title: Heavy-light chain interrelations of MS-associated immunoglobulins probed by deep sequencing and rational variation.
abstract: The mechanisms triggering most of autoimmune diseases are still obscure. Autoreactive B cells play a crucial role in the development of such pathologies and, in particular, production of autoantibodies of different specificities. The combination of deep-sequencing technology with functional studies of antibodies selected from highly representative immunoglobulin combinatorial libraries may provide unique information on specific features in the repertoires of autoreactive B cells. Here, we have analyzed cross-combinations of the variable regions of human immunoglobulins against the myelin basic protein (MBP) previously selected from a multiple sclerosis (MS)-related scFv phage-display library. On the other hand, we have performed deep sequencing of the sublibraries of scFvs against MBP, Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1), and myelin oligodendrocyte glycoprotein (MOG). Bioinformatics analysis of sequencing data and surface plasmon resonance (SPR) studies have shown that it is the variable fragments of antibody heavy chains that mainly determine both the affinity of antibodies to the parent autoantigen and their cross-reactivity. It is suggested that LMP1-cross-reactive anti-myelin autoantibodies contain heavy chains encoded by certain germline gene segments, which may be a hallmark of the EBV-specific B cell subpopulation involved in MS triggering.
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Yakov A
authors.initials: YA
authors.lastname: Lomakin
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Maria Yu
authors.initials: MY
authors.lastname: Zakharova
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Alexey V
authors.initials: AV
authors.lastname: Stepanov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Dronina
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
authors.forename: Ivan V
authors.initials: IV
authors.lastname: Smirnov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Tatyana V
authors.initials: TV
authors.lastname: Bobik
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Andrey Yu
authors.initials: AY
authors.lastname: Pyrkov
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: Nina V
authors.initials: NV
authors.lastname: Tikunova
authors.affiliation: Russian State Medical University, Department of Neurology & Neurosurgery, Moscow Multiple Sclerosis Center at the City Hospital #11, Moscow, Russia.
authors.forename: Svetlana N
authors.initials: SN
authors.lastname: Sharanova
authors.affiliation: St. Petersburg Academic University, Nanotechnology Research and Education Centre, Russian Academy of Sciences, St. Petersburg, Russia.
authors.forename: Vitali M
authors.initials: VM
authors.lastname: Boitsov
authors.affiliation: St. Petersburg Academic University, Nanotechnology Research and Education Centre, Russian Academy of Sciences, St. Petersburg, Russia.
authors.forename: Sergey Yu
authors.initials: SY
authors.lastname: Vyazmin
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia; Genomics Core Facility, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: Marsel R
authors.initials: MR
authors.lastname: Kabilov
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia; Genomics Core Facility, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia; Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Alexey E
authors.initials: AE
authors.lastname: Tupikin
authors.affiliation: Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Alexey N
authors.initials: AN
authors.lastname: Krasnov
authors.affiliation: Kharkevich Institute for Information Transmission Problems, Russian Academy of Sciences, Moscow, Russia; Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia.
authors.forename: Nadezda A
authors.initials: NA
authors.lastname: Bykova
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; Institut de Medicina Predictiva I Personalitzada del Càncer, Badalona (Barcelona), Spain.
authors.forename: Yulia A
authors.initials: YA
authors.lastname: Medvedeva
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Fridman
authors.affiliation: Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia; Department of Oncology, Division of Biostatistics and Bioinformatics, Johns Hopkins University School of Medicine, Baltimore, MD, USA; State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIGenetika, Moscow, Russia.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Favorov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Ponomarenko
authors.affiliation: St. Petersburg Academic University, Nanotechnology Research and Education Centre, Russian Academy of Sciences, St. Petersburg, Russia.
authors.forename: Michael V
authors.initials: MV
authors.lastname: Dubina
authors.affiliation: Russian State Medical University, Department of Neurology & Neurosurgery, Moscow Multiple Sclerosis Center at the City Hospital #11, Moscow, Russia.
authors.forename: Alexey N
authors.initials: AN
authors.lastname: Boyko
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia; Novosibirsk State University, Novosibirsk, Russia.
authors.forename: Valentin V
authors.initials: VV
authors.lastname: Vlassov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Belogurov
authors.affiliation: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia; Faculty of Chemistry, Lomonosov Moscow State University, Moscow, Russia; Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia. Electronic address: gabibov@mx.ibch.ru.
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Gabibov
mesh_terms: D001323:Autoantibodies / Q000276:immunology; D001327:Autoimmune Diseases / Q000276:immunology; D003429:Cross Reactions; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D007136:Immunoglobulins / Q000276:immunology*; D009103:Multiple Sclerosis / Q000276:immunology*; D004676:Myelin Basic Protein / Q000276:immunology; D063308:Myelin-Oligodendrocyte Glycoprotein / Q000276:immunology; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D001323:Autoantibodies; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D007143:Immunoglobulin Heavy Chains; D007136:Immunoglobulins; C499048:MBP protein, human; C571326:MOG protein, human; D004676:Myelin Basic Protein; D063308:Myelin-Oligodendrocyte Glycoprotein; D014763:Viral Matrix Proteins
keywords: Antibody cross-reactivity; B cells deep sequencing; Epstein–Barr virus; Human germline autoantibody gene segments; Latent membrane protein 1; Multiple sclerosis; Myelin basic protein