Dysregulation of B Cell Repertoire Formation in Myasthenia Gravis Patients Revealed through Deep Sequencing

J Immunol. 2017 Feb 15;198(4):1460-1473. doi: 10.4049/jimmunol.1601415. Epub 2017 Jan 13.

Abstract

Myasthenia gravis (MG) is a prototypical B cell-mediated autoimmune disease affecting 20-50 people per 100,000. The majority of patients fall into two clinically distinguishable types based on whether they produce autoantibodies targeting the acetylcholine receptor (AChR-MG) or muscle specific kinase (MuSK-MG). The autoantibodies are pathogenic, but whether their generation is associated with broader defects in the B cell repertoire is unknown. To address this question, we performed deep sequencing of the BCR repertoire of AChR-MG, MuSK-MG, and healthy subjects to generate ∼518,000 unique VH and VL sequences from sorted naive and memory B cell populations. AChR-MG and MuSK-MG subjects displayed distinct gene segment usage biases in both VH and VL sequences within the naive and memory compartments. The memory compartment of AChR-MG was further characterized by reduced positive selection of somatic mutations in the VH CDR and altered VH CDR3 physicochemical properties. The VL repertoire of MuSK-MG was specifically characterized by reduced V-J segment distance in recombined sequences, suggesting diminished VL receptor editing during B cell development. Our results identify large-scale abnormalities in both the naive and memory B cell repertoires. Particular abnormalities were unique to either AChR-MG or MuSK-MG, indicating that the repertoires reflect the distinct properties of the subtypes. These repertoire abnormalities are consistent with previously observed defects in B cell tolerance checkpoints in MG, thereby offering additional insight regarding the impact of tolerance defects on peripheral autoimmune repertoires. These collective findings point toward a deformed B cell repertoire as a fundamental component of MG.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Autoantibodies / immunology
  • B-Lymphocytes / immunology*
  • B-Lymphocytes / pathology
  • B-Lymphocytes / physiology
  • Female
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Immune Tolerance
  • Immunologic Memory
  • Male
  • Middle Aged
  • Myasthenia Gravis / immunology*
  • Myasthenia Gravis / physiopathology
  • Protein Kinases / immunology
  • Receptor Protein-Tyrosine Kinases / immunology
  • Receptors, Antigen, B-Cell / genetics*
  • Receptors, Antigen, B-Cell / immunology
  • Receptors, Cholinergic / immunology
  • Young Adult

Substances

  • Autoantibodies
  • Receptors, Antigen, B-Cell
  • Receptors, Cholinergic
  • Protein Kinases
  • acetylcholine receptor kinase
  • MUSK protein, human
  • Receptor Protein-Tyrosine Kinases