Imaging dynamics of CD11c+ cells and Foxp3+ cells in progressive autoimmune insulitis in the NOD mouse model of type 1 diabetes
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The aim of this study was to visualise the dynamics and interactions of the cells involved in autoimmune-driven inflammation in type 1 diabetes.
We adopted the anterior chamber of the eye (ACE) transplantation model to perform non-invasive imaging of leucocytes infiltrating the endocrine pancreas during initiation and progression of insulitis in the NOD mouse. Individual, ACE-transplanted islets of Langerhans were longitudinally and repetitively imaged by stereomicroscopy and two-photon microscopy to follow fluorescently labelled leucocyte subsets.
We demonstrate that, in spite of the immune privileged status of the eye, the ACE-transplanted islets develop infiltration and beta cell destruction, recapitulating the autoimmune insulitis of the pancreas, and exemplify this by analysing reporter cell populations expressing green fluorescent protein under the Cd11c or Foxp3 promoters. We also provide evidence that differences in morphological appearance of subpopulations of infiltrating leucocytes can be correlated to their distinct dynamic behaviour.
Together, these findings demonstrate that the kinetics and dynamics of these key cellular components of autoimmune diabetes can be elucidated using this imaging platform for single cell resolution, non-invasive and repetitive monitoring of the individual islets of Langerhans during the natural development of autoimmune diabetes.
KeywordsAnimal–mouse Imaging Islet degeneration and damage Islet transplantation
Anterior chamber of the eye
Green fluorescent protein
Lymphatic vessel endothelial hyaluronan receptor 1
Plasmacytoid dendritic cell
Regulatory T cell
Imaging data were collected at the Center for Advanced Bioimaging (CAB) Denmark, University of Copenhagen, and we would like to thank Michael Hansen (CAB, University of Copenhagen, Denmark) for his excellent technical assistance. B6 Rag2 −/− was a kind gift from Fred W. Alt (Boston Children’s Hospital, Boston, MA, USA).
This work was supported by grants from the European Commission (VIBRANT CP-IP 228933-2), the Danish Research Council, Lundbeckfonden and the Swedish Research Council. LH was supported by a PhD fellowship from the Lundbeck foundation.
Duality of interest
P-OB is founder and CEO of the Biotech Company Biocrine AB. He is also a member of the board of that company. EI is a consultant of Biocrine AB.
AS-C, LH and DH contributed to the conception and design of the experiments; AS-C and LH collected the data; and AS-C, LH, EI, NF-P, UD, SG, ÅL, TDH, AS, P-OB and DH contributed to the analysis and interpretation of the data. All authors contributed to the drafting of the article and revising it critically, and gave final approval of the version to be published.
Repeated non-invasive 2-photon imaging of Foxp3-GFP cells. Time-lapse recordings (26 min, 18 min and 30 min) of the same intraocular islet showing graft-infiltrating GFP-labeled Foxp3+ cells at 3 weeks, 4 weeks and 5 weeks post adoptive transfer. Video is shown as maximum projection and blood vessels are shown in red. Time resolution: 25 sec, scale bar: 100 μm. Related images are shown in ESM Fig. 5. (MOV 5692 kb)
Foxp3-GFP cells moving along blood vessels in the pancreatic parenchyma. Time-lapse recording (40 min) of Foxp3-GFP+ cells moving along blood vessels in the pancreatic parenchyma. Video (shown as maximum projection) was cropped to area of interest (x,y) from ESM video 5. Foxp3-GFP+ cells show the typical elongated shape of fast moving cells. It also shows short-term interaction between GFP-labeled cells in the beginning and the end of the video. Blood vessels are visualized in red. (MOV 2495 kb)