| 研究実績の概要 |
During the second year of the project, we employed the label-free microscopy system that was refined during the first year to further study cell responses and cell-to-cell relations in the context of immune activation.
We first pursued our studies on macrophage activation, where we validated our approach on primary cells, and demonstrated that we can achieve over 90% of accuracy when detecting the activation state of both cell lines (Raw264) and peritoneal macrophages. Furthermore, we could also detect different sub-types of macrophages that originate from different progenitors within primary cell populations.
We then further studied lymphocytes cells and their activation process, by first studied different stimulation routes on cells lines, and could show that we can distinguish the effect of a drug cocktail that bypasses standard signaling processes) and the effect of the exposure to artificial antibody presenting cells (APC), which mimic in vivo stimulation.
We then reproduced these results with primary T cells, and showed that the differences are even larger compared to cell lines, demonstrating the ability of our approach to detect lymphocyte activation, and to distinguish purely chemical stimulation and physiological activation through the creation of an immune synapse on the T cell receptor.
Finally, we obtained preliminary results in cell-to-cell interaction. We developed protocols to create immune synapses between T cells and B cells acting as antigen-presenting cells, and obtained first results with our multimodal system.
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