title: Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA.
abstract: OBJECTIVE
  Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association.
  
  
  METHODS
  Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR.
  
  
  RESULTS
  The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10(-4), OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10(-7), OR 0.71; case-only pmeta=1.9×10(-4), OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR.
  
  
  CONCLUSIONS
  These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.
authors.affiliation: Division of Rheumatology, Department of Medicine, University of California at Los Angeles, Los Angeles, California, USA.
authors.forename: Jian
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Division of Rheumatology, University of Colorado School of Medicine, Aurora, Colorado, USA.
authors.forename: Brendan M
authors.initials: BM
authors.lastname: Giles
authors.affiliation: School of Pathology and Laboratory Medicine, Centre for Genetic Origins of Health and Disease, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Rhonda L
authors.initials: RL
authors.lastname: Taylor
authors.affiliation: Division of Rheumatology, University of Colorado School of Medicine, Aurora, Colorado, USA.
authors.forename: Gabriel A
authors.initials: GA
authors.lastname: Yette
authors.affiliation: Division of Rheumatology, University of Colorado School of Medicine, Aurora, Colorado, USA.
authors.forename: Kara M
authors.initials: KM
authors.lastname: Lough
authors.affiliation: School of Pathology and Laboratory Medicine, Centre for Genetic Origins of Health and Disease, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Han Leng
authors.initials: HL
authors.lastname: Ng
authors.affiliation: School of Pathology and Laboratory Medicine, Centre for Genetic Origins of Health and Disease, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Abraham
authors.affiliation: Division of Rheumatology, Department of Medicine, University of California at Los Angeles, Los Angeles, California, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Kelly
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Stuart B
authors.initials: SB
authors.lastname: Glenn
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Adler
authors.affiliation: Department of Biostatistical Sciences and Center for Public Health Genomics, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Adrienne H
authors.initials: AH
authors.lastname: Williams
authors.affiliation: Department of Biostatistical Sciences and Center for Public Health Genomics, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Mary E
authors.initials: ME
authors.lastname: Comeau
authors.affiliation: Department of Biostatistical Sciences and Center for Public Health Genomics, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Julie T
authors.initials: JT
authors.lastname: Ziegler
authors.affiliation: Department of Biostatistical Sciences and Center for Public Health Genomics, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Miranda
authors.initials: M
authors.lastname: Marion
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA Pfizer-Universidad de Granada-Junta de Andalucía Center for Genomics and Oncological Research, Granada, Spain.
authors.forename: Marta E
authors.initials: ME
authors.lastname: Alarcón-Riquelme
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Graciela S
authors.initials: GS
authors.lastname: Alarcón
authors.affiliation: Center for Autoimmune Diseases Research (CREA), Universidad del Rosario, Bogotá, Colombia.
authors.forename: Juan-Manuel
authors.initials: JM
authors.lastname: Anaya
authors.affiliation: Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul, South Korea.
authors.forename: Sang-Cheol
authors.initials: SC
authors.lastname: Bae
authors.affiliation: Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul, South Korea.
authors.forename: Dam
authors.initials: D
authors.lastname: Kim
authors.affiliation: Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul, South Korea.
authors.forename: Hye-Soon
authors.initials: HS
authors.lastname: Lee
authors.affiliation: Rosalind Russell/Ephraim P. Engleman Rheumatology Research Center, University of California San Francisco, San Francisco, California, USA.
authors.forename: Lindsey A
authors.initials: LA
authors.lastname: Criswell
authors.affiliation: Department of Internal Medicine, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Barry I
authors.initials: BI
authors.lastname: Freedman
authors.affiliation: Division of Rheumatology, Medical University of South Carolina, Charleston, South Carolina, USA.
authors.forename: Gary S
authors.initials: GS
authors.lastname: Gilkeson
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Joel M
authors.initials: JM
authors.lastname: Guthridge
authors.affiliation: Department of Medicine, University of Southern California, Los Angeles, California, USA.
authors.forename: Chaim O
authors.initials: CO
authors.lastname: Jacob
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA.
authors.forename: Judith A
authors.initials: JA
authors.lastname: James
authors.affiliation: Division of Rheumatology, Medical University of South Carolina, Charleston, South Carolina, USA.
authors.forename: Diane L
authors.initials: DL
authors.lastname: Kamen
authors.affiliation: Department of Clinical Pharmacology, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Joan T
authors.initials: JT
authors.lastname: Merrill
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA.
authors.forename: Kathy Moser
authors.initials: KM
authors.lastname: Sivils
authors.affiliation: Division of Rheumatology and Department of Immunology, Mayo Clinic, Rochester, Minnesota, USA.
authors.forename: Timothy B
authors.initials: TB
authors.lastname: Niewold
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Petri
authors.affiliation: Division of Rheumatology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
authors.forename: Rosalind
authors.initials: R
authors.lastname: Ramsey-Goldman
authors.affiliation: Department of Rheumatology and Clinical Immunogenetics, University of Texas Health Science Center at Houston, Houston, Texas, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Reveille
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA US Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma, USA.
authors.forename: R Hal
authors.initials: RH
authors.lastname: Scofield
authors.affiliation: Division of Rheumatology, Department of Pediatrics, University of Washington, Seattle, Washington, USA Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Stevens
authors.affiliation: Division of Rheumatology, Department of Medicine, University of Puerto Rico Medical Sciences Campus, San Juan, Puerto Rico.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Vilá
authors.affiliation: Division of Genetics and Molecular Medicine and Immunology, King's College London, London, UK.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Vyse
authors.affiliation: Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA US Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA.
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Kaufman
authors.affiliation: Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA US Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA.
authors.forename: John B
authors.initials: JB
authors.lastname: Harley
authors.affiliation: Department of Biostatistical Sciences and Center for Public Health Genomics, Wake Forest School of Medicine, Winston-Salem, North Carolina, USA.
authors.forename: Carl D
authors.initials: CD
authors.lastname: Langefeld
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Gaffney
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA Department of Epidemiology, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Brown
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Edberg
authors.affiliation: Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Kimberly
authors.affiliation: School of Pathology and Laboratory Medicine, Centre for Genetic Origins of Health and Disease, The University of Western Australia, Crawley, Western Australia, Australia.
authors.forename: Daniela
authors.initials: D
authors.lastname: Ulgiati
authors.affiliation: Division of Rheumatology, Department of Medicine, University of California at Los Angeles, Los Angeles, California, USA.
authors.forename: Betty P
authors.initials: BP
authors.lastname: Tsao
authors.affiliation: Division of Rheumatology, University of Colorado School of Medicine, Aurora, Colorado, USA Denver Veterans Affairs Medical Center, Denver, Colorado, USA.
authors.forename: Susan A
authors.initials: SA
authors.lastname: Boackle
mesh_terms: D000293:Adolescent; D000328:Adult; D000974:Antibodies, Antinuclear / Q000097:blood*; D016175:B-Lymphocyte Subsets / Q000276:immunology; D016022:Case-Control Studies; D004247:DNA / Q000276:immunology; D020022:Genetic Predisposition to Disease; D014644:Genetic Variation; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology; D008875:Middle Aged; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D017463:Receptors, Complement 3b / Q000096:biosynthesis; D017464:Receptors, Complement 3d / Q000235:genetics*; D018570:Risk Assessment / Q000379:methods; D014157:Transcription Factors / Q000378:metabolism; D055815:Young Adult
chemical_list: D000974:Antibodies, Antinuclear; C509991:CR1 protein, human; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d; D014157:Transcription Factors; D004247:DNA
keywords: Autoantibodies; B cells; Gene Polymorphism; Systemic Lupus Erythematosus