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The human antifungal immune response primarily relies on neutrophils with contribution from macrophages, while the roles of unconventional lymphocytes such as Natural Killer (NK) cells and mucosal-associated invariant T (MAIT) cells remain less understood. Therefore, using an advanced microfluidic \u201cinvasive aspergillosis-on-chip\u201d (IAC) model, we investigated the coordinated antifungal response of various immune cell types in combination with macrophages within a human tissue-like 3D environment. The IAC model, initially incorporating monocyte-derived macrophages, was enhanced by perfusing either primary neutrophils, NK cells, or MAIT cells for overnight infection experiments. In this model, hyphal growth of A. fumigatus was prevented only when both macrophages and neutrophils were present. However, uncontrolled growth was detected in neutropenic patient scenarios. We observed key interactions such as conidial shuttling and metaforosis between innate immune cells, accompanied by elevated cytokine levels of e.g. IL-1\u03b2 and TNF\u03b1. Importantly, NK cells applied to the vascular compartment of the IAC model significantly reduced hyphal length, confirming their antifungal activity. Similarly, MAIT cells in sufficiently high quantities reduced hyphal growth and branching, although this effect depended on the recognition of riboflavin-derived antigens, as shown by the lack of response to the A. fumigatus \u0394riboB mutant strain. Together, our findings demonstrate that the advanced IAC model is a powerful tool to dissect human antifungal immunity. The interactions observed between macrophages and other immune cells highlight the importance of multi-layered defense strategies against A. fumigatus.<\/jats:p>","DOI":"10.1093\/femsml\/uqag034","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T11:49:22Z","timestamp":1789127362000},"source":"Crossref","is-referenced-by-count":0,"title":["Reconstructing host-pathogen interactions: analysis of the intercellular network of macrophages and other immune cells within an aspergillosis-on-chip model"],"prefix":"10.1093","volume":"7","author":[{"given":"Simranpreet","family":"Kaur","sequence":"first","affiliation":[{"name":"Infections in Hematology and Oncology, Leibniz Institute for Natural Product Research and Infection Biology , 07745 Jena ,","place":["Germany"]},{"name":"Jena Microbial Resource Collection, Leibniz Institute for Natural Product Research and Infection Biology , 07745 Jena 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Biological Sciences, Friedrich Schiller University Jena , 07743 Jena ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4044-9166","authenticated-orcid":false,"given":"Marc Thilo","family":"Figge","sequence":"additional","affiliation":[{"name":"Leibniz Institute for Natural Product Research and Infection Biology Department of Applied Systems Biology, , 07745 Jena ,","place":["Germany"]},{"name":"Institute of Microbiology, Faculty of Biological Sciences, Friedrich Schiller University Jena , 07743 Jena ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"von\u00a0Lilienfeld-Toal","sequence":"additional","affiliation":[{"name":"Infections in Hematology and Oncology, Leibniz Institute for Natural Product Research and Infection Biology , 07745 Jena ,","place":["Germany"]},{"name":"Institute for Diversity Medicine, Faculty of Medicine, Ruhr University Bochum , 44789 Bochum 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