{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T21:07:14Z","timestamp":1788469634341,"version":"build-2803163510"},"reference-count":27,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,8]],"date-time":"2024-05-08T00:00:00Z","timestamp":1715126400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In the monitoring of human <jats:italic>Toxoplasma gondii<\/jats:italic> 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 <jats:italic>T<\/jats:italic>.<jats:italic>gondii<\/jats:italic>-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-\u03b3, IL-2, IL-12 and IL-17 cytokines. This protocol allows to evaluate precisely the immune response after a <jats:italic>T<\/jats:italic>.<jats:italic>gondii<\/jats:italic> infection.<\/jats:p>","DOI":"10.1371\/journal.pone.0301687","type":"journal-article","created":{"date-parts":[[2024,5,8]],"date-time":"2024-05-08T17:37:14Z","timestamp":1715189834000},"page":"e0301687","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Improved ELISPOT protocol for monitoring Th1\/Th17 T-cell response following T.gondii infection"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9838-5006","authenticated-orcid":true,"given":"Francois","family":"Fasquelle","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana\u00efs-Camille","family":"Vreulx","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Didier","family":"Betbeder","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,5,8]]},"reference":[{"issue":"2\u20133","key":"pone.0301687.ref001","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.ijpara.2020.11.001","article-title":"Control of human toxoplasmosis","volume":"51","author":"N. 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Nanomedicine"},{"key":"pone.0301687.ref014","first-page":"14","article-title":"In silico and in vivo analysis of Toxoplasma gondii epitopes by correlating survival data with peptide\u2013MHC-I binding affinities","volume":"48","author":"Int","year":"2016","journal-title":"J. Infect. Dis."},{"issue":"1","key":"pone.0301687.ref015","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1038\/s41467-021-21856-3","article-title":"T cell assays differentiate clinical and subclinical SARS-CoV-2 infections from cross-reactive antiviral responses","volume":"12","author":"A. Ogbe","year":"2021","journal-title":"Nat. Commun."},{"issue":"4","key":"pone.0301687.ref016","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1007\/s11255-021-02960-1","article-title":"Value of gamma interferon enzyme-linked immunospot assay in the diagnosis of peritoneal dialysis-associated tuberculous peritonitis","volume":"54","author":"Q. Fan","year":"2022","journal-title":"Int. Urol. 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