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Shaving is an epiphenomenon of Type i and II anti-CD20-mediated phagocytosis, whereas antigenic modulation limits type i monoclonal antibody efficacy

Shaving is an epiphenomenon of Type i and II anti-CD20-mediated phagocytosis, whereas antigenic modulation limits type i monoclonal antibody efficacy
Shaving is an epiphenomenon of Type i and II anti-CD20-mediated phagocytosis, whereas antigenic modulation limits type i monoclonal antibody efficacy

Rituximab is an anti-CD20 mAb used in the treatment of B cell malignancies. Loss of surface CD20 Ag from the surface of target cells is thought to be one mechanism governing resistance to rituximab, but how this occurs is not completely understood. Two explanations for this have been proposed: antigenic modulation whereby mAb:CD20 complexes are internalized in a B cell intrinsic process and shaving, in which mAb:CD20 complexes undergo trogocytic removal by effector cells, such as macrophages. However, there is conflicting evidence as to which predominates in clinical scenarios and hence the best strategies to overcome resistance. In this study, we investigated the relative importance of modulation and shaving in the downregulation of surface mAb:CD20. We used both murine and human systems and treated ex vivo macrophages with varying concentrations of non-FcgR-interacting beads to achieve differential macrophage saturation states, hence controllably suppressing further phagocytosis of target cells. We then monitored the level and localization of mAb:CD20 using a quenching assay. Suppression of phagocytosis with bead treatment decreased shaving and increased modulation, suggesting that the two compete for surface rituximab:CD20. Under all conditions tested, modulation predominated in rituximab loss, whereas shaving represented an epiphenomenon to phagocytosis. We also demonstrate that the nonmodulating, glycoengineered, type II mAb obinutuzumab caused a modest but significant increase in shaving compared with type II BHH2 human IgG1 wild-type mAb. Therefore, shaving may represent an important mechanism of resistance when modulation is curtailed, and glycoengineering mAb to increase affinity for FcgR may enhance resistance because of shaving.

0022-1767
1211-1221
Dahal, Lekh N.
1e993a7a-b007-4187-82ea-e28dd3920b66
Huang, Chie Yin
6964b72b-71cd-4499-8a01-7ebd4840fae2
Stopforth, Richard J.
2d3e18ff-5563-4247-9150-f0f337fb585f
Mead, Abbie
7246f2f0-c9ac-40f8-8b31-ba0e73beaff0
Chan, Keith
87a1f006-bba1-4ca9-bd19-1dde044f714e
Bowater, Juliet X.
a6226c0b-e187-460b-943c-fad7fd4162e7
Taylor, Martin C.
9e6d2e6d-5e00-4610-8ec6-5f117296124a
Narang, Priyanka
b2e8fcbd-c75a-444b-b95f-39dea0bc5d3d
Chan, H. T.Claude
9b7d97e7-f135-478b-883e-3ff52d49af85
Kim, Jinny H.
ea81033c-8f9b-427b-be10-7a320a3f2651
Vaughan, Andrew T.
bfb2ceab-a592-457e-89f9-00fcd1dddbdb
Forconi, Francesco
ce9ed873-58cf-4876-bf3a-9ba1d163edc8
Beers, Stephen A.
a02548be-3ffd-41ab-9db8-d6e8c3b499a2
Dahal, Lekh N.
1e993a7a-b007-4187-82ea-e28dd3920b66
Huang, Chie Yin
6964b72b-71cd-4499-8a01-7ebd4840fae2
Stopforth, Richard J.
2d3e18ff-5563-4247-9150-f0f337fb585f
Mead, Abbie
7246f2f0-c9ac-40f8-8b31-ba0e73beaff0
Chan, Keith
87a1f006-bba1-4ca9-bd19-1dde044f714e
Bowater, Juliet X.
a6226c0b-e187-460b-943c-fad7fd4162e7
Taylor, Martin C.
9e6d2e6d-5e00-4610-8ec6-5f117296124a
Narang, Priyanka
b2e8fcbd-c75a-444b-b95f-39dea0bc5d3d
Chan, H. T.Claude
9b7d97e7-f135-478b-883e-3ff52d49af85
Kim, Jinny H.
ea81033c-8f9b-427b-be10-7a320a3f2651
Vaughan, Andrew T.
bfb2ceab-a592-457e-89f9-00fcd1dddbdb
Forconi, Francesco
ce9ed873-58cf-4876-bf3a-9ba1d163edc8
Beers, Stephen A.
a02548be-3ffd-41ab-9db8-d6e8c3b499a2

Dahal, Lekh N., Huang, Chie Yin, Stopforth, Richard J., Mead, Abbie, Chan, Keith, Bowater, Juliet X., Taylor, Martin C., Narang, Priyanka, Chan, H. T.Claude, Kim, Jinny H., Vaughan, Andrew T., Forconi, Francesco and Beers, Stephen A. (2018) Shaving is an epiphenomenon of Type i and II anti-CD20-mediated phagocytosis, whereas antigenic modulation limits type i monoclonal antibody efficacy. The Journal of Immunology, 201 (4), 1211-1221. (doi:10.4049/jimmunol.1701122).

Record type: Article

Abstract

Rituximab is an anti-CD20 mAb used in the treatment of B cell malignancies. Loss of surface CD20 Ag from the surface of target cells is thought to be one mechanism governing resistance to rituximab, but how this occurs is not completely understood. Two explanations for this have been proposed: antigenic modulation whereby mAb:CD20 complexes are internalized in a B cell intrinsic process and shaving, in which mAb:CD20 complexes undergo trogocytic removal by effector cells, such as macrophages. However, there is conflicting evidence as to which predominates in clinical scenarios and hence the best strategies to overcome resistance. In this study, we investigated the relative importance of modulation and shaving in the downregulation of surface mAb:CD20. We used both murine and human systems and treated ex vivo macrophages with varying concentrations of non-FcgR-interacting beads to achieve differential macrophage saturation states, hence controllably suppressing further phagocytosis of target cells. We then monitored the level and localization of mAb:CD20 using a quenching assay. Suppression of phagocytosis with bead treatment decreased shaving and increased modulation, suggesting that the two compete for surface rituximab:CD20. Under all conditions tested, modulation predominated in rituximab loss, whereas shaving represented an epiphenomenon to phagocytosis. We also demonstrate that the nonmodulating, glycoengineered, type II mAb obinutuzumab caused a modest but significant increase in shaving compared with type II BHH2 human IgG1 wild-type mAb. Therefore, shaving may represent an important mechanism of resistance when modulation is curtailed, and glycoengineering mAb to increase affinity for FcgR may enhance resistance because of shaving.

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JI_Manuscript_June_2018 - Accepted Manuscript
Restricted to Repository staff only until 10 June 2019.
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More information

Accepted/In Press date: 10 June 2018
e-pub ahead of print date: 6 August 2018
Published date: 15 August 2018

Identifiers

Local EPrints ID: 423432
URI: https://eprints.soton.ac.uk/id/eprint/423432
ISSN: 0022-1767
PURE UUID: 9a57c8e7-f1c4-4e88-a856-87ca24dc591f
ORCID for Abbie Mead: ORCID iD orcid.org/0000-0002-1299-8512
ORCID for Andrew T. Vaughan: ORCID iD orcid.org/0000-0001-6076-3649

Catalogue record

Date deposited: 21 Sep 2018 16:30
Last modified: 14 Mar 2019 01:38

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