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Abstract：
In the monitoring of human Toxoplasma gondii infection, it is crucial to confirm the development of a specific Th1/Th17 immune response memory. The use of a simple, specific, and sensitive assay to follow the T-cell activation is thus required. Current protocols are not always specific as stimulation with peptides is Human Leukocyte Antigen (HLA)-dependent, while stimulation with total-lysis antigens tends to stimulate seronegative donors resulting to false positives. Here, an improved ELISPOT protocol is reported, using peripheral blood mononuclear cells (PBMC) of T.gondii-infected donors, incubated with the inactivated parasite. The results showed that, contrary to standard protocols, a pre-incubation step at high cell density in presence of the inactivated parasite allowed a specific Th1/Th17 response with the secretion of IFN-γ, IL-2, IL-12 and IL-17 cytokines. This protocol allows to evaluate precisely the immune response after a T.gondii infection.
摘要：
在人类弓形虫感染的监测中，确认特定Th1/Th17免疫反应记忆的发展至关重要。使用一个简单的，具体,因此需要灵敏的测定来跟踪T细胞活化。目前的方案并不总是特异性的，因为肽刺激是人类白细胞抗原（HLA）依赖性的，而用总裂解抗原刺激往往会刺激血清阴性供体，导致假阳性。这里,报告了一种改进的ELISPOT协议，使用T.gondii感染的供体的外周血单核细胞（PBMC），与灭活的寄生虫一起孵育。结果表明,与标准协议相反，在灭活的寄生虫存在下，高细胞密度的预孵育步骤允许分泌IFN-γ的特定Th1/Th17反应，IL-2、IL-12和IL-17细胞因子。该方案允许精确评估弓形虫感染后的免疫应答。
