BackgroundCheckpoint inhibitor pneumonitis (CIP) is a rare but potentially fatal immune-related adverse event (irAE) that can interrupt immune checkpoint blockade in non-small cell lung cancer (NSCLC). With no validated pretreatment biomarkers and a diagnosis largely made by exclusion, upfront risk stratification is required. Recent advances in artificial intelligence (AI)-driven pathomics have made it feasible to infer tumor immune microenvironment (TIME)-relevant risk states in patients with NSCLC. Accordingly, we leveraged hematoxylin and eosin(H&E)-based digital pathomics combined with clinical variables to interrogate the TIME in patients who developed CIP and to enable pretreatment and early prediction of CIP.MethodsIn this retrospective study, 346 eligible patients from three hospitals were screened consecutively between January 2022 and January 2025. Patients were divided into CIP and non-CIP groups according to whether CIP occurred at a prespecified observation endpoint. We first developed a pathomics model that employed convolutional neural networks (CNNs) combined with multi-instance learning (MIL) to generate predictions at both the patch and whole slide image (WSI) levels on H&E-stained slides. Separately, we constructed a clinical model using logistic regression (LR) to process the structured clinical data accompanying each case. Subsequently, pathological and clinical information were integrated, where modeling was advanced from modality-specific feature learning to cross-modal representation learning, and final predictive modeling was completed. The predictive performance of different models was evaluated using the area under the Receiver Operating Characteristic (ROC) curve and benchmarked against unimodal models and standard ensemble methods.ResultsWhen the models were evaluated across both internal validation and external test datasets, the pathomics model demonstrated noticeably stronger performance than the clinical approach, achieving area under the curve (AUC) scores of 0.916, 0.875(test 1), and 0.843(test 2), respectively, while the clinical model posted more modest results of 0.880, 0.569(test 1), and 0.594(test 2). The most significant outcome, however, emerged from the multimodal fusion model, which produced the strongest results of all, with performance metrics of 0.930, 0.919(test 1), and 0.905(test 2) in the validation and test phases, respectively.ConclusionPretreatment H&E-derived pathomics, integrated with baseline clinical biomarkers, enable accurate prediction of CIP risk in locally advanced or metastatic NSCLC. This framework supports proactive surveillance and individualized immune checkpoint inhibitor (ICI) strategies and provides a scalable route to decode TIME-relevant states from routine pathology.
Since first described by Paul Ehrlich in 1878, mast cells have been mostly viewed as effectors of allergy. It has been only in the past two decades that mast cells have gained recognition for their involvement in other physiological and pathological processes. Mast cells have a widespread distribution and are found predominantly at the interface between the host and the external environment. Mast cell maturation, phenotype and function are a direct consequence of the local microenvironment and have a marked influence on their ability to specifically recognize and respond to various stimuli through the release of an array of biologically active mediators. These features enable mast cells to act as both first responders in harmful situations as well as to respond to changes in their environment by communicating with a variety of other cells implicated in physiological and immunological responses. Therefore, the critical role of mast cells in both innate and adaptive immunity, including immune tolerance, has gained increased prominence. Conversely, mast cell dysfunction has pointed to these cells as the main offenders in several chronic allergic/inflammatory disorders, cancer and autoimmune diseases. This review summarizes the current knowledge of mast cell function in both normal and pathological conditions with regards to their regulation, phenotype and role.
Primary immunodeficiencies (PIDs), more recently renamed inborn errors of immunity (IEIs), are a diverse group of over 550 genetic disorders. They cause clinically apparent immune dysregulation, leading to infections, autoinflammation, autoimmunity, and cancer. Initially, most IEIs were described as Mendelian disorders with complete penetrance, but the community has now shown that, in most IEIs, some individuals harboring disease-causing genotypes display only partial clinical disease, or no disease at all. Thus, most IEIs are actually Mendelian disorders with incomplete penetrance. Despite the frequency of incomplete penetrance in IEIs, the conceptual framework for systematically categorizing and explaining these occurrences remains limited. Here, I expand on four recurrent themes of incomplete penetrance that we have recently proposed: genetic variant quality, epigenetic and genetic modification, environment, and mosaicism. For each of these principles, I review what is known and unknown and propose future experimental approaches to fill the gaps in our knowledge. I focus on IEIs, but these concepts can be generalized to all genetic diseases.
