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 factor D inhibitor, ACH-145951) can be used to localize defects in complement regulation. As more targeted complement inhibitors become available, these assays may aid in the selection of personalized treatments for patients with complement-mediated diseases.
Original language | English (US) |
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Article number | 108616 |
Journal | Clinical Immunology |
Volume | 221 |
DOIs | |
State | Published - Dec 2020 |
Keywords
- Atypical hemolytic uremic syndrome
- Complement
- Complement inhibitors
- Modified HAM assay
- Shiga toxin 1
- Sialidase
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology