Improved ELISPOT protocol for monitoring Th1/Th17 T-cell response following T.gondii infection.
PLoS One
; 19(5): e0301687, 2024.
Article
em En
| MEDLINE
| ID: mdl-38718078
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Toxoplasma
/
Toxoplasmose
/
Células Th1
/
Células Th17
/
ELISPOT
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
França