title: Functional assay of type I interferon in systemic lupus erythematosus plasma and association with anti-RNA binding protein autoantibodies.
abstract: OBJECTIVE
  Peripheral blood mononuclear cells (PBMCs) from patients with systemic lupus erythematosus (SLE) have increased expression of genes typically induced by type I interferon (IFN). However, it has been difficult to identify and quantify the factors responsible for activation of the IFN pathway in SLE. To characterize these mediators, we developed an assay that measures the functional effects of plasma or serum components on the gene expression of cultured target cells.
  
  
  METHODS
  WISH epithelial cell line cells were cultured with medium, with recombinant IFNalpha, IFNbeta, or IFNgamma, or with 50% plasma from SLE patients (n = 73), rheumatoid arthritis (RA) patients (n = 19), or healthy donors (n = 30). Real-time quantitative polymerase chain reaction was used to determine WISH cell expression of IFN target genes, including PRKR, IFIT1, IFI44, MX1, and C1orf29 (preferentially induced by IFNalpha) and CXCL9 (Mig) (preferentially induced by IFNgamma).
  
  
  RESULTS
  IFNalpha-regulated genes were induced by SLE plasma samples, but not by most of the RA or healthy control plasma samples. The activity in SLE plasma was inhibited >90% by anti-IFNalpha antibody, but not by anti-IFNbeta or anti-IFNgamma antibodies. The expression of each IFNalpha target gene induced by SLE plasma correlated with the expression of that gene studied ex vivo in PBMCs from the same patients and with the titer of anti-RNA binding protein (anti-RBP)-specific autoantibodies. Plasma activity paralleled PBMC expression of IFNalpha-inducible genes over time.
  
  
  CONCLUSIONS
  IFNalpha in SLE plasma is a major stimulus of IFN target gene expression and is related to expression of those genes in PBMCs from SLE patients and to the titer of anti-RBP autoantibodies. These data provide additional support for the view that IFNalpha mediates immune system activation and dysregulation in SLE.
authors.affiliation: Mary Kirkland Center for Lupus Research and Department of Medicine, Hospital for Special Surgery, and Weill Medical College of Cornell University, New York, New York 10021, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Hua
authors.forename: Kyriakos
authors.initials: K
authors.lastname: Kirou
authors.forename: Christina
authors.initials: C
authors.lastname: Lee
authors.forename: Mary K
authors.initials: MK
authors.lastname: Crow
mesh_terms: D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000235:genetics; D000943:Antigens, Differentiation; D001172:Arthritis, Rheumatoid / Q000097:blood; D001323:Autoantibodies; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D002460:Cell Line; D054370:Chemokine CXCL9; D019743:Chemokines, CXC / Q000235:genetics; D003598:Cytoskeletal Proteins / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007370:Interferon Type I / Q000097:blood*; D016898:Interferon-alpha / Q000276:immunology; D016899:Interferon-beta / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D007963:Leukocytes, Mononuclear; D008180:Lupus Erythematosus, Systemic / Q000097:blood* / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics; D062085:RNA, Long Noncoding; D016601:RNA-Binding Proteins / Q000235:genetics / Q000276:immunology*; D025521:Tumor Suppressor Proteins / Q000235:genetics
chemical_list: D000906:Antibodies; D000941:Antigens; D000943:Antigens, Differentiation; D001323:Autoantibodies; D051792:Basic Helix-Loop-Helix Proteins; D054370:Chemokine CXCL9; D019743:Chemokines, CXC; D003598:Cytoskeletal Proteins; D007370:Interferon Type I; D016898:Interferon-alpha; D016899:Interferon-beta; D007371:Interferon-gamma; D008565:Membrane Proteins; D011506:Proteins; D062085:RNA, Long Noncoding; D016601:RNA-Binding Proteins; D025521:Tumor Suppressor Proteins; C080810:CXCL9 protein, human; C114473:ERC2-IT1 lncRNA, human; C065888:IFI44 protein, human; C079440:MXI1 protein, human; C104647:PRKRA protein, human; C062010:leu-13 antigen