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Visualizing the innate and adaptive immune responses underlying allograft rejection by two-photon microscopy

Nature Medicine volume 17, pages 744–749 (2011)Cite this article

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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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Figure 1: Donor Langerhans cells but not dermal DCs persist in the graft.
Figure 2: Spatiotemporal pattern of graft infiltration by host cells.
Figure 3: Recipient graft-infiltrating cells reach the dLN and cross-prime CD8+ T cells.
Figure 4: Priming and graft localization of CD8+ T cells in the early phase of rejection.
Figure 5: CD8+ T cell dynamics and cytotoxic activity in the graft.

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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.

Author information

Authors and Affiliations

  1. Département d'Immunologie, Institut Pasteur, Unité des Dynamiques des Réponses Immunes, Paris, France

    Susanna Celli & Philippe Bousso

  2. Institut National de la Santé et de la Recherche Médicale (INSERM) U668, Paris, France

    Susanna Celli & Philippe Bousso

  3. Institut Pasteur, Unité d'Immunobiologie des Cellules Dendritiques, Paris, France

    Matthew L Albert

  4. INSERM U818, Paris, France

    Matthew L Albert

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  1. Susanna Celli
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Contributions

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.

Corresponding author

Correspondence to Philippe Bousso.

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The authors declare no competing financial interests.

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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