Abstract
Transplant rejection involves a coordinated attack of the innate and the adaptive immune systems of the host. To investigate this dynamic process and the contributions of both donor and host cells, we developed an ear skin graft model suitable for intravital imaging. We found that donor dermal dendritic cells (DCs) migrated rapidly from the graft and were replaced by host CD11b+ mononuclear cells. The infiltrating host cells captured donor antigen, reached the draining lymph node and cross-primed graft-reactive CD8+ T cells. Furthermore, we defined the mechanisms by which host T cells target graft cells. We found that primed T cells entered the graft from the surrounding tissue and localized selectively at the dermis-epidermis junction. Later, CD8+ T cells disseminated throughout the graft and many became arrested. These results provide insights into the antigen presentation pathway and the stepwise progression of CD8+ T cell activity, thereby offering a framework for evaluating how immunotherapy might abrogate the key steps in allograft rejection.
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Acknowledgements
We wish to thank H. Saklani and C. Auriau (Institut Pasteur) for providing B2m−/− mOVA mice and E. Robey and the members of the Bousso laboratory for comments on the manuscript. This work was supported by INSERM, Institut Pasteur and a Marie Curie Excellence grant.
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S.C. designed and carried out the experiments, analyzed the data and wrote the manuscript; M.L.A. developed crucial reagents and participated in experimental design; and P.B. designed the experiments, analyzed the data and wrote the manuscript.
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Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–8 (PDF 1093 kb)
Supplementary Video 1
Mobility of donor dendritic cells in the graft (MOV 2340 kb)
Supplementary Video 2
Donor DCs rapidly die in the draining lymph node (MOV 2012 kb)
Supplementary Video 3
The motility of graft-infiltrating host cells changes over time (MOV 1147 kb)
Supplementary Video 4
Infiltration of the epidermis in the presence of antigenic mismatch (MOV 2506 kb)
Supplementary Video 5
Recipient graft-infiltrating cells are visualized following retransplant (MOV 1091 kb)
Supplementary Video 6
Recipient graft-infiltrating cells can reach the draining lymph node (MOV 2089 kb)
Supplementary Video 7
Skin allografts are efficiently revascularized on day 5 (MOV 3659 kb)
Supplementary Video 8
CD8+ T cells accumulate in the recipient tissue around the graft (MOV 3789 kb)
Supplementary Video 9
CD4+ T cells accumulate in the recipient tissue around the graft (MOV 660 kb)
Supplementary Video 10
CD4+ T cells localize at the dermis/epidermis junction as they enter the graft (MOV 931 kb)
Supplementary Video 11
CD8+ T cells colocalize with propidium iodide positive cells in the allograft (MOV 3328 kb)
Supplementary Video 12
CD8+ T cells colocalize with propidium iodide positive cells in the allograft (MOV 4226 kb)
Supplementary Video 13
CD8+ T cells colocalize with propidium iodide positive cells in the allograft (MOV 686 kb)
Supplementary Video 14
CD8+ T cells colocalize with propidium iodide positive cells in the allograft (MOV 1068 kb)
Supplementary Video 15
CD8+ T cells displayed reduced motility and increased confinement in the allograft (MOV 3723 kb)
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Celli, S., Albert, M. & Bousso, P. Visualizing the innate and adaptive immune responses underlying allograft rejection by two-photon microscopy. Nat Med 17, 744–749 (2011). https://doi.org/10.1038/nm.2376
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DOI: https://doi.org/10.1038/nm.2376
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