Anette M. Määttä, MD, L. Pekka Malmberg, MD, PhD, Anna S. Pelkonen, MD, PhD
et al.
Background: The role of early airway hyperresponsiveness (AHR) in the subsequent small-airway lung function remains unclear. Objective: We assessed via a prospective follow-up study the small-airway lung function of schoolchildren with early childhood lower-airway symptoms and AHR to methacholine and compared the findings to the measurements of reference children with no previous or current lung diseases. Methods: During 2004-11, we measured atopic markers, lung function, and airway responsiveness to methacholine in 193 symptomatic children <3 years old. In 2016-18, a follow-up sample of 84 schoolchildren and 40 reference children were assessed for atopic parameters, spirometry, and small-airway lung function. Analysis was performed on the basis of early childhood AHR, early childhood atopy (defined as a positive skin prick test result), and exposure to parental smoking reported in a questionnaire. All the results were compared with those of the reference group. Results: Schoolchildren with early childhood lower-airway symptoms and AHR had higher prebronchodilator area under the reactance curve (AX) z score, lower forced expiratory flow at 50% of forced vital capacity (FEF50%) z score, and higher lung clearance index (LCI) 2.5% compared with those without early childhood AHR and reference children. Moreover, AX and FEF50% z scores only partly improved after bronchodilation. Early childhood atopy and exposure to parental smoking were not associated with school-age small-airway dysfunction. Conclusion: AHR in symptomatic young children associated with subsequent persistent small-airway dysfunction. Further studies with larger samples of symptomatic young children are warranted to determine whether this connection predicts the development of asthma or other obstructive pulmonary diseases as the children grow.
Julius E. Chia, Julius E. Chia, Robert P. Rousseau
et al.
IntroductionForkhead box P3 (Foxp3) T regulatory cells are critical for maintaining self-tolerance, immune homeostasis, and regulating the immune system.MethodsWe investigated interleukin-4 receptor alpha (IL-4Rα) signalling on T regulatory cells (Tregs) during Listeria monocytogenes (L. monocytogenes) infection using a mouse model on a BALB/c background, specifically with IL-4Rα knockdown in Tregs (Foxp3creIL-4Rα−/lox).ResultsWe showed an impairment of Treg responses, along with a decreased bacterial burden and diminished tissue pathology in the liver and spleen, which translated into better survival. Mechanistically, we observed an enhancement of the Th1 signature, characterised by increased expression of the T-bet transcription factor and a greater number of effector T cells producing IFN-γ, IL-2 following ex-vivo stimulation with heat-killed L. monocytogenes in Foxp3creIL-4Rα-/lox mice. Furthermore, CD8 T cells from Foxp3creIL-4Rα-/lox mice displayed increased cytotoxicity (Granzyme-B) with higher proliferation capacity (Ki-67), better survival (Bcl-2) with concomitant reduced apoptosis (activated caspase 3). In contrast to L. monocytogenes, Foxp3creIL-4Rα-/lox mice displayed similar bacterial burdens, lung pathology and survival during Mycobacterium tuberculosis (M. tuberculosis) infection, despite increased T cell numbers and IFN-γ, TNF and IL-17 production.ConclusionOur results demonstrated that the diminished IL-4Rα signalling on Foxp3+ T regulatory cells resulted in a loss of their functionality, leading to survival benefits in listeriosis but not in tuberculosis.
Meik Körner, Michael Spohn, Ulrich Schüller
et al.
