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First, we screened cDNA from muscle biopsy specimens by CDR3 spectratyping for expanded TCR \u03b2 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.<\/jats:p>","DOI":"10.1073\/pnas.0236183100","type":"journal-article","created":{"date-parts":[[2003,4,1]],"date-time":"2003-04-01T14:25:08Z","timestamp":1049207108000},"page":"4090-4095","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":88,"title":["Clonal tracking of autoaggressive T cells in polymyositis by combining laser microdissection, single-cell PCR, and CDR3-spectratype analysis"],"prefix":"10.1073","volume":"100","author":[{"given":"Monika","family":"Hofbauer","sequence":"first","affiliation":[{"name":"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, 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