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Antibody-peptide-MHC fusion conjugates target non-cognate T cells to kill tumour cells

Antibody-peptide-MHC fusion conjugates target non-cognate T cells to kill tumour cells
Antibody-peptide-MHC fusion conjugates target non-cognate T cells to kill tumour cells
Attempts to generate robust anti-tumour cytotoxic T lymphocyte (CTL) responses using immunotherapy are frequently thwarted by exhaustion and anergy of CTL recruited to tumour. One strategy to overcome this is to retarget a population of virus-specific CTL to kill tumour cells. Here, we describe a proof-of-principle study using a bispecific conjugate designed to retarget ovalbumin (OVA)-specific CTL to kill tumour cells via CD20. A single-chain trimer (SCT) consisting of MHCI H-2K(b)/SIINFEKL peptide/beta 2 microglobulin/BirA was expressed in bacteria, refolded and chemically conjugated to one (1:1; F2) or two (2:1; F3) anti-hCD20 Fab' fragments. In vitro, the [SCT × Fab'] (F2 and F3) redirected SIINFEKL-specific OT-I CTL to kill CD20(+) target cells, and in the presence of CD20(+) target cells to provide crosslinking, they were also able to induce proliferation of OT-I cells. In vivo, activated OT-I CTL could be retargeted to kill [SCT × Fab']-coated B cells from hCD20 transgenic (hCD20 Tg) mice and also EL4 and B16 mouse tumour cells expressing human CD20 (hCD20). Importantly, in a hCD20 Tg mouse model, [SCT × Fab'] administered systemically were able to retarget activated OT-I cells to deplete normal B cells, and their performance matched that of a bispecific antibody (BsAb) comprising anti-CD3 and anti-CD20. [SCT × Fab'] were also active therapeutically in an EL4 tumour model. Furthermore, measurement of serum cytokine levels suggests that [SCT × Fab'] are associated with a lower level of inflammatory cytokine release than the BsAb and so may be advantageous clinically in terms of reduced toxicity.
0340-7004
1093-1105
King, Ben C.
aee5fe5f-fc9c-4cc1-8a11-08f099d20a1d
Hamblin, Angela D.
1c6da054-f1e8-4e58-a180-1c99b4f0a33a
Savage, Philip M.
eea178be-7195-4a66-abd5-24b9f4b06bb0
Douglas, Leon R.
049b5f33-6870-4773-ae34-663489b472ba
Hansen, Ted H.
dab738fa-2068-49fa-bf6b-c095e6d6a512
French, Ruth R.
a95ea7a1-7aeb-4c20-998e-fde663613fd1
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded
King, Ben C.
aee5fe5f-fc9c-4cc1-8a11-08f099d20a1d
Hamblin, Angela D.
1c6da054-f1e8-4e58-a180-1c99b4f0a33a
Savage, Philip M.
eea178be-7195-4a66-abd5-24b9f4b06bb0
Douglas, Leon R.
049b5f33-6870-4773-ae34-663489b472ba
Hansen, Ted H.
dab738fa-2068-49fa-bf6b-c095e6d6a512
French, Ruth R.
a95ea7a1-7aeb-4c20-998e-fde663613fd1
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded

King, Ben C., Hamblin, Angela D., Savage, Philip M., Douglas, Leon R., Hansen, Ted H., French, Ruth R., Johnson, Peter W.M. and Glennie, Martin J. (2013) Antibody-peptide-MHC fusion conjugates target non-cognate T cells to kill tumour cells. Cancer Immunology Immunotherapy, 62 (6), 1093-1105. (doi:10.1007/s00262-013-1408-8). (PMID:23604105)

Record type: Article

Abstract

Attempts to generate robust anti-tumour cytotoxic T lymphocyte (CTL) responses using immunotherapy are frequently thwarted by exhaustion and anergy of CTL recruited to tumour. One strategy to overcome this is to retarget a population of virus-specific CTL to kill tumour cells. Here, we describe a proof-of-principle study using a bispecific conjugate designed to retarget ovalbumin (OVA)-specific CTL to kill tumour cells via CD20. A single-chain trimer (SCT) consisting of MHCI H-2K(b)/SIINFEKL peptide/beta 2 microglobulin/BirA was expressed in bacteria, refolded and chemically conjugated to one (1:1; F2) or two (2:1; F3) anti-hCD20 Fab' fragments. In vitro, the [SCT × Fab'] (F2 and F3) redirected SIINFEKL-specific OT-I CTL to kill CD20(+) target cells, and in the presence of CD20(+) target cells to provide crosslinking, they were also able to induce proliferation of OT-I cells. In vivo, activated OT-I CTL could be retargeted to kill [SCT × Fab']-coated B cells from hCD20 transgenic (hCD20 Tg) mice and also EL4 and B16 mouse tumour cells expressing human CD20 (hCD20). Importantly, in a hCD20 Tg mouse model, [SCT × Fab'] administered systemically were able to retarget activated OT-I cells to deplete normal B cells, and their performance matched that of a bispecific antibody (BsAb) comprising anti-CD3 and anti-CD20. [SCT × Fab'] were also active therapeutically in an EL4 tumour model. Furthermore, measurement of serum cytokine levels suggests that [SCT × Fab'] are associated with a lower level of inflammatory cytokine release than the BsAb and so may be advantageous clinically in terms of reduced toxicity.

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More information

Published date: June 2013
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 353481
URI: http://eprints.soton.ac.uk/id/eprint/353481
ISSN: 0340-7004
PURE UUID: 5be01f94-fc02-4dd6-8a73-a4c05d4bc35a
ORCID for Peter W.M. Johnson: ORCID iD orcid.org/0000-0003-2306-4974

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Date deposited: 10 Jun 2013 10:44
Last modified: 15 Mar 2024 02:58

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Contributors

Author: Ben C. King
Author: Angela D. Hamblin
Author: Philip M. Savage
Author: Leon R. Douglas
Author: Ted H. Hansen
Author: Ruth R. French

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