Measuring T cell receptor and T cell gene expression diversity in antigen-responsive human CD4+ T cells.
J Immunol Methods
; 400-401: 13-22, 2013 Dec 31.
Article
em En
| MEDLINE
| ID: mdl-24239865
ABSTRACT
T cells have diversity in TCR, epitope recognition, and cytokine production, and can be used for immune monitoring. Furthermore, clonal expansion of TCR families in disease may provide opportunities for TCR-directed therapies. We developed methodology for sequencing expressed genes of TCR alpha and beta chains from single cells and applied this to vaccine (tetanus-toxoid)-responsive CD4(+) T cells. TCR alpha and beta chains were both successfully sequenced in 1309 (43%) of 3038 CD4(+) T cells yielding 677 different receptors. TRAV and TRBV gene usage differed between tetanus-toxoid-responsive and non-responsive cells (p=0.004 and 0.0002), and there was extensive TCR diversity in tetanus-toxoid-responsive cells within individuals. Identical TCRs could be recovered in different samples from the same subject TCRs identified after booster vaccination were frequent in pre-booster memory T cells (31% of pre-booster TCR), and also identified in pre-booster vaccination naïve cells (6.5%). No TCR was shared between subjects, but tetanus toxoid-responsive cells sharing one of their TCR chains were observed within and between subjects. Coupling single-cell gene expression profiling to TCR sequencing revealed examples of distinct cytokine profiles in cells bearing identical TCR. Novel molecular methodology demonstrates extensive diversity of Ag-responsive CD4(+) T cells within and between individuals.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Toxoide Tetânico
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Linfócitos T CD4-Positivos
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Subpopulações de Linfócitos T
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Receptores de Antígenos de Linfócitos T alfa-beta
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Análise de Sequência de DNA
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Perfilação da Expressão Gênica
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Análise de Célula Única
Limite:
Adult
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Female
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Humans
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Infant
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Male
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Middle aged
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article