Immunologic treatment options are uncommon in low-grade gliomas, although such therapies might be beneficial for inoperable and aggressive cases. Knowledge of the immune and stromal cells in low-grade gliomas is highly relevant for such approaches but still needs to be improved. Published gene-expression data from 400 low-grade gliomas and 193 high-grade gliomas were gathered to quantify 10 microenvironment cell populations with a deconvolution method designed explicitly for brain tumors. First, we investigated general differences in the microenvironment of low- and high-grade gliomas. Lower-grade and high-grade tumors cluster together, respectively, and show a general similarity within and distinct differences between these groups, the main difference being a higher infiltration of fibroblasts and T cells in high-grade gliomas. Among the analyzed entities, gangliogliomas and pleomorphic xanthoastrocytomas presented the highest overall immune cell infiltration. Further analyses of the low-grade gliomas presented three distinct microenvironmental signatures of immune cell infiltration, which can be divided into T-cell/dendritic/natural killer cell-, neutrophilic/B lineage/natural killer cell-, and monocytic/vascular/stromal-cell-dominated immune clusters. These clusters correlated with tumor location, age, and histological diagnosis but not with sex or progression-free survival. A survival analysis showed that the prognosis can be predicted from gene expression, clinical data, and a combination of both with a support vector machine and revealed the negative prognostic relevance of vascular markers. Overall, our work shows that low- and high-grade gliomas can be characterized and differentiated by their immune cell infiltration. Low-grade gliomas cluster into three distinct immunologic tumor microenvironments, which may be of further interest for upcoming immunotherapeutic research.
Immunologic diseases. Allergy, Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Epithelial cells function as the primary line of defense against invading pathogens. However, bacterial pathogens possess the ability to compromise this barrier and facilitate the transmigration of bacteria. Nonetheless, the specific molecular mechanism employed by Mycobacterium tuberculosis (M.tb) in this process is not fully understood. Here, we investigated the role of Rv2569c in M.tb translocation by assessing its ability to cleave E-cadherin, a crucial component of cell-cell adhesion junctions that are disrupted during bacterial invasion. By utilizing recombinant Rv2569c expressed in Escherichia coli and subsequently purified through affinity chromatography, we demonstrated that Rv2569c exhibited cell wall-associated serine protease activity. Furthermore, Rv2569c was capable of degrading a range of protein substrates, including casein, fibrinogen, fibronectin, and E-cadherin. We also determined that the optimal conditions for the protease activity of Rv2569c occurred at a temperature of 37°C and a pH of 9.0, in the presence of MgCl2. To investigate the function of Rv2569c in M.tb, a deletion mutant of Rv2569c and its complemented strains were generated and used to infect A549 cells and mice. The results of the A549-cell infection experiments revealed that Rv2569c had the ability to cleave E-cadherin and facilitate the transmigration of M.tb through polarized A549 epithelial cell layers. Furthermore, in vivo infection assays demonstrated that Rv2569c could disrupt E-cadherin, enhance the colonization of M.tb, and induce pathological damage in the lungs of C57BL/6 mice. Collectively, these results strongly suggest that M.tb employs the serine protease Rv2569c to disrupt epithelial defenses and facilitate its systemic dissemination by crossing the epithelial barrier.
BackgroundGlioma, the most prevalent malignant intracranial tumor, poses a significant threat to patients due to its high morbidity and mortality rates, but its prognostic indicators remain inaccurate. Although TRAF-associated NF-kB activator (TANK) interacts and cross-regulates with cytokines and microenvironmental immune cells, it is unclear whether TANK plays a role in the immunologically heterogeneous gliomas.MethodsTANK mRNA expression patterns in public databases were analyzed, and qPCR and IHC were performed in an in-house cohort to confirm the clinical significance of TANK. Then, we systematically evaluated the relationship between TANK expression and immune characteristics in the glioma microenvironment. Additionally, we evaluated the ability of TANK to predict treatment response in glioma. TANK-associated risk scores were developed by LASSO-Cox regression and machine learning, and their prognostic ability was tested.ResultsTANK was specifically overexpressed in glioma and enriched in the malignant phenotype, and its overexpression was related to poor prognosis. The presence of a tumor microenvironment that is immunosuppressive was evident by the negative correlations between TANK expression and immunomodulators, steps in the cancer immunity cycle, and immune checkpoints. Notably, treatment for cancer may be more effective when immunotherapy is combined with anti-TANK therapy. Prognosis could be accurately predicted by the TANK-related risk score.ConclusionsHigh expression of TANK is associated with the malignant phenotype of glioma, as it shapes an immunosuppressive tumor microenvironment. Additionally, TANK can be used as a predictive biomarker for responses to various treatments and prognosis.
