title: Violation of an evolutionarily conserved immunoglobulin diversity gene sequence preference promotes production of dsDNA-specific IgG antibodies.
abstract: Variability in the developing antibody repertoire is focused on the third complementarity determining region of the H chain (CDR-H3), which lies at the center of the antigen binding site where it often plays a decisive role in antigen binding. The power of VDJ recombination and N nucleotide addition has led to the common conception that the sequence of CDR-H3 is unrestricted in its variability and random in its composition. Under this view, the immune response is solely controlled by somatic positive and negative clonal selection mechanisms that act on individual B cells to promote production of protective antibodies and prevent the production of self-reactive antibodies. This concept of a repertoire of random antigen binding sites is inconsistent with the observation that diversity (DH) gene segment sequence content by reading frame (RF) is evolutionarily conserved, creating biases in the prevalence and distribution of individual amino acids in CDR-H3. For example, arginine, which is often found in the CDR-H3 of dsDNA binding autoantibodies, is under-represented in the commonly used DH RFs rearranged by deletion, but is a frequent component of rarely used inverted RF1 (iRF1), which is rearranged by inversion. To determine the effect of altering this germline bias in DH gene segment sequence on autoantibody production, we generated mice that by genetic manipulation are forced to utilize an iRF1 sequence encoding two arginines. Over a one year period we collected serial serum samples from these unimmunized, specific pathogen-free mice and found that more than one-fifth of them contained elevated levels of dsDNA-binding IgG, but not IgM; whereas mice with a wild type DH sequence did not. Thus, germline bias against the use of arginine enriched DH sequence helps to reduce the likelihood of producing self-reactive antibodies.
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Aaron
authors.initials: A
authors.lastname: Silva-Sanchez
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Cun Ren
authors.initials: CR
authors.lastname: Liu
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America; Program in Immunobiology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Andre M
authors.initials: AM
authors.lastname: Vale
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America; Genetic Engineering Division, National Research Center of Egypt, Ad Doqi, Egypt.
authors.forename: Mohamed
authors.initials: M
authors.lastname: Khass
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Pratibha
authors.initials: P
authors.lastname: Kapoor
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Ada
authors.initials: A
authors.lastname: Elgavish
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Ivaylo I
authors.initials: II
authors.lastname: Ivanov
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Ippolito
authors.affiliation: Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Schelonka
authors.affiliation: Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Trenton R
authors.initials: TR
authors.lastname: Schoeb
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America; Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Peter D
authors.initials: PD
authors.lastname: Burrows
authors.affiliation: Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America; Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, United States of America; Department of Genetics, University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Harry W
authors.initials: HW
authors.lastname: Schroeder
mesh_terms: D000818:Animals; D000916:Antibody Diversity / Q000235:genetics / Q000276:immunology; D001323:Autoantibodies / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D005075:Biological Evolution; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology; D017124:Conserved Sequence / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000235:genetics* / Q000276:immunology*; D005803:Genes, Immunoglobulin / Q000235:genetics* / Q000276:immunology; D007074:Immunoglobulin G / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D016364:Reading Frames / Q000235:genetics / Q000276:immunology; D060152:V(D)J Recombination / Q000235:genetics / Q000276:immunology
chemical_list: D001323:Autoantibodies; D022801:Complementarity Determining Regions; D007074:Immunoglobulin G; D004247:DNA