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Complement-mediated lysis by anti-CD20 mAb correlates with segregation into lipid rafts

Complement-mediated lysis by anti-CD20 mAb correlates with segregation into lipid rafts
Complement-mediated lysis by anti-CD20 mAb correlates with segregation into lipid rafts
Despite the clinical success of anti-CD20 monoclonal antibody (mAb) in the treatment of lymphoma, there remains considerable uncertainty about its mechanism of action. Here we show that the ability of mAbs to translocate CD20 into low-density, detergent-insoluble membrane rafts appears to control how effectively they mediate complement lysis of lymphoma cells. In vitro studies using a panel of anti-B-cell mAbs revealed that the anti-CD20 mAbs, with one exception (B1), are unusually effective at recruiting human complement. Differences in complement recruitment could not be explained by the level of mAb binding or isotype but did correlate with the redistribution of CD20 in the cell membrane following mAb ligation. Membrane fractionation confirmed that B1, unlike 1F5 and rituximab, was unable to translocate CD20 into lipid rafts. In addition, we were able to drive B1 and a range of other anti-B-cell mAbs into a detergent-insoluble fraction of the cell by hyper-cross-linking with an F(ab')(2) anti-Ig Ab, a treatment that also conferred the ability to activate lytic complement. Thus, we have shown that an important mAb effector function appears to be controlled by movement of the target molecule into membrane rafts, either because a raft location favors complement activation by mAbs or because rafts are more sensitive to complement penetration.
0006-4971
1045-1052
Cragg, Mark S.
ec97f80e-f3c8-49b7-a960-20dff648b78c
Morgan, Suzanne M.
e8af487a-f03a-43a5-a0f4-7093b508434b
Chan, H.T. Claude
b109c93f-7e9a-44ee-ad12-da757b1b11fc
Morgan, B. Paul
01432538-99b2-4d74-8e94-bfea3b6c8b7b
Filatov, A.V.
10d2d436-dd82-462e-85eb-70866baa8af2
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
French, Ruth R.
a95ea7a1-7aeb-4c20-998e-fde663613fd1
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded
Cragg, Mark S.
ec97f80e-f3c8-49b7-a960-20dff648b78c
Morgan, Suzanne M.
e8af487a-f03a-43a5-a0f4-7093b508434b
Chan, H.T. Claude
b109c93f-7e9a-44ee-ad12-da757b1b11fc
Morgan, B. Paul
01432538-99b2-4d74-8e94-bfea3b6c8b7b
Filatov, A.V.
10d2d436-dd82-462e-85eb-70866baa8af2
Johnson, Peter W.M.
3f6068ce-171e-4c2c-aca9-dc9b6a37413f
French, Ruth R.
a95ea7a1-7aeb-4c20-998e-fde663613fd1
Glennie, Martin J.
9f6f0eff-4560-48c2-80cd-0ec116110ded

Cragg, Mark S., Morgan, Suzanne M., Chan, H.T. Claude, Morgan, B. Paul, Filatov, A.V., Johnson, Peter W.M., French, Ruth R. and Glennie, Martin J. (2003) Complement-mediated lysis by anti-CD20 mAb correlates with segregation into lipid rafts. Blood, 101 (3), 1045-1052. (doi:10.1182/blood-2002-06-1761).

Record type: Article

Abstract

Despite the clinical success of anti-CD20 monoclonal antibody (mAb) in the treatment of lymphoma, there remains considerable uncertainty about its mechanism of action. Here we show that the ability of mAbs to translocate CD20 into low-density, detergent-insoluble membrane rafts appears to control how effectively they mediate complement lysis of lymphoma cells. In vitro studies using a panel of anti-B-cell mAbs revealed that the anti-CD20 mAbs, with one exception (B1), are unusually effective at recruiting human complement. Differences in complement recruitment could not be explained by the level of mAb binding or isotype but did correlate with the redistribution of CD20 in the cell membrane following mAb ligation. Membrane fractionation confirmed that B1, unlike 1F5 and rituximab, was unable to translocate CD20 into lipid rafts. In addition, we were able to drive B1 and a range of other anti-B-cell mAbs into a detergent-insoluble fraction of the cell by hyper-cross-linking with an F(ab')(2) anti-Ig Ab, a treatment that also conferred the ability to activate lytic complement. Thus, we have shown that an important mAb effector function appears to be controlled by movement of the target molecule into membrane rafts, either because a raft location favors complement activation by mAbs or because rafts are more sensitive to complement penetration.

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More information

Published date: 2003

Identifiers

Local EPrints ID: 26259
URI: https://eprints.soton.ac.uk/id/eprint/26259
ISSN: 0006-4971
PURE UUID: edcb535a-8382-44e0-bd1f-afe34e4802a7
ORCID for Mark S. Cragg: ORCID iD orcid.org/0000-0003-2077-089X
ORCID for Peter W.M. Johnson: ORCID iD orcid.org/0000-0003-2306-4974

Catalogue record

Date deposited: 21 Apr 2006
Last modified: 07 Aug 2019 00:48

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