Innate lymphoid cells: Dual roles and therapeutic opportunities in breast cancer.

Quiescent ILC1 cells confer protection against MCMV infection during undernutrition.

The immunobiological functions and therapeutic applications of interleukin-17 family in cancer.

Immune cells and functions in patients with restless legs syndrome.

The lung is a structurally and immunologically complex organ, constantly exposed to airborne microbes, allergens, and pollutants. Understanding how diverse pulmonary immune cells respond to these challenges is critical for advancing respiratory disease research and identifying appropriate therapeutic interventions. Flow cytometry remains a cornerstone of immune profiling, and advances in high-parameter spectral cytometry have significantly expanded its analytical capabilities. However, challenges, such as poor tissue dissociation, spectral overlap, and loss of spatial information, can hinder the comprehensive interpretation of the lung environment. To address these limitations, we developed an optimized spectral flow cytometry platform, utilizing 5-laser Cytek Aurora spectral cytometers, for deep immunophenotyping of murine lung tissue. Our approach integrates in vivo CD45 antibody labeling-administered intravenously and oropharyngeally-to distinguish circulating, airway, and interstitial immune populations, preserving spatial context in single-cell suspensions. We utilize complementary 25+ parameter panels targeting myeloid and lymphoid compartments, built on a shared backbone to enable consistent classification across datasets. Refined tissue processing protocols support optimal recovery of representative lung cell populations, and overnight intracellular staining enhances marker resolution. Using this platform, we reliably resolve stromal, endothelial, and epithelial cells alongside immune subsets-including macrophages, monocytes, dendritic cells, eosinophils, neutrophils, T and B cells, innate lymphoid cells (ILCs), and natural killer (NK) cells-subclassified by activation, function, and tissue residency. Validation with an influenza A virus model confirmed expected dynamic immune responses and revealed previously unrecognized populations. This spatially informed approach enables high-resolution interrogation of pulmonary immunity in health and disease. © 2025 Wiley Periodicals LLC. Basic Protocol 1: Differential in vivo labeling for spatial profiling of pulmonary immune cells by spectral cytometry Basic Protocol 2: Preparation of single-cell suspensions from murine lung tissue for spectral cytometric analysis of immune cell populations Basic Protocol 3: Optimized workflow and spectral flow cytometry panels for profiling of pulmonary immune cell populations in a single-cell suspension.

Group 2 innate lymphoid cells (ILC2) initiate pathologic type 2 inflammation in allergic asthma in response to diverse tissue-derived stimuli. However, the molecular mechanisms by which ILC2 cells integrate and respond to environmental signals are unclear. Here, we show in a mouse model that in allergic asthma, mechanistic target of rapamycin complex 1 (mTORC1) activation in lung ILC2cells increases. Genetic ablation of Raptor, an obligatory component of mTORC1 complex, results in reduced IL-5 and IL-13 production in ILC2 cells and protects mice from allergic inflammation. Pharmacological inhibition of mTORC1 by rapamycin suppresses ILC2 activation and ameliorates allergic lung inflammation. Mechanistically, mTORC1 activation upregulates neuromedin U receptor 1 (NMUR1) expression through epigenetic reprogramming, which augments ILC2 activation in response to neuromedin U (NMU). However, our experiments suggest that NMUR1 is not an exclusive mediator of ILC2 activation downstream of mTORC1. In conclusion, our work reveals that in ILC2s, mTORC1 signaling coordinates neuro-immune crosstalk for optimal activation, and highlights mTORC1 as a potential therapeutic target for allergic asthma.

IL-1 family members as regulators of lymphoid type-2 immunity in cancer.

