Ex Vivo Assays to Detect Complement Activation in Complementopathies

Overview

Journal Clin Immunol

Specialty Allergy & Immunology

Date 2020 Nov 5

PMID 33148511

Citations 7

Authors

Xuan Yuan

Jia Yu

Gloria Gerber

Shruti Chaturvedi

Michael Cole

Hang Chen

Ara Metjian

C John Sperati

Evan M Braunstein

Robert A Brodsky

Affiliations

Soon will be listed here.

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 GVB 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.

Citing Articles

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Can complement activation be the missing link in antiphospholipid syndrome?.

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Novel Functional Assay to Characterize Mutations in Alternative Pathway of Complement.

Gerber G, Pan X, Lederman H, Brady T, Brodsky R J Clin Immunol. 2023; 43(6):1134-1136.

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