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Graft-versus-host disease (GVHD) is a major cause of morbidity and mortality in allogeneic hematopoietic stem cell transplantation (alloSCT). By static microscopy, cutaneous GVHD lesions contain a mix of T cells and myeloid cells. We used 2-photon intravital microscopy to investigate the dynamics of CD4⁺ and CD8⁺ T cells and donor dendritic cells (DCs) in cutaneous GVHD lesions in an MHC-matched, multiple minor histocompatibility antigen-mismatched (miHA) model. The majority of CD4 and CD8 cells were stationary, and few cells entered and stopped or were stopped and left the imaged volumes. CD8 cells made TCR:MHCI-dependent interactions with CD11c⁺ cells, as measured by the durations that CD8 cells contacted MHCI⁺ vs MHCI⁻ DCs. The acute deletion of Langerin⁺CD103⁺ DCs, which were relatively rare, did not affect CD8 cell motility and DC contact times, indicating that Langerin …