Harmonisation of short-term in vitro culture for the expansion of antigen-specific CD8(+) T cells with detection by ELISPOT and HLA-multimer staining.

Détails

Ressource 1Télécharger: BIB_DCEBADBFBEE5.P001.pdf (1945.44 [Ko])
Etat: Public
Version: de l'auteur⸱e
ID Serval
serval:BIB_DCEBADBFBEE5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Harmonisation of short-term in vitro culture for the expansion of antigen-specific CD8(+) T cells with detection by ELISPOT and HLA-multimer staining.
Périodique
Cancer Immunology, Immunotherapy : Cii
Auteur⸱e⸱s
Chudley L., McCann K.J., Coleman A., Cazaly A.M., Bidmon N., Britten C.M., van der Burg S.H., Gouttefangeas C., Jandus C., Laske K., Maurer D., Romero P., Schröder H., Stynenbosch L.F., Walter S., Welters M.J., Ottensmeier C.H.
ISSN
1432-0851 (Electronic)
ISSN-L
0340-7004
Statut éditorial
Publié
Date de publication
2014
Volume
63
Numéro
11
Pages
1199-211
Langue
anglais
Notes
Publication types: JOURNAL ARTICLE
Résumé
Ex vivo ELISPOT and multimer staining are well-established tests for the assessment of antigen-specific T cells. Many laboratories are now using a period of in vitro stimulation (IVS) to enhance detection. Here, we report the findings of a multi-centre panel organised by the Association for Cancer Immunotherapy Immunoguiding Program to investigate the impact of IVS protocols on the detection of antigen-specific T cells of varying ex vivo frequency. Five centres performed ELISPOT and multimer staining on centrally prepared PBMCs from 3 donors, both ex vivo and following IVS. A harmonised IVS protocol was designed based on the best-performing protocol(s), which was then evaluated in a second phase on 2 donors by 6 centres. All centres were able to reliably detect antigen-specific T cells of high/intermediate frequency both ex vivo (Phase I) and post-IVS (Phase I and II). The highest frequencies of antigen-specific T cells ex vivo were mirrored in the frequencies following IVS and in the detection rates. However, antigen-specific T cells of a low/undetectable frequency ex vivo were not reproducibly detected post-IVS. Harmonisation of the IVS protocol reduced the inter-laboratory variation observed for ELISPOT and multimer analyses by approximately 20 %. We further demonstrate that results from ELISPOT and multimer staining correlated after (P < 0.0001 and R (2) = 0.5113), but not before IVS. In summary, IVS was shown to be a reproducible method that benefitted from method harmonisation.
Pubmed
Web of science
Open Access
Oui
Création de la notice
29/09/2014 11:35
Dernière modification de la notice
20/08/2019 16:01
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