Epitope characterization and crystal structure of GA101 provide insights into the molecular basis for type I/II distinction of CD20 antibodies
2011; Elsevier BV; Volume: 118; Issue: 2 Linguagem: Inglês
10.1182/blood-2010-09-305847
ISSN1528-0020
AutoresGerhard Niederfellner, Alfred Lammens, Olaf Mundigl, Guy Georges, Wolfgang Schaefer, Manfred Schwaiger, Andreas G. Franke, Kornelius Wiechmann, Stefan Jenewein, Jerry W. Slootstra, Peter Timmerman, Annika Brännström, Frida Lindstrom, Ekkehard Mössner, Pablo Umaña, Karl‐Peter Hopfner, Christian Klein,
Tópico(s)T-cell and B-cell Immunology
ResumoCD20 is a cell-surface marker of normal and malignant B cells. Rituximab, a monoclonal antibody targeting CD20, has improved the treatment of malignant lymphomas. Therapeutic CD20 antibodies are classified as either type I or II based on different mechanisms of killing malignant B cells. To reveal the molecular basis of this distinction, we fine-mapped the epitopes recognized by both types. We also determined the first X-ray structure of a type II antibody by crystallizing the obinutuzumab (GA101) Fab fragment alone and in complex with a CD20 cyclopeptide. Despite recognizing an overlapping epitope, GA101 binds CD20 in a completely different orientation than type I antibodies. Moreover, the elbow angle of GA101 is almost 30° wider than in type I antibodies, potentially resulting in different spatial arrangements of 2 CD20 molecules bound to a single GA101 or rituximab molecule. Using protein tomography, different CD20 complexes were found to be associated with the 2 antibodies, and confocal microscopy showed different membrane compartmentalization of these subpopulations of the cellular CD20 pool. Our findings offer a possible molecular explanation for the different cellular responses elicited by type I and II antibodies.
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