Ex vivo assays to detect complement activation in complementopathies.

ABSTRACT: In complement-driven thrombotic microangiopathies, failure to regulate complement activation leads to end-organ damage. The modified Ham (mHam) test measures complement-mediated killing of a nucleated cell in vitro but lacks a confirmatory assay and reliable positive controls. We demonstrate that C5b-9 accumulation on the surface of TF1 PIGAnull cells correlates with cell killing in the mHam. We also show that Sialidase treatment of cells or addition of Shiga toxin 1 to human serum serve as a more reliable positive control for the mHam than cobra venom factor or lipopolysaccharide. Simultaneously performing the mHam and measuring C5b-9 accumulation either in GVB⁺⁺ or GVB⁰ MgEGTA buffer with the addition of complement pathway specific inhibitors (anti-C5 antibody or a

SUBMITTER: Yuan X 

PROVIDER: S-EPMC8609776 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature