Clinical and experimental immunology最新文献

{"title":"Comprehensive flow cytometry-based diagnosis of XIAP deficiency.","authors":"Dan Tomomasa, Madoka Nishimura, Ayami Ohya, Kay Tanita, Ryosuke Wakatsuki, Ryohei Watanabe, Satoshi Miyamoto, Akihiro Hoshino, Takahiro Kamiya, Takeshi Isoda, Shuya Kaneko, Masaki Shimizu, Atsushi Hijikata, Katsuhide Eguchi, Masataka Ishimura, Yukako Maeda, Kazushi Izawa, Takaaki Meguro, Kosuke Fujimoto, Etsuko Ishikita-Murayama, Kyogo Suzuki, Eri Okura, Tomoko Uehara, Tomotada Takayama, Satoshi Okada, Masatoshi Takagi, Tomohiro Morio, Rebecca A Marsh, Hirokazu Kanegane","doi":"10.1093/cei/uxaf020","DOIUrl":"https://doi.org/10.1093/cei/uxaf020","url":null,"abstract":"<p><strong>Introduction: </strong>Deficiency of X-linked inhibitor of apoptosis protein (XIAP) is an X-linked recessive inborn error of immunity characterized by abnormal immune responses leading to inflammatory bowel disease and hemophagocytic lymphohistiocytosis. Although XIAP protein expression analysis by flow cytometry (XIAP flow) is commonly used to diagnose XIAP deficiency, certain variants may not affect the protein expression, thereby complicating the diagnostic process. XIAP is crucial for the nucleotide-binding and oligomerization domain 2 (NOD2) signaling pathway.</p><p><strong>Methods: </strong>In this study, we aimed to perform a comprehensive analysis of nine patients diagnosed with XIAP deficiency through genetic testing. In addition to XIAP flow, we employed a previously reported method utilizing muramyl dipeptide (MDP) stimulation, a specific agonist of NOD2, to quantitatively evaluate the downstream tumor necrosis factor-alpha (TNFα) production by flow cytometry in patient monocytes (MDP flow).</p><p><strong>Results: </strong>The median mean fluorescence intensity in healthy controls with XIAP flow was 711 (95% confidence interval (CI), 653 to 815) compared to 195 (95% CI, 161-386) in patients with XIAP deficiency (p < 0.0001). The median percentage of TNFα-producing monocytes in controls with MDP flow was 29.1% (95% CI, 19.6 to 53.7), while in patients it was 0.34% (95% CI, 0.18 to 0.82) (p = 0.0008).</p><p><strong>Results: </strong>The receiver operating characteristic curves demonstrated that both XIAP flow and MDP flow exhibited 100% sensitivity and specificity. Taken together, combining XIAP flow and MDP flow analyses allows for a highly accurate diagnosis. (230/250).</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Type I Interferon Signature: a Quantitative Standardized Method for Clinical Application.","authors":"Alessandra Tesser, Paola Bocca, Massimiliano Ulivi, Alessia Pin, Claudia Pastorino, Davide Cangelosi, Elettra Santori, Enrico Drago, Roberta Caorsi, Fabio Candotti, Marco Gattorno, Alberto Tommasini, Stefano Volpi","doi":"10.1093/cei/uxaf018","DOIUrl":"https://doi.org/10.1093/cei/uxaf018","url":null,"abstract":"<p><p>Type I Interferon (IFN) induced gene expression analysis (\"IFN signature\") is employed to categorize pathological conditions that exhibit type I IFN dysregulation and to direct customized therapeutic strategies. For instance, it is used to differentiate patients with IFN-related inflammation from those with conditions primarily mediated by other cytokines, such as juvenile idiopathic arthritis and periodic fevers. Nevertheless, there is currently no standardized method available for clinical practice, and comparing values at different time points or between centers poses a challenge. In this work we described a standardized method based on the development and validation of a synthetic control to solve the problem of test comparison. Inter-assay and inter-laboratory variability were assessed by multiple repeated analyses within the same laboratory, and between two different laboratories involved in the study. The method has been validated by evaluating the IFN signature of 39 patients with inflammatory disorders known to be related or not to type I IFN (i.e. monogenic interferonopathies, SLE, juvenile dermatomyositis, periodic fevers, juvenile idiopathic arthritis). The proposed method proved to be highly reproducible among centers and able to discriminate among IFN-related or non-IFN-related inflammation. The use of a synthetic control minimized the inter-assay and inter-laboratory variability, and thus facilitate data sharing among centers to improve knowledge of IFN-related inflammation and patient's care.