A new panel of epitope mapped monoclonal antibodies recognising the prototypical tetraspanin CD81

Grove, J. , Hu, K., Farquhar, M. J., Goodall, M., Walker, L., Jamshad, M., Drummer, H. E., Bill, R. M., Balfe, P. and McKeating, J. A. (2017) A new panel of epitope mapped monoclonal antibodies recognising the prototypical tetraspanin CD81. Wellcome Open Research, 2, 82. (doi: 10.12688/wellcomeopenres.12058.1) (PMID:29090272) (PMCID:PMC5657224)

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Abstract

Background: Tetraspanins are small transmembrane proteins, found in all higher eukaryotes, that compartmentalize cellular membranes through interactions with partner proteins. CD81 is a prototypical tetraspanin and contributes to numerous physiological and pathological processes, including acting as a critical entry receptor for hepatitis C virus (HCV). Antibody engagement of tetraspanins can induce a variety of effects, including actin cytoskeletal rearrangements, activation of MAPK-ERK signaling and cell migration. However, the epitope specificity of most anti-tetraspanin antibodies is not known, limiting mechanistic interpretation of these studies. Methods: We generated a panel of monoclonal antibodies (mAbs) specific for CD81 second extracellular domain (EC2) and performed detailed epitope mapping with a panel of CD81 mutants. All mAbs were screened for their ability to inhibit HCV infection and E2-CD81 association. Nanoscale distribution of cell surface CD81 was investigated by scanning electron microscopy. Results: The antibodies were classified in two epitope groups targeting opposing sides of EC2. We observed a wide range of anti-HCV potencies that were independent of their epitope grouping, but associated with their relative affinity for cell-surface expressed CD81. Scanning electron microscopy identified at least two populations of CD81; monodisperse and higher-order assemblies, consistent with tetraspanin-enriched microdomains. Conclusions: These novel antibodies provide well-characterised tools to investigate CD81 function, including HCV entry, and have the potential to provide insights into tetraspanin biology in general.

Item Type:Articles
Additional Information:Version 1; peer review: 1 approved, 2 approved with reservations. This work was supported by the Biotechnology and Biological Sciences Research Council (grant number BB/N007417/1, RMB and JAM); the European Commission (via contract LSHG-CT-2004-504601; E-MeP, RMB), the Medical Research Council core funding to the MRC-UCL Laboratory for Molecular Cell Biology University Unit (award code MC_UU_12018/1, JG); The Grove Lab is supported by the Wellcome Trust and Royal Society (grant number 107653). Research in the McKeating laboratory is funded by MRC Programme G1100247, EU FP7 funded PathCO HEALTH F3-2012-305578 and the Wellcome Trust (grant number 200838).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Grove, Dr Joe
Creator Roles:
Grove, J.Investigation, Writing – original draft
Authors: Grove, J., Hu, K., Farquhar, M. J., Goodall, M., Walker, L., Jamshad, M., Drummer, H. E., Bill, R. M., Balfe, P., and McKeating, J. A.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
Journal Name:Wellcome Open Research
Publisher:F1000Research
ISSN:2398-502X
ISSN (Online):2398-502X
Copyright Holders:Copyright © 2017 Grove J et al.
First Published:First published in Wellcome Open Research 2: 82
Publisher Policy:Reproduced under a Creative Commons License

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