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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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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-C3BA-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-000F-C3BB-0
Genre: Journal Article

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pone.0037338.pdf (Any fulltext), 461KB
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 Creators:
Kim, Song-Min1, Author
Bhonsle, Latika2, Author              
Besgen, Petra1, Author
Nickel, Jens1, Author
Backes, Anna2, Author              
Held, Kathrin1, Author
Vollmer, Sigrid1, Author
Dornmair, Klaus2, Author              
Prinz, Joerg C.1, Author
Affiliations:
1External Organizations, escidoc:persistent22              
2Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society, escidoc:1113547              

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Free keywords: MULTIPLE-SCLEROSIS LESIONS; CENTRAL-NERVOUS-SYSTEM; PSORIASIS-VULGARIS; CLONAL EXPANSIONS; SKIN-LESIONS; EXPRESSION; REARRANGEMENTS; REPERTOIRE; PERSIST; GENES
 Abstract: 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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Language(s): eng - English
 Dates: 2012-05-23
 Publication Status: Published online
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

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Title: PLOS ONE
Source Genre: Journal
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Publ. Info: 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA : PUBLIC LIBRARY SCIENCE
Pages: - Volume / Issue: 7 (5) Sequence Number: e37338 Start / End Page: - Identifier: ISSN: 1932-6203