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Resource and Technical AdvanceIn-Press PreviewCell biologyInfectious diseaseInflammation Open Access | 10.1172/jci.insight.203042

Macrophages orchestrate antiviral defense and epithelial repair in a human iPSC-derived alveolar air-liquid interface

Declan L. Turner,1 Hannah Baric,1 Katelyn Patatsos,1 Sahel Amoozadeh,1 Michael See,1 Kathleen A. Strumila,1 Jack T. Murphy,1 Jeremy J. Wiyana,1 Liam Gubbels,1 Elizabeth S. Ng,1 Andrew G. Elefanty,1 Melanie R. Neeland,1 Shivanthan Shanthikumar,1 Sarah L. Londrigan,2 Mirana Ramialison,1 Fernando J. Rossello,1 Edouard G. Stanley,3 and Rhiannon B. Werder1

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Turner, D. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Baric, H. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Patatsos, K. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Amoozadeh, S. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by See, M. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Strumila, K. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Murphy, J. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Wiyana, J. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Gubbels, L. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Ng, E. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Elefanty, A. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Neeland, M. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Shanthikumar, S. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Londrigan, S. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Ramialison, M. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Rossello, F. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Stanley, E. in: PubMed | Google Scholar

1Murdoch Children's Research Institute, Melbourne, Australia

2Department of Microbiology and Immunology, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia

3Murdoch Childrens Research Institute, Melbourne, Australia

Find articles by Werder, R. in: PubMed | Google Scholar

Published March 17, 2026 - More info

JCI Insight. https://doi.org/10.1172/jci.insight.203042.
Copyright © 2026, Turner et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published March 17, 2026 - Version history
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Abstract

The lung alveoli are continually exposed to inhaled pathogens and environmental hazards and rely on coordinated communication between alveolar macrophages and type 2 alveolar epithelial cells (AT2s) to maintain homeostasis. Disruption of these interactions can impair immunity and repair, contributing to acute and chronic respiratory diseases. To better define these mechanisms and support therapeutic discovery, we established a human iPSC-derived air-liquid interface platform that captures key features of AT2-macrophage crosstalk. Using this system, we show that coculture enhances AT2-specific transcriptional programs including lipid synthesis, while macrophages actively phagocytose AT2-derived surfactant. iPSC-derived macrophages adopt an alveolar macrophage-like phenotype and respond to AT2-derived M-CSF. During respiratory infection, macrophages played a crucial role in modulating epithelial inflammatory responses, augmenting antiviral immunity, and limiting viral replication. We further identify a previously unrecognized role for macrophages in epithelial repair, where VEGF-mediated signaling to macrophages increases epithelial permeability during viral infection. Together, these findings reveal dimensions of AT2-macrophage cooperation in homeostasis, infection, and repair, and demonstrate how this iPSC-derived platform can be used to dissect mechanisms that may initiate or drive the progression of respiratory diseases.

Graphical Abstract
graphical abstract
Supplemental material

View Supplemental Video 1. Relates to Figure 1. Live cell confocal microscopy taken over 14 hours of iAT2 (marked by SFTPC-tdTomato) and iMacs (stained with CFSE) in coculture at ALI.

View Supplementary Table 1 - custom RSV Flex probe sequences

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