Tuft cells are a rare type of epithelial cells characterized not only by their tuft-like structure but also by the expression of specific genes, including those encoding the transcription factor Pou2f3 and canonical gustatory signaling proteins. However, tuft cells can be heterogeneous in various features, both across different tissues and within the same tissue. Homeostatic tuft cells are generated from stem/progenitor cells; however, their formation and gene expression profiles are regulated epigenetically and in response to changes in their microenvironments. Ectopic formation of tuft cells, their transdifferentiation into other cell types, and dedifferentiation to stem/progenitor cells have also been found in some tissues upon severe injuries. Tuft cells function as chemosensory sentinels and can detect a variety of pathogens such as bacteria, protists, and helminths with their cell surface receptors. Activation of these receptors in turn activates intracellular signaling cascades, leading to the release of output effectors: the cytokine IL-25, the eicosanoids, and the transmitters acetyl choline and ATP, some of which act on group 2 innate lymphoid cells, triggering innate immune responses, or on neighboring epithelial cells to accelerate cilia beating and increase mucus secretion, or on the nerve terminals to initiate neuroimmune responses. Some tuft cells are also critical to inflammation resolution and tissue repair-an important part of the healing and recovery process. Further elucidation of tuft cells' ligands, respective receptors and downstream signaling pathways, and output effectors can provide more insights into these cells' pivotal roles in health and disease.

The skin serves not only as a physical barrier but also as a dynamic immune organ, where immune cells orchestrate tissue morphogenesis, homeostasis, and repair. Recent advances have revealed that immune cells play pivotal roles during skin development, guiding processes such as vascular formation, epidermal stratification, and hair follicle morphogenesis. In this review, we integrate fundamental mechanistic studies based on mouse models with key clinical observations from human diseases to comprehensively elucidate the contributions of critical immune cell populations—including macrophages, Langerhans cells, dendritic cells, mast cells, and innate lymphoid cells—to normal skin development. We then explore how dysregulation of immune cell functions leads to aberrant skin morphogenesis, contributing to congenital disorders, autoimmune-mediated abnormalities, and fibrotic diseases. By integrating insights from developmental immunology and pathology, we highlight how deviations in immune regulation can disrupt skin architecture and function. Understanding these mechanisms provides a foundation for developing targeted strategies to modulate immune pathways for therapeutic skin regeneration. Future studies integrating spatial and single-cell technologies will further refine our knowledge of immune-tissue crosstalk in skin development and disease.

Introduction Thymic epithelial tumors (TETs) are rare malignancies frequently associated with autoimmunity. However, circulating immune biomarkers for patient stratification and disease monitoring remain undefined. Innate lymphoid cells (ILCs) are emerging regulators of tumor immunity, but their role in TETs has not yet been characterized. Methods Peripheral blood samples from 32 patients with histologically confirmed TETs and 20 healthy donors were analyzed by multiparametric flow cytometry to quantify circulating ILC subsets. Serum cytokine concentrations were measured using multiplex immunoassays. Patients were stratified according to histology, disease activity, and presence of autoimmune manifestations. Results TETs displayed a significant expansion of circulating ILCs, mainly driven by an enrichment of ILC1, which was more pronounced in patients with active disease and in those with thymic carcinoma. Serum IL-18 levels were elevated—particularly in thymic carcinoma—and correlated with higher concentrations of type 2 cytokines (IL-4, IL-5, IL-9, IL-13). No concomitant increase in canonical ILC1 effector cytokines, including IFN-γ, was observed, indicating a functional dissociation between ILC1 expansion and their expected cytokine profile. Discussion These findings delineate a distinct systemic immune signature in TETs, characterized by IL-18 upregulation and altered ILC1 dynamics, with potential implications for immune regulation and autoimmunity. Circulating ILC profiling combined with IL-18 measurement may represent a promising approach for patient stratification, disease monitoring, and the development of novel immunomodulatory strategies in TETs.

The nervous and immune systems cooperate to regulate mucosal barrier integrity. Nevertheless, whether enteric neurons establish neuroepithelial interactions to coordinate immunity remains elusive. Here, we identified neuroepithelial interactions that differentially control intestinal type 1 and type 2 immunity. Gut epithelial cells expressed vasoactive intestinal peptide (VIP) receptor 1 (VIPR1), and chemogenetic modulation of enteric VIPergic neurons led to altered epithelial-derived cytokines. Epithelial-intrinsic deletion of Vipr1 resulted in diminished type 1 immunity, including reduced type 1 alarmins and intraepithelial lymphocytes. In contrast, epithelial Vipr1 deficiency led to enhanced type 2 immunity, comprising increased type 2 alarmins, tuft cells and activated group 2 innate lymphoid cells. Disruption of neuroepithelial VIP-VIPR1 interactions resulted in increased susceptibility to invasive bacterial infection, which contrasted with enhanced resistance to parasite infection. Our work identifies a multi-tissue axis that controls type 1 and type 2 immunity, deciphering how neuroepithelial interactions distinctively set gut immunity programs.

