Please note that eDoc will be permanently shut down in the first quarter of 2021!      Home News About Us Contact Contributors Disclaimer Privacy Policy Help FAQ

Quick Search
My eDoc
Session History
Support Wiki
Direct access to
document ID:

          Institute: MPI für Neurobiologie     Collection: Neuroimmunology     Display Documents

ID: 49900.0, MPI für Neurobiologie / Neuroimmunology
Clonal tracking of autoaggressive T cells in polymyositis by combining laser microdissection, single-cell PCR, and CDR3-spectratype analysis
Authors:Hofbauer, Monika; Wiesener, S.; Babbe, H.; Roers, A.; Wekerle, Hartmut; Dornmair, Klaus; Goebels, Norbert
Date of Publication (YYYY-MM-DD):2003-04-01
Title of Journal:Proceedings of the National Academy of Sciences U S A
Journal Abbrev.:PNAS
Issue / Number:7
Start Page:4090
End Page:4095
Review Status:not specified
Audience:Not Specified
Abstract / Description: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.
External Publication Status:published
Document Type:Article
Affiliations:MPI für Neurobiologie/Neuroimmunology (Wekerle)/Clinical Neuroimmunology (Hohlfeld)
External Affiliations:Institute 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
The scope and number of records on eDoc is subject to the collection policies defined by each institute - see "info" button in the collection browse view.