T-cell libraries allow simple parallel generation of multiple peptide-specific human T-cell clones.
J Immunol Methods
; 430: 43-50, 2016 Mar.
Article
em En
| MEDLINE
| ID: mdl-26826277
ABSTRACT
Isolation of peptide-specific T-cell clones is highly desirable for determining the role of T-cells in human disease, as well as for the development of therapies and diagnostics. However, generation of monoclonal T-cells with the required specificity is challenging and time-consuming. Here we describe a library-based strategy for the simple parallel detection and isolation of multiple peptide-specific human T-cell clones from CD8(+) or CD4(+) polyclonal T-cell populations. T-cells were first amplified by CD3/CD28 microbeads in a 96U-well library format, prior to screening for desired peptide recognition. T-cells from peptide-reactive wells were then subjected to cytokine-mediated enrichment followed by single-cell cloning, with the entire process from sample to validated clone taking as little as 6 weeks. Overall, T-cell libraries represent an efficient and relatively rapid tool for the generation of peptide-specific T-cell clones, with applications shown here in infectious disease (Epstein-Barr virus, influenza A, and Ebola virus), autoimmunity (type 1 diabetes) and cancer.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Peptídeos
/
Linfócitos T CD4-Positivos
/
Linfócitos T CD8-Positivos
Limite:
Humans
Idioma:
En
Revista:
J Immunol Methods
Ano de publicação:
2016
Tipo de documento:
Article
País de afiliação:
Reino Unido