Barrier homeostasis relies on a finely tuned interplay between the immune system, epithelial cells and commensal microbiota. Beyond these regulators, the enteric nervous system has recently emerged as a central hub coordinating intestinal immune responses, although its role in epithelial differentiation has remained largely unexplored. Here, we identify a neuroepithelial circuit in which vasoactive intestinal peptide (VIP)-positive enteric neurons act on VIPR1+ epithelial stem cells to restrain both their proliferation and secretory lineage differentiation. Disruption of this pathway leads to an expansion of tuft cells, enhanced interleukin (IL)-25 production, activation of group 2 innate lymphoid cells (ILC2s) and induction of a type 2 immune response resembling worm expulsion. This phenotype occurs independently of the microbiota but is modulated by the IL-25R-ILC2-IL-13 axis and dietary solid food intake. Our findings expose the enteric nervous system as a critical regulator of epithelial fate decisions and immune balance, complementing established mechanisms that safeguard barrier integrity and mucosal homeostasis.

Type 2 innate lymphoid cells (ILC2s) contribute to type 2 immunity but have also been associated with multiple inflammatory diseases, including airway inflammation and asthma. We report that beyond its function of degrading poly-ubiquitinylated proteins, the immunoproteasome (i-20S) is required for the proper function of ILC2s by controlling their mitochondrial capacity. We found that 90% of the catalytic β subunits of proteasomes in human ILC2s (hILC2s) are the immuno- (β5i) rather than constitutive (β5c) isoform. Specific, noncovalent, reversible inhibition of i-20S β5i (LMP7) in hILC2s induced ROS production, which inhibited aconitase, leading to altered mitochondrial function and reduced levels of ATP. Reprogramming of metabolic status by an LMP7 inhibitor impaired ILC2 activation, without significant cytotoxicity or preventing their recovery. Hence, the selective inhibition of i-20S in ILC2 cells did not kill them but reversibly depleted their ATP, preventing their activation and cytokine secretion. In mice, proteasome inhibition similarly blocked mitochondrial function and ILC2 activation, preventing airway inflammation in response to IL33 and asthma in response to house dust mites. These findings reveal a previously unappreciated linkage between proteasome blockade, central carbon metabolism, and mitochondrial function and identify a strategy to regulate immune cell metabolism in inflammatory diseases.

IL-17A plays a crucial role in the immune defense against Giardia infection, yet its cellular sources remain incompletely defined. In this study, the site-specific expression and cellular origin of IL-17A were investigated, focusing on both adaptive and innate immune cells in the small intestine, Peyer’s patches and mesenteric lymph nodes of Giardia muris infected C57BL/6 mice. RT-qPCR analyses showed that IL-17A mRNA expression was significantly upregulated in the small intestine, slightly elevated in the Peyer’s patches and unchanged in the mesenteric lymph nodes. Flow cytometry revealed that CD4⁺ T helper cells in the lamina propria of the small intestine are the predominant source of anti-giardial IL-17A. No increase in IL-17A was detected by γδT cells, Tc cells, NK(T) cells, B cells, neutrophils, dendritic cells and innate lymphoid cells. Within the CD3− innate cell population, increased IL-17A production was observed in MHC-II+CD11c+CD11b+/− cells, including a subset of cells expressing typical macrophage markers, namely MHC-II⁺CD11c⁺CD11b⁺CD64⁺F4/80⁺ cells. In T cell-deficient mice, both IL-17A expression and parasite clearance were severely impaired. Our findings demonstrate the importance of the adaptive immunity and simultaneously identify Th cells in the lamina propria as the main source of anti-giardial IL-17A, with a possible supporting role from macrophage-like antigen-presenting cells.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-22662-3.

