NKG2D receptor activation drives primary graft dysfunction severity and poor lung transplantation outcomes
Daniel R. Calabrese, … , Jonathan P. Singer, Mark R. Looney
Daniel R. Calabrese, … , Jonathan P. Singer, Mark R. Looney
Published November 8, 2022
Citation Information: JCI Insight. 2022;7(24):e164603. https://doi.org/10.1172/jci.insight.164603.
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Research Article Immunology Pulmonology

NKG2D receptor activation drives primary graft dysfunction severity and poor lung transplantation outcomes

Abstract

Clinical outcomes after lung transplantation, a life-saving therapy for patients with end-stage lung diseases, are limited by primary graft dysfunction (PGD). PGD is an early form of acute lung injury with no specific pharmacologic therapies. Here, we present a large multicenter study of plasma and bronchoalveolar lavage (BAL) samples collected on the first posttransplant day, a critical time for investigations of immune pathways related to PGD. We demonstrated that ligands for NKG2D receptors were increased in the BAL from participants who developed severe PGD and were associated with increased time to extubation, prolonged intensive care unit length of stay, and poor peak lung function. Neutrophil extracellular traps (NETs) were increased in PGD and correlated with BAL TNF-α and IFN-γ cytokines. Mechanistically, we found that airway epithelial cell NKG2D ligands were increased following hypoxic challenge. NK cell killing of hypoxic airway epithelial cells was abrogated with NKG2D receptor blockade, and TNF-α and IFN-γ provoked neutrophils to release NETs in culture. These data support an aberrant NK cell/neutrophil axis in human PGD pathogenesis. Early measurement of stress ligands and blockade of the NKG2D receptor hold promise for risk stratification and management of PGD.

Authors

Daniel R. Calabrese, Tasha Tsao, Mélia Magnen, Colin Valet, Ying Gao, Beñat Mallavia, Jennifer J. Tian, Emily A. Aminian, Kristin M. Wang, Avishai Shemesh, Elman B. Punzalan, Aartik Sarma, Carolyn S. Calfee, Stephanie A. Christenson, Charles R. Langelier, Steven R. Hays, Jeffrey A. Golden, Lorriana E. Leard, Mary Ellen Kleinhenz, Nicholas A. Kolaitis, Rupal Shah, Aida Venado, Lewis L. Lanier, John R. Greenland, David M. Sayah, Abbas Ardehali, Jasleen Kukreja, S. Samuel Weigt, John A. Belperio, Jonathan P. Singer, Mark R. Looney

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Figure 5

NKG2D stress ligands mediate killing of airway epithelial cells. (A)

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NKG2D stress ligands mediate killing of airway epithelial cells. (A) Air...
Airway epithelial cells grown in liquid culture were subjected to hypoxia (1% O2) or normoxia (21% O2). (B–D) Transcription at 6 hours (6h), 24 hours (24h), and 48 hours (48h) was measured by PCR for hypoxia inducible factor 1A (HIF1A), MICB, and ULBP4. (E) Representative flow cytometry plots of surface MICB with 24 hours of normoxia or hypoxia. (F) Airway epithelial cell surface expression of MICB. (G) Representative histograms of MICB median fluorescence (MFI). (H) MICB MFI is shown at 6 and 24 hours. (I) To assess NK cell killing of airway epithelial cells, hypoxic or normoxic control cells were cocultured in a 2:1 ratio for 24 hours with primary human NK cells. Hypoxic cells were treated with isotype-matched control antibody or anti-NKG2D blocking antibody for 24 hours preceding the experiment. (J) Dead cell counts are shown at 30-minute intervals across 24 hours comparing total AUC for the 3 conditions. Summary data are displayed with box-and-whisker plots illustrating individual data points, bounded by boxes at 25th and 75th percentiles, and with medians depicted with bisecting lines. Individual P values calculated with Mann Whitney U test (B, C, D, F, and H) and 1-way ANOVA with Tukey’s honestly significant difference post hoc comparisons. ** P < 0.01, *** P < 0.001, **** P < 0.0001.