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Lymphocytes are subsets of white blood cells protecting us from pathogens and cancer. Lymphocytes perform immune functions by complex and dynamic cell-cell interactions, thus their functions should be understood under right spatio-temporal context. However, studying such complexity and dynamics has been challenging with conventional biological tools. To overcome limitations of conventional approaches, our lab utilize microfabrication, which permits us to recapitulate microenvironments governing molecular/cellular dynamics of lymphocytes. In addition, microfabrication can be used to engineer cellular environments optimal for live-cell fluorescence imaging and single cell-based function analysis. In this presentation, I am going to present how microfabrication and fluorescence live-cell imaging techniques can be combined to investigate lymphocyte functions, in particular tumor cytolysis, which is an essential function for cancer immunotherapy.
Immune system is composed of multiple cells with distinct functions, and immune responses are orchestrated by complex and dynamic cell-cell interactions. Therefore, each cell behavior and function should be understood under right spatio-temporal context. Studying such complexity and dynamics has been challenging with conventional biological tools. Recent development of new technologies such as state of art imaging instruments and microfabrication techniques compatible with biological systems have provided many exciting opportunities to dissect complex and dynamic immune cell interactions; new microscopy techniques enable us to observe stunning dynamics of immune system in real time. Microfabrication permits us to manipulate microenvironments governing molecular/cellular dynamics of immune cells to study detailed mechanisms of phenomena observed by microscopy. Also, microfabrication can be used to engineer microenvironments optimal for specific imaging techniques. In this presentation, I am going to present an example of how these two techniques can be combined to tackle challenging problems in immunology. Obviously, this strategy can readily be applied to many different fields of biology other than immunology.
Cancer immunotherapy has recently been successful in the treatment of various types of tumors. Cytotoxic lymphocytes, including cytotoxic T lymphocytes (CTLs), natural killer (NK) cells, play an essential role in elimination of tumors by directly killing tumor cells. Therefore, evaluation of lymphocyte cytotoxicity against tumor cells is critical for the improvement of cancer immunotherapy. Lymphocyte cytotoxicity is a strictly regulated function requiring a multi-step “checkpoint” to minimize normal cell damage. However, current cytotoxicity assays mostly provide information about final outcomes of cytotoxicity. To overcome this limitation, we are developing new assays that allow “stepwise” evaluation of lymphocyte cytotoxicity using dynamic imaging and microfabrication techniques.