Abstract:Holotomographic microscopy (HTM) measures the refractive index (RI) tomograms of living cells and tissues in three dimensions. The ability to observe biological processes at high spatial and temporal resolution opens uncharted territories for cell biologists, however, current HTM devices have a limited throughput. We show here the first automated multi-well plate-compatible HTM device, the CX-A. Thanks to state-of-the-art environment control and a new type of autofocus, the CX-A can record multiple conditions in parallel over large fields of view, while its software EVE supports automated single-cell segmentation and quantification. This opens the door to new applications for HTM, from drug screening to systems biology.
Vγ9Vδ2 T cells are potent but elusive cytotoxic effectors. Means to stimulate their function could lead to powerful new cancer immunotherapies. BTN2A1, a surface protein has recently been shown to bind the Vγ9 chain of the γδ TCR but its precise role in modulating Vγ9Vδ2 T cells functions remains unknown. Here we show that 107G3B5, a monoclonal anti-BTN2A1 agonist antibody, significantly enhances Vγ9Vδ2 T cell functions against hematological or solid cell lines and against primary cells from adult acute lymphoblastic leukemia patients. New computer vision strategies applied to holotomographic microscopy videos show that 107G3B5 enhances the interaction between Vγ9Vδ2 T cells and target cells in a quantitative and qualitative manner. In addition, we provide evidence that Vγ9Vδ2 T cells activated by 107G3B5 induce caspase 3/7 activation in tumor cells, thereby triggering their death by pyroptosis. We thus demonstrate that targeting BTN2A1 with 107G3B5 enhances the Vγ9Vδ2 T cell antitumor response by triggering the pyroptosis-induced immunogenic cell death.
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