcutaneously with 60μg and 80μg rTgPrx can induce effective humoral and cellular immune responses, and the protective effect against Toxoplasma gondii infection is good.