Nadia Viborg, Michail Angelos Pavlidis, Marina Barrio-Calvo
et al.
Abstract Recent findings have positioned tumor mutation-derived neoepitopes as attractive targets for cancer immunotherapy. Cancer vaccines that deliver neoepitopes via various vaccine formulations have demonstrated promising preliminary results in patients and animal models. In the presented work, we assessed the ability of plasmid DNA to confer neoepitope immunogenicity and anti-tumor effect in two murine syngeneic cancer models. We demonstrated that neoepitope DNA vaccination led to anti-tumor immunity in the CT26 and B16F10 tumor models, with the long-lasting presence of neoepitope-specific T-cell responses in blood, spleen, and tumors after immunization. We further observed that engagement of both the CD4+ and CD8+ T cell compartments was essential to hamper tumor growth. Additionally, combination therapy with immune checkpoint inhibition provided an additive effect, superior to either monotherapy. DNA vaccination offers a versatile platform that allows the encoding of multiple neoepitopes in a single formulation and is thus a feasible strategy for personalized immunotherapy via neoepitope vaccination.
Immunologic diseases. Allergy, Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Antigen-specific immune tolerance has been a long-standing goal for immunotherapy for the treatment of autoimmune diseases and allergies and for the prevention of allograft rejection and anti-drug antibodies directed against biologic therapies. Nanoparticles have emerged as powerful tools to initiate and modulate immune responses due to their inherent capacity to target antigen-presenting cells (APCs) and deliver coordinated signals that can elicit an antigen-specific immune response. A wide range of strategies have been described to create tolerogenic nanoparticles (tNPs) that fall into three broad categories. One strategy includes tNPs that provide antigen alone to harness natural tolerogenic processes and environments, such as presentation of antigen in the absence of costimulatory signals, oral tolerance, the tolerogenic environment of the liver, and apoptotic cell death. A second strategy includes tNPs that carry antigen and simultaneously target tolerogenic receptors, such as pro-tolerogenic cytokine receptors, aryl hydrocarbon receptor, FAS receptor, and the CD22 inhibitory receptor. A third strategy includes tNPs that carry a payload of tolerogenic pharmacological agents that can “lock” APCs into a developmental or metabolic state that favors tolerogenic presentation of antigens. These diverse strategies have led to the development of tNPs that are capable of inducing antigen-specific immunological tolerance, not just immunosuppression, in animal models. These novel tNP technologies herald a promising approach to specifically prevent and treat unwanted immune reactions in humans. The first tNP, SEL-212, a biodegradable synthetic vaccine particle encapsulating rapamycin, has reached the clinic and is currently in Phase 2 clinical trials.
Circular RNAs (circRNAs) are involved in the regulation of various diseases, including periodontitis. The objective of this study was to analyze the biological role and regulatory mechanism of circ_0066881 in LPS-induced periodontal ligament cells (PDLCs). Circ_0066881, microRNA-144-5p (miR-144-5p) and retinoid acid-related orphan receptor A (RORA) levels were determined using reverse transcription-quantitative PCR (RT-qPCR) assay. Cell viability detection was performed by Cell Counting Kit-8 assay. Cell apoptosis was assessed through flow cytometry and caspase-3 activity assay. The protein analysis was completed via Western blot. Inflammatory cytokines were measured by ELISA. The target interaction was validated by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The level of circ_0066881 was down-regulated in periodontitis tissues. Overexpression of circ_0066881 relieved LPS-induced cell viability inhibition and apoptosis or inflammation promotion in PDLCs. Circ_0066881 could bind to miR-144-5p. The protective function of circ_0066881 was achieved by sponging miR-144-5p in PDLCs. Circ_0066881 acts as a miR-144-5p sponge to mediate the RORA level. Inhibition of miR-144-5p attenuated LPS-induced cell injury via targeting RORA. All these results demonstrated that circ_0066881 partly prevented LPS-evoked cell dysfunction in PDLCs through miR-144-5p-mediated up-regulation of RORA.
