Abstract
Objective Using phage display, we sought to screen single-chain variable fragments (scFvs) against complement C5 to treat neuromyelitis optica spectrum disorder (NMOSD).
Methods After 5 rounds of phage display, we isolated individual clones and identified phage clones specifically binding to C5 using ELISA. Using aquaporin-4 (AQP4)-transfected cells in vitro, we confirmed whether these scFvs prevented complement-dependent cytotoxicity (CDC) caused by the serum of patients with NMOSD and human complement (hC). We selected an NMOSD mouse model, in which intracerebral NMOSD immunoglobulin G (IgG) and hC injections induce NMOSD-like lesions in vivo.
Results We obtained scFvs to test specificity and blocking efficiency. The scFv C5B3 neutralized C5 in the complement activation pathway, which prevented AQP4-IgG–mediated CDC in AQP4-transfected cells. In an NMOSD mouse model, C5B3 prevented AQP4 and astrocyte loss, decreased demyelination, and reduced inflammatory infiltration and membrane attack complex formation in lesions.
Conclusions We used phage display to screen C5B3 against C5, which was effective in inhibiting cytotoxicity in vitro and preventing CNS pathology in vivo.
Glossary
- AQP=
- aquaporin-4;
- BSA=
- bovine serum albumin;
- CDC=
- complement-dependent cytotoxicity;
- CHO=
- Chinese hamster ovary;
- GFAP=
- glial fibrillary acidic protein;
- hC=
- human complement;
- HAMA=
- human anti-mouse antibody;
- IBA=
- ionized calcium-binding adaptor molecule;
- IgG=
- immunoglobulin G;
- MAC=
- membrane attack complex;
- MBP=
- myelin basic protein;
- NMOSD=
- neuromyelitis optica spectrum disorder;
- PBS=
- phosphate-buffered saline;
- PFA=
- paraformaldehyde;
- SPR=
- surface plasmon resonance
Footnotes
Funding information and disclosures are provided at the end of the article. Full disclosure form information provided by the authors is available with the full text of this article at Neurology.org/NN.
The Article Processing Charge was funded by the authors.
- Received September 17, 2018.
- Accepted in final form February 5, 2019.
- Copyright © 2019 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Academy of Neurology.
This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND), which permits downloading and sharing the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.
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