Group 2 innate lymphoid cells (ILC2s) regulate immunity and tissue repair but are rarely found in the brain. Whether ILC2s can infiltrate the brain from bloodstream and the underlying mechanisms involved remain unclear. While ILC2s have recently been identified as key immunosuppressive players in neuroinflammation, their role in brain tissue repair remains promising but underexplored. Here, using in vivo and in vitro expansion of ILC2s, we demonstrate that ILC2s can enter the brain parenchyma from the blood circulation early after ischemic stroke in a CXCR1-dependent manner. Once in the brain, ILC2s improve long-term recovery of sensory-motor functions by promoting initiation of angiogenesis, namely angiogenic sprouting. Mechanistically, ILC2s produce α-calcitonin gene-related peptide (α-CGRP) to enhance angiogenic sprouting. ILC2s depleted of α-CGRP infiltrate the brain but fail to initiate angiogenesis. Impaired function of CGRP receptors on cerebrovascular endothelial cells abolishes the angiogenic effect of ILC2s. These findings highlight ILC2s as a promising target for promoting therapeutic angiogenesis in stroke recovery.

In the almost two decades since their discovery, innate lymphoid cells (ILCs) have received considerable attention for their roles in immune defense and tissue repair. However, a growing body of evidence now highlights a broader functional spectrum, positioning ILCs as key integrators of environmental cues that support tissue-specific metabolic adaptation. These insights challenge traditional immune-centric paradigms, suggesting that ILCs also contribute to core physiological functions of tissues. In this review, we discuss emerging roles for ILCs in the regulation of energy homeostasis during adulthood, including nutrient sensing, uptake, storage, and utilization. We further examine how these functions may be shaped during the neonatal period, a critical window of developmental transitions that coincides with the establishment of ILC tissue residency. We suggest that ILCs may act as early regulators of postnatal metabolic adaptation, playing a fundamental role in the physiological maturation of tissues after birth.

Neuroinflammation and osteomyelitis in adults with Type 2 diabetes mellitus and peripheral neuropathy without and with foot lesions. What comes first?

Severe asthma is characterized by persistent airway inflammation and structural remodeling, including mucus accumulation, epithelial thickening, and subepithelial fibrosis, which are often refractory to conventional therapies. Group 2 innate lymphoid cells (ILC2s) contribute to these pathological changes by producing large amounts of interleukin-5, interleukin-13, and amphiregulin. Although cell cycle regulators have been implicated in immune cell proliferation, their role in ILC2-driven asthma pathogenesis remains unexplored. Here, we identified the cyclin-dependent kinase (CDK) 4/6-ILC2 axis as a previously unrecognized driver of airway remodeling in severe asthma. Using an ovalbumin (OVA)-induced mouse model of severe asthma, we demonstrated that (1) CDK4+ and CDK6+ cells were elevated by 4.0- and 4.5-fold, respectively, in the lungs; (2) treatment with the CDK4/6 inhibitor palbociclib reduced fibrosis and ILC2 expansion by 77% and 87%, respectively; (3) increased ILC2s in the lungs showed high gene expression levels of CDK4, CDK6, and profibrotic factors, including fibroblast growth factor 2, fibroblast growth factor 23, collagen (COL) 4A2, COL10A1, and COL18A1; (4) thymic stromal lymphopoietin stimulation enhanced CDK4/6 protein expression in ILC2s, leading to their proliferation; and (5) palbociclib significantly inhibited the proliferation of ILC2s, at least in part by suppressing retinoblastoma phosphorylation. These findings establish CDK4/6 as a novel molecular pathway regulating ILC2-mediated airway remodeling and highlight its inhibition as a promising therapeutic approach for severe asthma. SIGNIFICANCE STATEMENT: Although cell cycle regulators have been implicated in immune cell proliferation, their role in group 2 innate lymphoid cell-driven asthma pathogenesis remains unexplored. Here, we identified the cyclin-dependent kinase 4/6-group 2 innate lymphoid cell axis as a previously unrecognized driver of airway remodeling in severe asthma.

Chronic psychological stress-orchestrated glial-ILC3 circuit exacerbates intestinal inflammation and depression.

Dynamic regulation of IL-33 in the obesity pathogenesis: Spatiotemporal specificity and functional paradoxes.