title: A novel technique for the in vivo imaging of autoimmune diabetes development in the pancreas by two-photon microscopy.
abstract: Type 1 diabetes (T1D) is characterized by the immune-mediated destruction of beta cells in the pancreas. Little is known about the in vivo dynamic interactions between T cells and beta cells or the kinetic behavior of other immune cell subsets in the pancreatic islets. Utilizing multiphoton microscopy we have designed a technique that allows for the real-time visualization of diabetogenic T cells and dendritic cells in pancreatic islets in a live animal, including their interplay with beta cells and the vasculature. Using a custom designed stage, the pancreas was surgically exposed under live conditions so that imaging of islets under intact blood pressure and oxygen supply became possible. We demonstrate here that this approach allows for the tracking of diabetogenic leukocytes as well as vascularization phenotype of islets and accumulation of dendritic cells in islets during diabetes pathogenesis. This technique should be useful in mapping crucial kinetic events in T1D pathogenesis and in testing the impact of immune based interventions on T cell migration, extravasation and islet destruction.
authors.affiliation: Type 1 Diabetes Center at the La Jolla Institute for Allergy and Immunology, La Jolla, California, United States of America.
authors.forename: Ken
authors.initials: K
authors.lastname: Coppieters
authors.forename: Marianne M
authors.initials: MM
authors.lastname: Martinic
authors.forename: William B
authors.initials: WB
authors.lastname: Kiosses
authors.forename: Natalie
authors.initials: N
authors.lastname: Amirian
authors.forename: Matthias
authors.initials: M
authors.lastname: von Herrath
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D039521:CD11c Antigen / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis* / Q000473:pathology; D003952:Diagnostic Imaging / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D050417:Insulin-Secreting Cells / Q000166:cytology; D007700:Kinetics; D007962:Leukocytes / Q000166:cytology; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008853:Microscopy / Q000379:methods*; D010179:Pancreas / Q000378:metabolism*; D010641:Phenotype; D017785:Photons; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes
chemical_list: D001426:Bacterial Proteins; D039521:CD11c Antigen; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria