Ex vivo assays to detect complement activation in complementopathies.

Xuan Yuan, Jia Yu, Gloria Gerber, Shruti Chaturvedi, Michael Cole, Hang Chen, Ara Metjian, C John Sperati, Evan M Braunstein, Robert A Brodsky

Clinical immunology (Orlando, Fla.) 2020 Dec

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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. Copyright © 2020. Published by Elsevier Inc.

Citation

Xuan Yuan, Jia Yu, Gloria Gerber, Shruti Chaturvedi, Michael Cole, Hang Chen, Ara Metjian, C John Sperati, Evan M Braunstein, Robert A Brodsky. Ex vivo assays to detect complement activation in complementopathies. Clinical immunology (Orlando, Fla.). 2020 Dec;221:108616