Studying the biology of cytotoxic T lymphocytes in vivo with a fluorescent granzyme B-mTFP knock-in mouse

eLife

Elmar Krause¹  Claudia Schirra²  Per-Olof Berggren³  Fritz Benseler³  Nils Brose⁴  Jens Rettig⁵  Praneeth Chitirala⁶  Paloma Martzloff⁶  Keerthana Ravichandran⁶  Hsin-Fang Chang⁶  Christiane Harenberg⁷  Trese Leinders-Zufall⁸  Midhat H Abdulreda⁸ 

[1] Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, United States;Department of Ophthalmology, University of Miami Miller School of Medicine, Miami, United States;Department of Surgery, University of Miami Miller School of Medicine, Miami, United States;Diabetes Research Institute Federation, Hollywood, United States;The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden;Cellular Neurophysiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), Saarland University, Homburg, Germany;Department of Molecular Neurobiology, Max-Planck-Institute of Experimental Medicine, Göttingen, Germany;Diabetes Research Institute and Cell Transplant Center, University of Miami Miller School of Medicine, Miami, United States;

关键词: cytotoxic T lymphocytes;    cytotoxic granules;    immune synapse;    exocytosis;    two-photon microscopy;    STED microscopy;   

DOI  :  10.7554/eLife.58065

来源: DOAJ

【 摘 要 】

Understanding T cell function in vivo is of key importance for basic and translational immunology alike. To study T cells in vivo, we developed a new knock-in mouse line, which expresses a fusion protein of granzyme B, a key component of cytotoxic granules involved in T cell-mediated target cell-killing, and monomeric teal fluorescent protein from the endogenous Gzmb locus. Homozygous knock-ins, which are viable and fertile, have cytotoxic T lymphocytes with endogeneously fluorescent cytotoxic granules but wild-type-like killing capacity. Expression of the fluorescent fusion protein allows quantitative analyses of cytotoxic granule maturation, transport and fusion in vitro with super-resolution imaging techniques, and two-photon microscopy in living knock-ins enables the visualization of tissue rejection through individual target cell-killing events in vivo. Thus, the new mouse line is an ideal tool to study cytotoxic T lymphocyte biology and to optimize personalized immunotherapy in cancer treatment.

【 授权许可】

Unknown