Biochemical analysis of proteasomes from mouse microglia: induction of immunoproteasomes by interferon-gamma and lipopolysaccharide

Glia. 2000 Feb 15;29(4):355-65.

Abstract

The 20S proteasome is a multicatalytic threonine protease and serves to process peptides that are subsequently presented as antigenic epitopes by MHC class I molecules. In the brain, microglial cells are the major antigen presenting cells and they respond sensitive to pathologic events. We used cultured mouse microglia and a microglial cell line, the BV-2 line, as a model to study the correlation between microglial activation parameters and structural plasticity of the 20S/26S proteasome. Lipopolysaccharide (LPS)- or interferon-gamma (IFN-gamma)-stimulated microglia or BV-2 cells exhibit properties of activated microglia such as high levels of TNFalpha and IL-6 release. In response to IFN-gamma or LPS, three constitutive beta subunits (beta1/Delta, beta2/MC14, beta5/MB1) were replaced by the immunoproteasome subunits ibeta1/LMP2, ibeta2/MECL-1, and ibeta5/LMP7, indicating that activated microglia adapts its proteasomal subunit composition to the requirements of an optimized MHC class I epitope processing. Induction of immunoproteasomes in BV-2 cells was solely provoked by IFN-gamma, but not by LPS. Moreover, LPS (but not IFN-gamma) triggered the expression of a novel protein of approximately 50 kD as part of the proteasome activator PA700, that is the substrate-recognizing and unfolding unit of the 26S proteasome. These results indicate that both the 20S core protease as well as the proteasome activator PA700 are targets of modulatory subunit replacements or transient association of regulatory components in the course of microglial activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Animals
  • Antigen Presentation
  • Antioxidants / pharmacology
  • Ascorbic Acid / pharmacology
  • Cells, Cultured
  • Cysteine Endopeptidases / chemistry*
  • Cysteine Endopeptidases / drug effects
  • Cysteine Endopeptidases / ultrastructure
  • Electrophoresis, Gel, Two-Dimensional
  • Interferon-gamma / pharmacology*
  • Interleukin-6 / metabolism
  • Lipopolysaccharides / pharmacology*
  • Mice
  • Mice, Inbred Strains
  • Microglia / immunology*
  • Microglia / metabolism
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / drug effects
  • Multienzyme Complexes / ultrastructure
  • Phenotype
  • Precipitin Tests
  • Proteasome Endopeptidase Complex
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Antioxidants
  • Interleukin-6
  • Lipopolysaccharides
  • Multienzyme Complexes
  • Tumor Necrosis Factor-alpha
  • Interferon-gamma
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • Adenosine Triphosphatases
  • Ascorbic Acid