The immunological drivers of chronic lung allograft dysfunction (CLAD), the major barrier to long-term survival after lung transplantation, are poorly understood at a tissue level. Tissue imaging using mass spectrometry with laser ablation of regions of interest offers single-cell resolution of distinct immune cell populations and their spatial relationships and may improve our understanding of CLAD pathophysiology.Lung tissue from 23 lung transplant recipients, 20 with and 3 without CLAD, was sectioned and stained with a 40-plex antibody panel before 81 regions of interest from airways, blood vessels and lung parenchyma were laser ablated.190,851 individual segmented cells across 41 mm2 tissue were captured before 26 distinct immune and structural cell populations were identified and interrogated across CLAD phenotypes. CLAD was associated with expansion of cytotoxic T cells, γδ T cells and plasma cells and M2 macrophage polarization compared with non-CLAD. Within CLAD, bronchiolitis obliterans syndrome was characterized by more γδ T cells and fewer Th1 cells than restrictive allograft syndrome. Both adaptive and innate immune cells were involved in the temporal evolution of fibrotic remodeling. Although fibrosis seemed to be partially associated with different factors in restrictive allograft syndrome (M2 macrophages, Th1 cells) and in bronchiolitis obliterans syndrome (γδ T cells).Imaging mass cytometry enables in-depth analyses of immune cell phenotypes in their local microenvironment. Using this approach, we identified major differences in cell populations in CLAD versus non-CLAD and in BOS versus RAS, with novel insights into the fibrotic progression of CLAD.