Surface Ig variable domain glycosylation affects autoantigen binding and acts as threshold for human autoreactive B cell activation
Theresa Kissel, Changrong Ge, Lise Hafkenscheid, Joanneke C. Kwekkeboom, Linda M. Slot, Marco Cavallari, Yibo He, Karin A. van Schie, Rochelle D. Vergroesen, Arieke S. B. Kampstra, Sanne Reijm, Gerrie Stoeken‐Rijsbergen, Carolien A. M. Koeleman, Lenard M. Voortman, Laura H. Heitman, Bingze Xu, Ger J.M. Pruijn, Manfred Wuhrer, Theo Rispens, T. Huizinga, Hans Ulrich Scherer, Michael Reth, Rikard Holmdahl, René E. M. Toes
Abstract
The hallmark autoantibodies in rheumatoid arthritis are characterized by variable domain glycans (VDGs). Their abundant occurrence results from the selective introduction of N-linked glycosylation sites during somatic hypermutation, and their presence is predictive for disease development. However, the functional consequences of VDGs on autoreactive B cells remain elusive. Combining crystallography, glycobiology, and functional B cell assays allowed us to dissect key characteristics of VDGs on human B cell biology. Crystal structures showed that VDGs are positioned in the vicinity of the antigen-binding pocket, and dynamic modeling combined with binding assays elucidated their impact on binding. We found that VDG-expressing B cell receptors stay longer on the B cell surface and that VDGs enhance B cell activation. These results provide a rationale on how the acquisition of VDGs might contribute to the breach of tolerance of autoreactive B cells in a major human autoimmune disease.