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Rationale: Tissue-resident memory T cells (T RM ) play a critical role in the defense against inhaled pathogens. The isolation and study of human lung tissue-resident memory T cells and lung-resident macrophages (M LR ) are limited by experimental constraints. Objectives: To characterize the spatial and functional relationship between M LR and human lung tissue-resident memory T cells using ex vivo lung perfusion (EVLP). Methods: T RM and M LR were isolated using EVLP and intraperfusate-labeled CD45 antibody. Cells isolated after 6 hours of EVLP were analyzed using spectral flow cytometry. Spatial relationships between CD3 + and CD68 + cells were explored with multiplexed immunohistochemistry. Functional relationships were determined by using coculture and T-cell-receptor complex signal transduction. Measurements and Main Results: Lungs from 8 research-consenting organ donors underwent EVLP for 6 hours. We show that human lung T RM and M LR colocalize within the human lung, preferentially around the airways. Furthermore, we found that human lung CD8 + T RM are composed of two functionally distinct populations on the basis of PD1 (programed cell death receptor 1) and ZNF683 (HOBIT) protein expression. We show that M LR provide costimulatory signaling to PD1 hi CD4 + and CD8 + lung T RM, , augmenting the effector cytokine production and degranulation of T RM . Conclusions: EVLP provides an innovative technique to study resident immune populations in humans. Human M LR colocalize with and provide costimulation signaling to T RM , augmenting their effector function.

Human Lung-Resident Macrophages Colocalize with and Provide Costimulation to PD1hiTissue-Resident Memory T Cells

Human Lung-Resident Macrophages Colocalize with and Provide Costimulation to PD1^hiTissue-Resident Memory T Cells