{Reference Type}: Journal Article
{Title}: Improved ELISPOT protocol for monitoring Th1/Th17 T-cell response following T.gondii infection.
{Author}: Fasquelle F;Vreulx AC;Betbeder D;
{Journal}: PLoS One
{Volume}: 19
{Issue}: 5
{Year}: 2024
{Factor}: 3.752
{DOI}: 10.1371/journal.pone.0301687
{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.