title: Lack of T cell oligoclonality in enzyme-digested synovial tissue and in synovial fluid in most patients with rheumatoid arthritis.
abstract: The dominant presence of specific T-cell populations in the rheumatoid joint as detected by Southern blot analysis of T cell receptor (TCR) gene rearrangements would indicate local antigen recognition and T cell proliferation. We therefore studied TCR beta chain gene rearrangements using a C beta 2 probe in paired samples of T cell populations from synovial tissue and peripheral blood (n = 6) as well as synovial fluid (n = 16) and peripheral blood (n = 18) of patients with rheumatoid arthritis (RA). Peripheral blood mononuclear cells from healthy donors (n = 7) served as a control. T cells were studied directly after isolation or after non-specific expansion with OKT3 monoclonal antibody (MoAb) and T cell growth factor (TCGF). DNA samples were digested with EcoRI and HindIII to detect rearrangements to C beta 1 and C beta 2, respectively. Extra bands were detected in all EcoRI-digested DNA samples prepared from both freshly isolated and non-specifically expanded T cell populations of patients and healthy donors, possibly representing 'common' (V-) D-J rearrangements. Dominant rearrangements were found in only two out of 16 synovial fluid T cell populations (one freshly isolated and one expanded) and not in peripheral blood or synovial tissue derived T cell populations. No extra bands were detected in HindIII-digested DNA samples. To investigate the effect of in vitro culture techniques on rearrangement patterns we studied DNA samples prepared from synovial tissue T cells obtained both by outgrowth from tissue with TCGF or by enzyme digestion and subsequent expansion either with TCGF or with OKT3 MoAb and TCGF. Whereas the latter T cell population yielded 'common' rearrangements, the former T cell populations yielded different dominant rearrangements. These data indicate that oligoclonality of the T cell populations in synovial tissue and synovial fluid of patients with RA is a rare event. The data also show the influence of in vitro culture techniques on the result of TCR gene rearrangement analysis.
authors.affiliation: Department of Rheumatology, University Hospital, Leiden, The Netherlands.
authors.forename: J M
authors.initials: JM
authors.lastname: Van Laar
authors.forename: A M
authors.initials: AM
authors.lastname: Miltenburg
authors.forename: M J
authors.initials: MJ
authors.lastname: Verdonk
authors.forename: M R
authors.initials: MR
authors.lastname: Daha
authors.forename: R R
authors.initials: RR
authors.lastname: De Vries
authors.forename: P J
authors.initials: PJ
authors.lastname: Van den Elsen
authors.forename: F C
authors.initials: FC
authors.lastname: Breedveld
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D015139:Blotting, Southern; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D005260:Female; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D006801:Humans; D006821:Hyaluronoglucosaminidase; D007376:Interleukin-2 / Q000502:physiology; D008297:Male; D003012:Microbial Collagenase; D008875:Middle Aged; D016853:Muromonab-CD3; D013582:Synovial Fluid / Q000276:immunology*; D013583:Synovial Membrane / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D016853:Muromonab-CD3; D006821:Hyaluronoglucosaminidase; D003012:Microbial Collagenase