Broad Susceptibility of Nucleolar Proteins and Autoantigens to Complement C1 Protease Degradation

J Immunol. 2017 Dec 15;199(12):3981-3990. doi: 10.4049/jimmunol.1700728. Epub 2017 Oct 25.

Abstract

Anti-nuclear autoantibodies, which frequently target the nucleoli, are pathogenic hallmarks of systemic lupus erythematosus (SLE). Although the causes of these Abs remain broad and ill-defined, a genetic deficiency in C1 complex (C1qC1r2C1s2) or C4 is able to induce these Abs. Considering a recent finding that, in dead cells, nucleoli were targeted by C1q and two nucleolar autoantigens were degraded by C1r/C1s proteases, we considered that C1 could help protect against antinuclear autoimmunity by broadly degrading nucleolar proteins or autoantigens. Nucleoli were isolated to homogeneity and structurally defined. After C1 treatment, cleaved nucleolar proteins were identified by proteomic two-dimensional fluorescence difference gel electrophoresis and mass spectrometry, and further verified by Western blotting using specific Abs. The extent of nucleolar autoantigen degradation upon C1 treatment was estimated using SLE patient autoantibodies. The isolated nucleoli were broadly reactive with SLE patient autoantibodies. These nucleoli lacked significant autoproteolysis, but many nucleolar proteins and autoantigens were degraded by C1 proteases; >20 nucleolar proteins were identified as C1 cleavable. These were further validated by Western blotting using specific Abs. The broad autoantigenicity of the nucleoli may attribute to their poor autoproteolysis, causing autologous immune stimulation upon necrotic exposure. However, C1q targets at these nucleoli to cause C1 protease activation and the cleavage of many nucleolar proteins or autoantigens. This may represent one important surveillance mechanism against antinuclear autoimmunity because C1 genetic deficiency causes anti-nuclear autoantibodies and SLE disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoantibodies / blood
  • Autoantibodies / immunology*
  • Autoantigens / metabolism*
  • Cell Nucleolus / immunology*
  • Cell Nucleolus / ultrastructure
  • Complement C1r / metabolism*
  • Complement C1s / metabolism*
  • HeLa Cells
  • Humans
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology*
  • Lupus Erythematosus, Systemic / blood
  • Lupus Erythematosus, Systemic / immunology
  • Nuclear Proteins / metabolism*
  • Proteolysis
  • Proteomics
  • Substrate Specificity

Substances

  • Autoantibodies
  • Autoantigens
  • Immunoglobulin G
  • Nuclear Proteins
  • Complement C1r
  • Complement C1s