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Development and characterisation of monoclonal antibodies specific for the murine inhibitory Fc?RIIB (CD32B)

Development and characterisation of monoclonal antibodies specific for the murine inhibitory Fc?RIIB (CD32B)
Development and characterisation of monoclonal antibodies specific for the murine inhibitory Fc?RIIB (CD32B)
Fc receptors (FcRs) play a key role in regulating and coordinating responses from both innate and adaptive arms of the immune system. The inhibitory Fc gamma receptor II (Fc?RIIB; CD32) is central to this regulation with Fc?RIIB(-/-) mice demonstrating augmented responses to mAb immunotherapy, elevated incidence and severity of auto-immunity, and increased response to mAb-mediated cancer therapy. To date, these observations have remained unexploited therapeutically, partly through a lack of specific mAb reagents capable of exclusively binding mouse Fc?RIIB. Thus almost all of the Fc?RIIB-binding mAb currently available, such as 2.4G2, also bind Fc?RIII (CD16), and polyclonal reagents have limited availability and are of unproven specificity and avidity, making in vivo manipulation of Fc?RIIB impossible. Following an extensive immunisation protocol using Fc?RIIB(-/-) mice, we recently produced three unique mAb that are suitable for this purpose. Here we characterise these novel reagents and demonstrate that they fall into two distinct categories; those which cause phosphorylation and subsequent activation of Fc?RIIB (agonistic) and those that block receptor phosphorylation (antagonistic). These two types of mAb exhibit different characteristics in a range of biochemical, cellular, and functional assays relevant to Fc?RIIB activity and mAb therapy.
antibodies, cancer, CD32B, Fc gamma RIIB, immunotherapy
0014-2980
2109-20
Williams, Emily L.
f2650481-4f32-470a-b7fb-c78f026de16c
Tutt, Alison L.
956cf1ab-867c-4db7-b0d1-93d408781fb8
French, Ruth R.
cf2ef484-f0f9-47d0-8461-5fb71d950be3
Chan, H.T. Claude
b109c93f-7e9a-44ee-ad12-da757b1b11fc
Lau, Betty
40c845a2-a710-4635-a596-dd1fe6eafa11
Penfold, Christine A.
273343c6-045f-4bd7-9878-7e9513344d65
Mockridge, C. Ian
327aef17-4837-4f2a-a93b-3d17cd1a7f9f
Roghanian, Ali
e2b032c2-60a0-4522-a3d8-56a768792f36
Cox, Kerry L.
7305c27e-9cdc-4e37-b994-ac55d7d1dfd2
Verbeek, J. Sjef
115ffb7c-4760-444f-888c-0798469e0b9c
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded
Cragg, Mark S.
ec97f80e-f3c8-49b7-a960-20dff648b78c
Williams, Emily L.
f2650481-4f32-470a-b7fb-c78f026de16c
Tutt, Alison L.
956cf1ab-867c-4db7-b0d1-93d408781fb8
French, Ruth R.
cf2ef484-f0f9-47d0-8461-5fb71d950be3
Chan, H.T. Claude
b109c93f-7e9a-44ee-ad12-da757b1b11fc
Lau, Betty
40c845a2-a710-4635-a596-dd1fe6eafa11
Penfold, Christine A.
273343c6-045f-4bd7-9878-7e9513344d65
Mockridge, C. Ian
327aef17-4837-4f2a-a93b-3d17cd1a7f9f
Roghanian, Ali
e2b032c2-60a0-4522-a3d8-56a768792f36
Cox, Kerry L.
7305c27e-9cdc-4e37-b994-ac55d7d1dfd2
Verbeek, J. Sjef
115ffb7c-4760-444f-888c-0798469e0b9c
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded
Cragg, Mark S.
ec97f80e-f3c8-49b7-a960-20dff648b78c

Williams, Emily L., Tutt, Alison L., French, Ruth R., Chan, H.T. Claude, Lau, Betty, Penfold, Christine A., Mockridge, C. Ian, Roghanian, Ali, Cox, Kerry L., Verbeek, J. Sjef, Glennie, Martin J. and Cragg, Mark S. (2012) Development and characterisation of monoclonal antibodies specific for the murine inhibitory Fc?RIIB (CD32B). European Journal of Immunology, 42 (8), 2109-20. (doi:10.1002/eji.201142302). (PMID:22760702)

Record type: Article

Abstract

Fc receptors (FcRs) play a key role in regulating and coordinating responses from both innate and adaptive arms of the immune system. The inhibitory Fc gamma receptor II (Fc?RIIB; CD32) is central to this regulation with Fc?RIIB(-/-) mice demonstrating augmented responses to mAb immunotherapy, elevated incidence and severity of auto-immunity, and increased response to mAb-mediated cancer therapy. To date, these observations have remained unexploited therapeutically, partly through a lack of specific mAb reagents capable of exclusively binding mouse Fc?RIIB. Thus almost all of the Fc?RIIB-binding mAb currently available, such as 2.4G2, also bind Fc?RIII (CD16), and polyclonal reagents have limited availability and are of unproven specificity and avidity, making in vivo manipulation of Fc?RIIB impossible. Following an extensive immunisation protocol using Fc?RIIB(-/-) mice, we recently produced three unique mAb that are suitable for this purpose. Here we characterise these novel reagents and demonstrate that they fall into two distinct categories; those which cause phosphorylation and subsequent activation of Fc?RIIB (agonistic) and those that block receptor phosphorylation (antagonistic). These two types of mAb exhibit different characteristics in a range of biochemical, cellular, and functional assays relevant to Fc?RIIB activity and mAb therapy.

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More information

e-pub ahead of print date: 4 July 2012
Published date: August 2012
Keywords: antibodies, cancer, CD32B, Fc gamma RIIB, immunotherapy
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 369615
URI: http://eprints.soton.ac.uk/id/eprint/369615
ISSN: 0014-2980
PURE UUID: 0f274629-3570-4e34-b604-330bf50080ed
ORCID for H.T. Claude Chan: ORCID iD orcid.org/0000-0003-0530-9480
ORCID for Ali Roghanian: ORCID iD orcid.org/0000-0003-1316-4218
ORCID for Mark S. Cragg: ORCID iD orcid.org/0000-0003-2077-089X

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Date deposited: 02 Oct 2014 09:50
Last modified: 15 Mar 2024 03:34

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Contributors

Author: Emily L. Williams
Author: Alison L. Tutt
Author: Ruth R. French
Author: H.T. Claude Chan ORCID iD
Author: Betty Lau
Author: Christine A. Penfold
Author: C. Ian Mockridge
Author: Ali Roghanian ORCID iD
Author: Kerry L. Cox
Author: J. Sjef Verbeek
Author: Mark S. Cragg ORCID iD

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