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143699631","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mimics and Challenging Presentations of DADA2.","authors":"Admir Öztürk, Lara Yağcı, Serdal Ugurlu","doi":"10.1093/cei/uxaf017","DOIUrl":"https://doi.org/10.1093/cei/uxaf017","url":null,"abstract":"<p><p>Deficiency of adenosine deaminase 2 (DADA2) has been a challenging diagnosis to make since it was first described in 2014. The disease represents a wide range of phenotypes. Therefore, it may present with various clinical patterns. Throughout the years, several difficult-to-diagnose cases of DADA2 were reported in the literature. Although several studies and reviews were published regarding different phenotypes and manifestations of DADA2, a review of challenging cases with diverse combinations of DADA2 manifestations was needed to integrate the knowledge from the literature into the clinical practice. Immunological, hematologic, autoinflammatory, and adult-onset polyarteritis-nodosa patterns were reported in the literature as cases challenging to diagnose. In this review, we aim to summarize the challenging case reports from the literature, provide an algorithmic approach for these kinds of presentations, and share our perspective and recommendations on the topic. Diagnosing DADA2 on time is a vital issue for preventing fatal and debilitating vascular events with anti-TNF-alpha therapy. Thus, early testing for DADA2 in suspected cases is recommended. Family history and genetic testing of the patient and the first-degree relatives are essential for accurate diagnosis. Thorough systemic examination and imaging might help detect clinically silent findings of vasculitis. Enzymatic activity of ADA2, when available, is also a key diagnostic tool that complements genetic testing and clinical evaluation.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143673461","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pirfenidone alleviates interstitial lung disease in mice by inhibiting neutrophil extracellular trap formation and NLRP3 inflammasome activation.","authors":"Qiyan Su, Yingyue Feng, Jin Guo, Xi Cui, Jiarui Zhu, Jumei Yang, Sigong Zhang","doi":"10.1093/cei/uxaf019","DOIUrl":"https://doi.org/10.1093/cei/uxaf019","url":null,"abstract":"<p><strong>Background: </strong>Idiopathic inflammatory myopathy (IIM) is a progressive autoimmune disease characterized by interstitial lung disease (ILD) with limited therapeutics available. Pirfenidone (PFD), a medication utilized for the treatment of idiopathic pulmonary fibrosis, exhibits notable antioxidant, anti-inflammatory and inhibition of collagen synthesis. This study aims to clarify its efficacy and mechanism in treating IIM-ILD.</p><p><strong>Methods: </strong>A murine myositis-associated interstitial lung disease (MAILD) model was used to assess the therapeutic effect of PFD. The serum levels of IL-1β, IL-6 and TNF-α were detected by ELISA. PFD was utilized to disrupt neutrophil extracellular traps (NETs) formation in vitro, and its inhibitory effect on NETs was assessed through immunohistochemistry of CitH3 and MPO in the lung tissue and the serum cfDNA level in mice. Immunohistochemical and western blot was utilized to examine alterations in epithelial-mesenchymal transition (EMT) and NLRP3 inflammasome markers.