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目的研究分离自弓形虫RH株速殖子经口感染小鼠后不同时间点的小肠上皮内淋巴细胞(intraepitheliallymphocyte,IEL)过继转移抗弓形虫感染作用,探讨其作用机制。方法BALB/c鼠经口感染弓形虫速殖子5×104个/只或未感染,作为供体提供IEL。同品系受体鼠分为实验组(IEL7组、IEL9组、IEL11组、IEL13组和IEL15组)和对照组(IEL0组),每组6只小鼠。受体鼠经尾静脉分别接受分离自供体鼠经速殖子感染后第7、9、11、13、15天的致敏IEL或未致敏的IEL3×105/0.2ml只。各组小鼠过继转移后第4天,用弓形虫速殖子灌胃攻击,攻击后第13天分离纯化脑、肺、脾组织弓形虫速殖子并计数,测定肠液IgA含量。结果致敏IEL过继免疫可使受体鼠脑、肺、脾组织内弓形虫速殖子数显著减少,接受感染后第13天IEL的小鼠组织内速殖子数减少最为显著(P<0.01)脑、肺和脾组织内速殖子数比对照组分别减少81.13%,58.43%和70.97%。致敏IEL过继转移使肠道IgA水平升高,IEL11组和IEL13组显著高于IEL0组(P<0.01)。结论致敏IEL过继转移能上调肠道黏膜的免疫应答,导致黏膜特异性IgA分泌增加,诱导黏膜抗体的保护性免疫应答。IEL的这种保护作用具有致敏的时间依赖性,感染后第13天分离的致敏IEL对弓形虫感染具有最大的保护作用。
Objective To study the effect of intraperitoneal lymphocyte (IEL) adoptive transfer of Toxoplasma gondii isolated from mice infected with Tachyzoites of Toxoplasma gondii RH strain at different time points to investigate their mechanism. Methods BALB / c mice were orally infected with Toxoplasma gondii tachyzoites at 5 × 10 4 / animal or uninfected, providing IEL as a donor. The same strain of recipient mice were divided into experimental group (IEL7 group, IEL9 group, IEL11 group, IEL13 group and IEL15 group) and control group (IEL0 group), 6 mice in each group. The recipient mice received sensitized IEL on the 7th, 9th, 11th, 13th, and 15th day after the tachyzoite infection from the donor rat via the tail vein, respectively, or non-sensitized IEL3x105 / 0.2ml only. On the 4th day after adoptive transfer, the tachyzoites of Toxoplasma gondii were challenged with gonadotrophs. Toxoplasma gondii of brain, lung and spleen were isolated and purified on the 13th day after challenge, and the content of IgA in intestinal juice was determined. Results The sensitized IEL adoptive immunization could significantly reduce Toxoplasma gondii tachyzoites in the brain, lung and spleen of recipient mice. The reduction of tachyzoites in IEL mice was the most significant on the 13th day after infection (P <0.01 ) Tachyzoites in the brain, lung and spleen decreased by 81.13%, 58.43% and 70.97% respectively compared with the control group. IEL11 and IEL13 groups were significantly higher than IEL0 group (P <0.01). Conclusion The adoptive transfer of sensitized IEL can upregulate the intestinal mucosal immune response, leading to increased mucosal-specific IgA secretion and inducing a protective immune response to mucosal antibodies. This protective effect of IEL is sensitized in a time-dependent manner, with the sensitized IEL isolated on day 13 post infection having the greatest protective effect against Toxoplasma gondii infection.