title: Human T cell clones reactive against U-small nuclear ribonucleoprotein autoantigens from connective tissue disease patients and healthy individuals.
abstract: SLE and mixed connective tissue disease (MCTD) are characterized by the presence of high titers of autoantibodies against uridine-rich RNA-small nuclear ribonucleoprotein (snRNP) Ag. Because the presence of such snRNP-reactive autoantibodies has recently been shown to be associated with polymorphisms of HLA, this study was undertaken to determine whether snRNP-reactive T cells could be identified and characterized from patients. PBMC were stimulated with affinity-purified snRNP Ag and cloned by limiting dilution in the presence of rIL-2 and rIL-4, snRNP-reactive human T cell clones were generated from three patients and two healthy blood donors who possessed disease-associated HLA genotypes. The cell surface phenotype of clones determined by flow cytometry was CD3+, CD4+, CD45RO+, TCR V alpha beta+. TCR V beta analysis, performed using V beta-specific primers and polymerase chain reaction, revealed that the T cell lines generated were clonal; a limited number of TCR V beta genes were expressed among the clones tested. All clones tested by mAb blocking of Ag-induced proliferation were restricted by HLA-DR. Several T cell clones were identified that were specific for B'/B or D polypeptides. These results demonstrate that snRNP-reactive T cells can be isolated from SLE and MCTD patients in vitro, and that Ag-driven expansion of such T cells could play a role in the immunopathogenesis of these diseases in vivo.
authors.affiliation: Department of Internal Medicine, University of Missouri-Columbia School of Medicine.
authors.forename: R W
authors.initials: RW
authors.lastname: Hoffman
authors.forename: Y
authors.initials: Y
authors.lastname: Takeda
authors.forename: G C
authors.initials: GC
authors.lastname: Sharp
authors.forename: D R
authors.initials: DR
authors.lastname: Lee
authors.forename: D L
authors.initials: DL
authors.lastname: Hill
authors.forename: H
authors.initials: H
authors.lastname: Kaneoka
authors.forename: C W
authors.initials: CW
authors.lastname: Caldwell
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D002999:Clone Cells; D005260:Female; D006684:HLA-DR Antigens / Q000502:physiology; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D008947:Mixed Connective Tissue Disease / Q000276:immunology*; D008969:Molecular Sequence Data; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis; D017411:Ribonucleoproteins, Small Nuclear / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D006684:HLA-DR Antigens; D016693:Receptors, Antigen, T-Cell, alpha-beta; D017411:Ribonucleoproteins, Small Nuclear