</p><p><strong>Results: </strong>PFD treatment inhibited pulmonary inflammation and fibrosis in the MAILD model. PFD intervention reduced NETs formation in vitro. PFD treatment significantly reduce NETs infiltration in the lung tissue and the level of cfDNA in the serum of mice. Additionally, PFD down-regulated EMT and NLRP3-related proteins in vivo. PFD treatment also notably reduced serum levels of IL-1β, IL-6 and TNF-α. After NETs stimulation, A549 cells exhibited EMT and activation of NLRP3 inflammasome. PFD attenuated EMT in A549 cells and suppressed the activation of NLRP3 inflammasome.</p><p><strong>Conclusion: </strong>PFD alleviates ILD in a murine MAILD model by inhibiting NETs formation and NLRP3 inflammasome activation, suggesting that PFD might be a potential therapeutic agent for IIM-ILD.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143673436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In-depth immune profiling of a patient with immunodeficiency, centromeric instability, and facial anomalies syndrome type 2 caused by a novel mutation in ZBTB24.","authors":"Colleen M Roark, Diana Ramírez-Vásquez, Jenniffer Yissel Giron Martinez, Xin Zhen, Alexa N Del Bene, Shannon E Gibson, Megan Dobrose, Natasha B Halasa, Lizbeth Blancas-Galicia, Ruben Martinez-Barricarte","doi":"10.1093/cei/uxaf016","DOIUrl":"10.1093/cei/uxaf016","url":null,"abstract":"<p><p>Immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome is a rare genetic disorder characterized by immunodeficiency and chromosomal instability. Mutations in DNA methylation genes such as DNMT3B (ICF1), ZBTB24 (ICF2), CDCA7 (ICF3), and HELLS (ICF4) cause ICF. ICF2 syndrome has been previously described, yet the extent of its clinical presentation and immunological consequences needs to be further elucidated. We describe a patient with a novel homozygous mutation in ZBTB24 (Q375Hfs*3). While infections with extracellular pathogens are frequent in other reported ICF2 patients, our patient also displays infections by intracellular pathogens. At the molecular level, we showed that the novel mutation results in a truncated ZBTB24 protein that disrupts its function in DNA methylation. We thoroughly characterized the immunological consequences of ZBTB24 deficiency using mass cytometry coupled with state-of-the-art computational methods. Our analysis revealed reduced frequencies of natural killer cells and class-switched memory B cell populations in our patient, along with low levels of the immunoglobulin isotypes IgG4 and IgM. Despite observing normal cell frequencies within the T and myeloid compartments, the clinical presentation of this patient suggests a functional defect in immune cells known to be critical to combat intracellular pathogens. Overall, this study expands the clinical and immunological features of ZBTB24 deficiency and highlights the importance of ZBTB24 to the human immune response.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143656477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FcγRIIB (CD32B) antibodies enhance immune responses through activating FcγRs.","authors":"Alexander P Simpson, Robert J Oldham, Kerry L Cox, Martin C Taylor, Sonya James, Ann L White, Yury Bogdanov, Martin J Glennie, Björn Frendeus, Mark S Cragg, Ali Roghanian","doi":"10.1093/cei/uxaf015","DOIUrl":"https://doi.org/10.1093/cei/uxaf015","url":null,"abstract":"<p><p>Fc receptors (FcR) play a key role in coordinating responses from both the innate and adaptive immune system. The inhibitory Fc gamma receptor (FcγRIIB/CD32B; referred to as FcγRII in mice) restrains the immune response, specifically through regulating immunoglobulin G (IgG) effector functions. FcγRII-deficient mice demonstrate elevated incidence and severity of autoimmunity and increased responses to immunization and infections. To explore the potential of FcγRIIB as a target for augmenting vaccines, we tested the ability of monoclonal antibodies (mAb) against mouse FcγRII and human FcγRIIB to enhance humoral responses in preclinical models. We used wild-type (WT) mice, FcγR-deficient mice, and human FcγRIIB transgenic (Tg) mice with either a functional intracellular domain (hFcγRIIB Tg) or lacking immunoreceptor tyrosine-based inhibitory motif (ITIM) signalling capacity (NoTIM). Targeting mouse and