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目的观察弓形虫速殖子排泄-分泌抗原(excreted/secreted antigen,ESA)和可溶性速殖子抗原(soluble tachyzoite antigen,STAg)鼻内免疫小鼠的免疫原性。方法BALB/c小鼠随机分为4组,分别用PBS 20μl/只、体外排泄-分泌抗原(excreted/secreted antigenin vitro,ESAv)、腹腔排泄-分泌抗原(excreted/secreted antigen in mice,ESAm)和STAg各20μg/只鼻内免疫2次,间隔14 d。分别于末次免疫后14 d和44 d每组处死8只小鼠,计数肠上皮内淋巴细胞(intestinal intraepithelial lymphocytes,iIEL)和脾淋巴细胞,ELISA法检测血清IgG和小肠冲洗液sIgA抗体水平。结果实验期间,ESAm组小鼠于二次免疫后状态欠佳,其他各组小鼠健康状况良好。末次免疫后14 d,各抗原组脾淋巴细胞及iIEL均增殖活跃,细胞数与PBS组比较,差异具统计学意义(P<0.05或P<0.01);至免疫后44 d,两种ESA组脾淋巴细胞及iIEL数与PBS组比较差异具统计学意义(P<0.05)。各抗原组血清IgG水平在免疫后14 d和44 d均明显增高,与PBS组比较差异有统计学意义(P<0.05或P<0.01)。免疫后14 d肠液sIgA水平ESAv、ES-Am和STAg组与PBS组比较差异有统计学意义(P<0.05或P<0.01),ESAm和STAg组与ESAv组比较差异有统计学意义(P<0.05或P<0.01),两种ESA组在免疫后44 d与PBS组比较差异仍具统计学意义(P<0.05)。结论ESAv、ESAm和STAg鼻内免疫均可诱导粘膜及系统的细胞和体液免疫应答,有较强的免疫原性。但ESAm可能对机体有毒副作用,不适宜直接鼻内免疫。
Objective To observe the immunogenicity of mice immunized intranasally with Toxoplasma gondii excreted / secreted antigen (ESA) and soluble tachyzoite antigen (STAg). Methods BALB / c mice were randomly divided into 4 groups. The mice were immunized with PBS 20μl / excreted / secreted antigen in vitro (ESAv), excreted / secreted antigen in mice (ESAm) and STAg each 20μg / only intranasal immunization 2 times, an interval of 14 d. Eight mice were sacrificed on the 14th day and the 44th day after the last immunization, respectively. The intestinal intraepithelial lymphocytes (iIEL) and splenic lymphocytes were counted. Serum IgG and sIgA antibody levels were measured by ELISA. Results During the experiment, mice in ESAm group were not in good condition after secondary immunization, and mice in other groups were in good health. At 14 days after the last immunization, splenic lymphocytes and iIEL in each antigen group proliferated actively. The number of cells was significantly different from PBS group (P <0.05 or P <0.01) The splenic lymphocytes and iIEL numbers were significantly different from PBS group (P <0.05). Serum IgG levels of each antigen group were significantly increased at 14 d and 44 d after immunization compared with PBS group (P <0.05 or P <0.01). The levels of sIgA in intestinal fluid of ESAv, ES-Am and STAg groups at 14th day after immunization were significantly different from those in PBS group (P <0.05 or P <0.01). There was significant difference between ESAm group and STAg group and ESAv group (P < 0.05 or P <0.01). The difference between the two ESA groups at 44 d after immunization and PBS group was still statistically significant (P <0.05). Conclusion Both intranasal immunization with ESAv, ESAm and STAg can induce cellular and humoral immune responses in mucosa and systemic and have strong immunogenicity. But ESAm may have toxic side effects on the body, not suitable for direct intranasal immunization.