title: The T-cell-receptor repertoire in the synovial fluid of a patient with rheumatoid arthritis is polyclonal.
abstract: We have analyzed the T-cell-receptor repertoire expressed in the synovial fluid of a patient with rheumatoid arthritis by using an inverse polymerase chain reaction. Total RNA was isolated from Ficoll-purified mononuclear cells and converted into circularized double-stranded cDNA. Specific amplification of alpha- and beta-chain variable regions (V alpha and V beta) was achieved with inverted alpha- and beta-chain constant region (C alpha and C beta) primer pairs, and the amplification products were cloned into phage vectors. A total of 78 alpha and 76 beta clones were sequenced, and 67 and 72 productively rearranged alpha and beta genes were identified, respectively. Thirty-one V alpha, 33 alpha-chain joining region (J alpha), 29 V beta, and 12 beta-chain joining region (J beta) gene segments were found in the productively rearranged clones, indicating that the T-cell repertoire expressed in the synovial fluid of this RA patient is highly heterogenous and polyclonal. Comparison of peripheral blood and synovial fluid repertoires showed that the most abundant V beta sequences, V beta 2.1 and V beta 3.1, were enriched in the inflamed joint by a factor of 2 to 3. It is possible that T cells expressing these V beta gene segments, which recognize bacterial superantigens, play a role in the disease.
authors.affiliation: Pharmaceutical Research New Technologies, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
authors.forename: Y
authors.initials: Y
authors.lastname: Uematsu
authors.forename: H
authors.initials: H
authors.lastname: Wege
authors.forename: A
authors.initials: A
authors.lastname: Straus
authors.forename: M
authors.initials: M
authors.lastname: Ott
authors.forename: W
authors.initials: W
authors.lastname: Bannwarth
authors.forename: J
authors.initials: J
authors.lastname: Lanchbury
authors.forename: G
authors.initials: G
authors.lastname: Panayi
authors.forename: M
authors.initials: M
authors.lastname: Steinmetz
mesh_terms: D000595:Amino Acid Sequence; D001172:Arthritis, Rheumatoid / Q000276:immunology / Q000473:pathology*; D001483:Base Sequence; D002999:Clone Cells; D015332:Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics*; D013583:Synovial Membrane / Q000276:immunology; D013601:T-Lymphocytes / Q000473:pathology*
chemical_list: D009841:Oligonucleotides; D016693:Receptors, Antigen, T-Cell, alpha-beta