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  Analysis of the Paired TCR alpha- and beta-chains of Single Human T Cells

Kim, S.-M., Bhonsle, L., Besgen, P., Nickel, J., Backes, A., Held, K., et al. (2012). Analysis of the Paired TCR alpha- and beta-chains of Single Human T Cells. PLOS ONE, 7(5): e37338. doi:10.1371/journal.pone.0037338.

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Datensatz-Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-C3BA-1 Versions-Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-C3BB-0
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 Urheber:
Kim, Song-Min1, Autor
Bhonsle, Latika2, Autor              
Besgen, Petra1, Autor
Nickel, Jens1, Autor
Backes, Anna2, Autor              
Held, Kathrin1, Autor
Vollmer, Sigrid1, Autor
Dornmair, Klaus2, Autor              
Prinz, Joerg C.1, Autor
Affiliations:
1External Organizations, ou_persistent22              
2Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society, ou_1113547              

Inhalt

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Schlagwörter: MULTIPLE-SCLEROSIS LESIONS; CENTRAL-NERVOUS-SYSTEM; PSORIASIS-VULGARIS; CLONAL EXPANSIONS; SKIN-LESIONS; EXPRESSION; REARRANGEMENTS; REPERTOIRE; PERSIST; GENES
 Zusammenfassung: Analysis of the paired i.e. matching TCR alpha- and beta-chain rearrangements of single human T cells is required for a precise investigation of clonal diversity, tissue distribution and specificity of protective and pathologic T-cell mediated immune responses. Here we describe a multiplex RT-PCR based technology, which for the first time allows for an unbiased analysis of the complete sequences of both alpha- and beta-chains of TCR from single T cells. We validated our technology by the analysis of the pathologic T-cell infiltrates from tissue lesions of two T-cell mediated autoimmune diseases, psoriasis vulgaris (PV) and multiple sclerosis (MS). In both disorders we could detect various T cell clones as defined by multiple T cells with identical alpha- and beta-chain rearrangements distributed across the tissue lesions. In PV, single cell TCR analysis of lesional T cells identified clonal CD8(+) T cell expansions that predominated in the epidermis of psoriatic plaques. An MS brain lesion contained two dominant CD8(+) T-cell clones that extended over the white and grey matter and meninges. In both diseases several clonally expanded T cells carried dual TCRs composed of one V beta and two different V alpha-chain rearrangements. These results show that our technology is an efficient instrument to analyse alpha beta-T cell responses with single cell resolution in man. It should facilitate essential new insights into the mechanisms of protective and pathologic immunity in many human T-cell mediated conditions and allow for resurrecting functional TCRs from any alpha beta-T cell of choice that can be used for investigating their specificity.

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Sprache(n): eng - Englisch
 Datum: 2012-05-23
 Publikationsstatus: Online publiziert
 Seiten: 12
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: ISI: 000305335800037
DOI: 10.1371/journal.pone.0037338
 Art des Abschluß: -

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Titel: PLOS ONE
Genre der Quelle: Zeitschrift
 Urheber:
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Ort, Verlag, Ausgabe: 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA : PUBLIC LIBRARY SCIENCE
Seiten: - Band / Heft: 7 (5) Artikelnummer: e37338 Start- / Endseite: - Identifikator: ISSN: 1932-6203