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  Clonal tracking of autoaggressive T cells in polymyositis by combining laser microdissection, single-cell PCR, and CDR3-spectratype analysis

Hofbauer, M., Wiesener, S., Babbe, H., Roers, A., Wekerle, H., Dornmair, K., et al. (2003). Clonal tracking of autoaggressive T cells in polymyositis by combining laser microdissection, single-cell PCR, and CDR3-spectratype analysis. Proceedings of the National Academy of Sciences U S A, 100(7), 4090-4095.

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 Creators:
Hofbauer, Monika1, Author              
Wiesener, S.2, Author
Babbe, H.2, Author
Roers, A.2, Author
Wekerle, Hartmut1, Author              
Dornmair, Klaus1, Author              
Goebels, Norbert1, Author              
Affiliations:
1Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society, ou_1113547              
2Institute for Clinical Neuroimmunology, Ludwig Maximilians University, D-81377 Munich, Germany; Department of Neuroimmunology, Max Planck Institute for Neurobiology, 82152 Martinsried, Germany; and Institute for Genetics and Department of Dermatology, University of Cologne, 50931 Cologne, Germany, ou_persistent22              

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 Abstract: Clonal expansions of CD8+ T cells have been identified in muscle and blood of polymyositis patients by PCR techniques, including T cell receptor (TCR) complementarity-determining region (CDR)3 length analysis (spectratyping). To examine a possible pathogenic role of these clonally expanded T cells, we combined CDR3 spectratyping with laser microdissection and single-cell PCR of individual myocytotoxic T cells that contact, invade, and destroy a skeletal muscle fiber. First, we screened cDNA from muscle biopsy specimens by CDR3 spectratyping for expanded TCR chain variable region (BV) sequences. To pinpoint the corresponding T cells in tissue, we stained cryostat sections with appropriate anti-TCR BV mAbs, isolated single BV+ T cells that directly contacted or invaded a muscle fiber by laser-assisted microdissection, and amplified their TCR BV chain sequences from rearranged genomic DNA. In this way, we could relate the oligoclonal peaks identified by CDR3-spectratype screening to morphologically characterized microdissected T cells. In one patient, a large fraction of the microdissected T cells carried a common TCR-BV amino acid CDR3 motif and conservative nucleotide exchanges in the CDR3 region, suggesting an antigen-driven response. In several cases, we tracked these T cell clones for several years in CD8+ (but not CD4+) blood lymphocytes and in two patients also in consecutive muscle biopsy specimens. During immunosuppressive therapy, oligoclonal CDR3-spectratype patterns tended to revert to more polyclonal Gaussian distribution-like patterns. Our findings demonstrate that CDR3 spectratyping and single-cell analysis can be combined to identify and track autoaggressive T cell clones in blood and target tissue. This approach should be applicable to other inflammatory and autoimmune disorders.

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Language(s): eng - English
 Dates: 2003-04-01
 Publication Status: Published in print
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 Identifiers: eDoc: 49900
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Title: Proceedings of the National Academy of Sciences U S A
  Alternative Title : PNAS
Source Genre: Journal
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Pages: - Volume / Issue: 100 (7) Sequence Number: - Start / End Page: 4090 - 4095 Identifier: -