An immune-competent lung-on-a-chip for modelling the human severe influenza infection response

Authors:
Rachel Ringquist , Eshant Bhatia , Paramita Chatterjee , Drishti Maniar , Zhou Fang , Page Franz , Liana Kramer , Delta Ghoshal , Neha Sonthi , Emma Downey , Joshua Canlas , Abigail Ochal , Savi Agarwal , Valeria Cuéllar , Grace Harrigan , Ahmet F Coskun , Ankur Singh , Krishnendu Roy 
In:
Source: Nature
Publication Date: (2025)
Issue: 10: 1038
Research Area:
Immunotherapy / Hematology
Cells used in publication:
Endothelial, umbilical vein, human (HUVEC)
Species: human
Tissue Origin: vein
Fibroblast, lung, human normal (NHLF)
Species: human
Tissue Origin: lung
Epithelial, Small Airway, human (SAEC)
Species: human
Tissue Origin: lung
Experiment

The cell types utilized within the LOC device include endothelial cells, epithelial cells, fibroblasts, macrophages, DCs and white blood cells. The endothelial cells used were primary HUVECs. Specifically, pooled donor HUVECs (Lonza, catalogue number C2519A) were used to minimize donor-to-donor variability. Primary SAECs (Lonza, catalogue number CC-2547S) were used in the epithelial layer of the device. Primary normal human lung fibroblasts (NHLFs; Lonza, catalogue number CC-2512) from two different donors (NHLF-B and NHLF-E) were used in the vascular layer, according to previously performed donor optimization. HUVECs, SAECs and NHLFs were all acquired from Lonza. The macrophages were differentiated from fresh whole blood. 

Abstract

Severe influenza affects 3-5 million people worldwide each year, resulting in more than 300,000 deaths annually. However, standard-of-care antiviral therapeutics have limited effectiveness in these patients. Current preclinical models of severe influenza fail to accurately recapitulate the human immune response to severe viral infection. Here we develop an immune-competent, microvascularized, human lung-on-a-chip device to model the small airways, successfully demonstrating the cytokine storm, immune cell activation, epithelial cell damage, and other cellular- and tissue-level human immune responses to severe H1N1 infection. We find that interleukin-1ß and tumour necrosis factor-a play opposing roles in the initiation and regulation of the cytokine storm associated with severe influenza. Furthermore, we discover the critical stromal-immune CXCL12-CXCR4 interaction and its role in immune response to infection. Our results underscore the importance of stromal cells and immune cells in microphysiological models of severe lung disease, describing a scalable model for severe influenza research. We expect the immune-competent human lung-on-a-chip device to enable critical discoveries in respiratory host-pathogen interactions, therapeutic side effects, vaccine potency evaluation, and crosstalk between systemic and mucosal immunity in human lung.