In Japan and other Asian countries, increased fat uptake induced by a westernized diet is thought to be associated with an increased incidence of inflammatory bowel disease, colorectal cancer and food allergies; however, the mechanism for this remains unclear. High-fat diet (HFD)-fed mice are common animal models used to examine the effect of fat intake in vivo. HFDs are reported to exacerbate DSS-induced colitis and intestinal tumorigenesis, but the effect of HFDs on the intestines before disease induction is often overlooked. We found that the intestinal and gut-associated lymphoid tissue (GALT) morphology of HFD-fed mice differed from that of standard diet (SD)-fed mice. To clarify the mechanism by which fat intake increases intestinal diseases, we analyzed the morphological and immunological aspects of the intestines of HFD-fed mice as well as the molecular mechanisms and physiology. Feeding an HFD for 3 weeks induced atrophy of the small intestine, colon and GALT and reduced the number of small intestinal intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs). Feeding an HFD for only one day reduced the number of small intestinal (SI)-IELs and SI-LPLs. The effect of feeding a 3-week HFD continued for 2 weeks after returning to the SD. The effect of the HFD on the intestinal immune system was independent of the gut microbes. We hypothesized that the cytotoxicity of the abundant HFD-derived free fatty acids in the intestinal lumen impairs the intestinal immune system. Both saturated and unsaturated free fatty acids were toxic to intestinal T-cells in vitro. Orally administering free fatty acids reduced the number of SI-IELs and LPLs. Using a lipase inhibitor to reduce the luminal free fatty acids attenuated the HFD-induced changes in the intestinal immune system, while using a statin to reduce the serum free fatty acids did not. Thus, HFD-induced free fatty acids damaged the intestines; this effect was termed "intestinal lipotoxicity". Because sustained reduction of SI-LPLs after HFD feeding exacerbated indomethacin-induced small intestinal damage, lipotoxicity to the human intestines incurred by consuming a westernized diet in Japan may increase intestinal diseases such as IBD, colorectal cancer or food allergies.
Abstract Long noncoding RNAs (lncRNAs) have been demonstrated to play important regulatory roles in gene expression, from histone modification to protein stability. However, the functions of most identified lncRNAs are not known. In this study, we investigated the role of an lncRNA called AW112010. The expression of AW112010 was significantly increased in CD4+ T cells from C57BL/6J mice activated in vivo with myelin oligodendrocyte glycoprotein, Staphylococcal enterotoxin B, or in vitro with anti-CD3 anti-CD28 mAbs, thereby demonstrating that activation of T cells leads to induction of AW112010. In contrast, anti-inflammatory cannabinoids such as cannabidiol or δ-9-tetrahydrocannabinol decreased the expression of AW112010 in T cells. Interestingly, the expression of AW112010 was high in in vitro–polarized Th1 and Th17 cells but low in Th2 cells, suggesting that this lncRNA may regulate inflammation. To identify genes that might be regulated by AW112010, we used chromatin isolation by RNA purification, followed by sequencing. This approach demonstrated that AW112010 regulated the transcription of IL-10. Additionally, the level of IL-10 in activated T cells was low when the expression of AW112010 was increased. Use of small interfering RNA to knock down AW112010 expression in activated T cells led to increased IL-10 expression and a decrease in the expression of IFN-γ. Further studies showed that AW112010 interacted with histone demethylase KDM5A, which led to decreased H3K4 methylation in IL-10 gene locus. Together, these studies demonstrate that lncRNA AW112010 promotes the differentiation of inflammatory T cells by suppressing IL-10 expression through histone demethylation.