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Area of Science:

  • Immunology
  • Molecular Biology

Background:

  • Bacterial and mitochondrial DNA, containing CpG motifs, are danger signals triggering inflammation.
  • Mechanisms of complement activation by CpG motifs are not fully understood.

Purpose of the Study:

  • To investigate how synthetic CpG oligodeoxynucleotides activate the complement system.
  • To identify the specific components and pathways involved in CpG-mediated complement activation.

Main Methods:

  • Incubation of human whole blood and plasma with synthetic CpG-A, -B, -C, and GpC controls.
  • Analysis of complement activation products (C5b-9, C3bBbP, C3bc) and cytokine levels (27-plex immunoassay).
  • Evaluation of CpG-B binding to complement proteins (C1q, Factor H) and complement activation in deficient plasma/serum.

Main Results:

  • CpG-B and CpG-C, but not CpG-A, dose-dependently activated complement.
  • Complement activation was dependent on the DNA backbone, not CpG motifs.
  • CpG-B bound to C1q and Factor H, and activation was partly preserved in C2-deficient plasma and MASP-2-deficient serum.
  • CpG-B induced cytokine release (IL-1β, IL-6, TNF, etc.) dependent on C5aR1.

Conclusions:

  • Synthetic CpG DNA activates the complement system via its DNA backbone.
  • CpG-B interacts with complement components, leading to C5aR1-dependent cytokine release, linking innate immune sensing to inflammation.