Vaxinano, Loos, France.
PLoS One. 2024 May 8;19(5):e0301687. doi: 10.1371/journal.pone.0301687. eCollection 2024.
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.
在监测人类弓形虫感染时，确认特定的 Th1/Th17 免疫反应记忆的发展至关重要。因此，需要使用简单、特异和敏感的方法来跟踪 T 细胞的激活。目前的方案并不总是特异的，因为肽刺激依赖于人类白细胞抗原（HLA），而全裂解抗原的刺激往往会刺激血清阴性供体，导致假阳性。在这里，报告了一种改进的 ELISPOT 方案，使用弓形虫感染供体的外周血单核细胞（PBMC），用失活的寄生虫孵育。结果表明，与标准方案相反，在高细胞密度存在下用失活寄生虫进行预孵育步骤允许以 IFN-γ、IL-2、IL-12 和 IL-17 细胞因子的方式进行特异性 Th1/Th17 反应。该方案可精确评估弓形虫感染后的免疫反应。
