The objective of this study was to determine the role of donor-derived NCMs in the pathogenesis of primary lung allograft dysfunction. Flow cytometry and immunofluorescence microscopy were used to identify NCMs in donor human lungs before and after reperfusion. Murine single-lung transplant was used as a model of human transplantation and to test the effects of donor and recipient treatments, genetic deletion, and cell adoptive transfer. Intravital two-photon microscopy and flow cytometry were used to measure the influx of inflammatory cell populations into the lung. Compartmental staining for flow cytometry, immunofluorescence two-photon microscopy, and immunoelectron microscopy were used to anatomically localize NCMs in the intravascular space. To identify activation pathways and proinflammatory chemokines released by NCMs, RNA sequencing was performed on FACS (fluorescence-activated cell sorter)–sorted donor-derived NCMs in the allograft early after transplant. Genetic deletion of MYD88 and TRIF and measurement of CXCL2 in the posttransplant allograft were performed on the basis of their identification as candidate activation pathways and as an identified neutrophil chemokine during analysis of whole transcriptome data. In an independent set of experiments, to examine the effects of NCMs on the host’s ability to respond to pathogen, mice depleted of NCMs were treated with intratracheal instillation of lipopolysaccharide to model lung injury induced by a Gram-negative organism. In vivo experiments represent pooled results of at least two repeated experiments, unless otherwise indicated. Details of all protocols and primary data for experiments where n < 20 (table S1) are provided in the Supplementary Materials.