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bioRxiv · 10.1101/2023.02.23.529674

Proteomic profiling of isolated immune synapses from primary mouse B cells

Cunha, D. M.·Hernandez-Perez, S.·Awoniyi, L. O.·Carisey, A. F.·Jacquemet, G.·Mattila, P.
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Abstract

The immune synapse (IS) is a cell-cell interaction platform critical in lymphocyte activation by specific antigens. Despite of B cells being able to also respond to soluble antigens, in particular the in vivo importance of the IS and surface-tethered antigen recognition has strongly emerged in the recent years. The IS serves as a dynamic hub for multiple cellular actions but the molecular details of these functions, especially beyond the B cell antigen receptor (BCR) signalling, remain poorly understood. Here, to address the lack in the systems level understanding of the IS, we setup methodology for comprehensive investigation of the composition of the primary mouse B cells IS at proteome level. Utilizing functionalized magnetic beads to mimic antigen presenting cells and trigger IS formation on them, we developed a method to specifically and robustly extract the cell adhesions on the beads, namely the IS or transferrin receptor mediated adhesion as a control. Our data revealed 661 proteins exclusively present in the IS at 15 minutes after BCR engagement, 13 exclusively in the control adhesions and 365 proteins shared between the samples. We got strong coverage of the known components of the IS as well as identified a plethora of unknown proteins and functional pathways with hitherto unknown roles in B cell IS. Thus, in this work, we validated the IS isolation method as a valuable tool to study early B cell activation by surface-bound antigens as well as unveil several novel proteins and pathways suggestive of new functional aspects in the IS.

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Cunha, D. M., Hernandez-Perez, S., Awoniyi, L. O., Carisey, A. F., Jacquemet, G., Mattila, P.. 2023-02-23. Proteomic profiling of isolated immune synapses from primary mouse B cells. https://doi.org/10.1101/2023.02.23.529674

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