IL-27 Signaling Protects Against Influenza-Associated Pulmonary Aspergillosis Through Inhibition of Type 2 Immunity and Enhanced Antifungal Immunity

Influenza-associated pulmonary aspergillosis (IAPA) is a severe complication of influenza infection associated with substantial mortality. Influenza disrupts pulmonary host defenses and alters innate immune responses, predisposing patients to invasive fungal infection. Interleukin-27 (IL-27) is an immunoregulatory cytokine with context-dependent antiviral and antifungal effects; however, its role during IAPA remains undefined. A mouse model of IAPA was established by infecting wild-type and IL-27 receptor -deficient (Il27ra-/-) mice with influenza A, followed by Aspergillus fumigatus challenge. IL-27 and IL-27R expression were increased during IAPA. Single-cell RNA sequencing identified monocytes as the primary source of IL-27 and T cells as major IL-27r-expressing cells. Il27ra-/- mice exhibited significantly increased pulmonary fungal and influenza viral burden, enhanced type 2 immune responses characterized by elevated IL-4, IL-5, IL-9, IL-13, eosinophils, Th2 cells, pathogenic Th2 cells, and ILC2s. Despite increased eosinophil abundance, eosinophil-mediated conidial killing was impaired in Il27ra-/- mice. IL-27R deficiency also reduced macrophage abundance and impaired macrophage conidial uptake. Conversely, timed administration of rIL-27 enhanced fungal clearance, improved survival, and increased macrophage conidial uptake and augmented eosinophil killing capacity during IAPA. IL-27 signaling is a protective immunoregulatory cytokine during IAPA that limits pathological type 2 inflammation and enhances antifungal effector function of both eosinophils and macrophages. These findings identify IL-27 as a potential therapeutic in IAPA.