anti-FcγRIIB (CD32) antibodies differentially modulate murine FVIII-specific recall response in vitro
anti-FcγRIIB (CD32) antibodies differentially modulate murine FVIII-specific recall response in vitro
Fc gamma receptors (FcγRs) for IgG regulate adaptive immune responses by modulating activating and inhibitory signaling pathways within immune cells. Data from a hemophilia A mouse model demonstrate that genetic deletion or blockade of the inhibitory FcγR (CD32) suppresses the formation of antibody secreting cells (ASCs) in vitro. Mechanisms preventing the FVIII-specific recall response, however, remain unclear. Here, the potential role of CD32 inhibition was studied by differentially modulating receptor activity with selected anti-CD32 monoclonal antibodies (mAbs). Splenocytes from immunized FVIII-/- mice were re-stimulated with FVIII in the absence or presence of different anti-CD32 mAbs over 6 days. At day 6, cytokine release was quantified from cell culture supernatant and the formation of FVIII-specific ASCs assessed. Binding of FVIII-containing immune complexes (F8-ICs) to bone marrow derived dendritic cells (BMdDCs) was also investigated. The antagonistic CD32 mAb AT128 suppressed the formation of FVIII-specific ASCs and reduced secretion of IFN-γ and IL-10. In contrast, the agonistic mAbs AT130-2 and AT130-5, and their F(ab')2 fragments, allowed the formation of FVIII-specific ASCs, even though the full IgG of AT130-2 reduced binding of F8-ICs to CD32. Data suggest that an inhibitory signal is transmitted when F8-ICs bind to CD32 and that this signal is required during memory B cell (MBC) activation to support formation of FVIII-specific ASCs. If the inhibitory signal is lacking due to CD32 deletion or blockade with antagonistic anti-CD32 mAbs, FVIII-specific T cell stimulation and ASC formation are suppressed, whereas agonistic stimulation of CD32 restores T cell stimulation and ASC formation.
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Vollack, Nadine
83a4e05c-e998-43c7-895e-c2c3a89aa411
Friese, Julia
df878d6b-bc06-462e-bf79-389a3b5f6b44
Bergmann, Sabine
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Tiede, Andreas
e2eda2ae-c828-43de-813c-6f53d9ce3f3e
Werwitzke, Sonja
cecb3f8b-afc5-44b7-9d3c-bcf20a06e770
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Vollack, Nadine
83a4e05c-e998-43c7-895e-c2c3a89aa411
Friese, Julia
df878d6b-bc06-462e-bf79-389a3b5f6b44
Bergmann, Sabine
2d0dcad5-dc7f-4bac-8018-50709a1ade01
Tiede, Andreas
e2eda2ae-c828-43de-813c-6f53d9ce3f3e
Werwitzke, Sonja
cecb3f8b-afc5-44b7-9d3c-bcf20a06e770
Cragg, Mark, Vollack, Nadine, Friese, Julia, Bergmann, Sabine, Tiede, Andreas and Werwitzke, Sonja
(2017)
anti-FcγRIIB (CD32) antibodies differentially modulate murine FVIII-specific recall response in vitro.
Scandinavian Journal of Immunology.
(doi:10.1111/sji.12573).
Abstract
Fc gamma receptors (FcγRs) for IgG regulate adaptive immune responses by modulating activating and inhibitory signaling pathways within immune cells. Data from a hemophilia A mouse model demonstrate that genetic deletion or blockade of the inhibitory FcγR (CD32) suppresses the formation of antibody secreting cells (ASCs) in vitro. Mechanisms preventing the FVIII-specific recall response, however, remain unclear. Here, the potential role of CD32 inhibition was studied by differentially modulating receptor activity with selected anti-CD32 monoclonal antibodies (mAbs). Splenocytes from immunized FVIII-/- mice were re-stimulated with FVIII in the absence or presence of different anti-CD32 mAbs over 6 days. At day 6, cytokine release was quantified from cell culture supernatant and the formation of FVIII-specific ASCs assessed. Binding of FVIII-containing immune complexes (F8-ICs) to bone marrow derived dendritic cells (BMdDCs) was also investigated. The antagonistic CD32 mAb AT128 suppressed the formation of FVIII-specific ASCs and reduced secretion of IFN-γ and IL-10. In contrast, the agonistic mAbs AT130-2 and AT130-5, and their F(ab')2 fragments, allowed the formation of FVIII-specific ASCs, even though the full IgG of AT130-2 reduced binding of F8-ICs to CD32. Data suggest that an inhibitory signal is transmitted when F8-ICs bind to CD32 and that this signal is required during memory B cell (MBC) activation to support formation of FVIII-specific ASCs. If the inhibitory signal is lacking due to CD32 deletion or blockade with antagonistic anti-CD32 mAbs, FVIII-specific T cell stimulation and ASC formation are suppressed, whereas agonistic stimulation of CD32 restores T cell stimulation and ASC formation.
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Accepted/In Press date: 18 May 2017
e-pub ahead of print date: 13 July 2017
Organisations:
Cancer Sciences
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Local EPrints ID: 411761
URI: http://eprints.soton.ac.uk/id/eprint/411761
ISSN: 0300-9475
PURE UUID: 8aa20c30-14b7-4e34-a5f8-c649c1b03fd4
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Date deposited: 23 Jun 2017 16:31
Last modified: 16 Mar 2024 05:28
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Author:
Nadine Vollack
Author:
Julia Friese
Author:
Sabine Bergmann
Author:
Andreas Tiede
Author:
Sonja Werwitzke
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