human FcγRIIB with antibodies significantly augmented humoral immune responses against experimental antigens and enhanced tumour clearance in vivo. Surprisingly, mAbs without a functional Fc (N297Q; referred to as Fc-null) lacked efficacy. Similarly, blocking FcγRII in mice lacking activating FcγRs failed to enhance immune responses. Conversely, blocking both signalling-competent and signalling-defective (NoTIM) FcγRIIB in Tg mice with a WT, but not Fc-null, FcγRIIB mAb equally enhanced immunity. These data indicate the redundancy of inhibitory signalling in potentiating immune responses in vivo. Collectively, our data suggest that mAb-targeting of FcγRIIB stabilizes mAb Fc and enhances immune responses via Fc-mediated crosslinking of activating FcγRs, irrespective of the inhibitory function of FcγRIIB. These findings support a strategy to boost immune responses in immunization protocols.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633517","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intratumoral Injection of R848 and Poly(I:C) Synergistically Promoted Antitumour Immune Responses by Reprogramming Macrophage Polarization and Activating DCs in Lung Cancer.","authors":"Shuanghui Chen, Linzhao Li, Haohua Yuan, Huan Gui, Quan Wan, Mengjiao Wang, Hang Lv, Chenglv Wang, Lan Zhu, Yingjie Nie, Xiangyan Zhang","doi":"10.1093/cei/uxae110","DOIUrl":"https://doi.org/10.1093/cei/uxae110","url":null,"abstract":"<p><strong>Introduction: </strong>Immunotherapy has rapidly become a primary treatment option for many lung cancer patients because of its success in treating this prevalent and deadly disease. However, the success of immunotherapy relies on overcoming the immunosuppressive tumour microenvironment, making remodelling this environment a potential strategy for lung cancer therapy. Research suggests that Toll-like receptor (TLR) agonists can impede tumour growth by promoting the conversion of tumour-associated macrophages into an M1-like state or enhancing dendritic cell development. However, there is insufficient research on the combined use of TLR agonists for treating lung cancer.</p><p><strong>Methods: </strong>In this study, we examined how TLR agonists such as resiquimod (R848) and poly(I:C) impact lung cancer treatment when used alone or in combination. In vitro, the regulatory functions and mechanisms of R848 and poly(I:C) were analysed in primary macrophages, RAW264.7 cells, and primary dendritic cells(DCs). Tumour treatment efficacy was assessed in vivo with a Lewis lung carcinoma (LLC) mouse model.</p><p><strong>Results: </strong>The combination of R848+poly(I:C) enhances the transformation of macrophages from the M2 phenotype to the M1 phenotype by increasing inflammatory cytokine levels. The percentage of mature DCs expressing MHC-II+CD11c+ and CD86+ cells was significantly higher in the R848+poly(I:C) group compared with the other groups. Intratumoral injection of the synergistic combination of R848+poly(I:C) suppressed tumour growth by increasing the M1:M2 ratio in TAMs, activating DCs, and attracting CD4+ and CD8+ T cells.</p><p><strong>Conclusion: </strong>R848+poly(I:C) synergistically induce M1-like polarization of macrophages, activate DCs, and promote effective antitumour immunity in mice with subcutaneous LLC tumours.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613755","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unveiling WHIM syndrome: Mavorixafor's Emerging Role in Immune Restoration and Therapy.","authors":"Muhammad Sohaib Khan, Bismah Azeem, Ashir Kanwal, Ifra Eeman Ahmed, Anum Zehra, Aqsa Kabir, Waleed Ahmed, Hania Nasir, Momina Khan, Aatika Manzoor, Muhammad Hasanain, Wania Moeen, Muzamil Khan, Gulrayz Ahmed","doi":"10.1093/cei/uxaf014","DOIUrl":"https://doi.org/10.1093/cei/uxaf014","url":null,"abstract":"<p><p>WHIM syndrome is a rare autosomal dominant immunodeficiency disorder and is an abbreviation formed from the initial letters of its main clinical presentations: Warts, Hypogammaglobulinemia, Infections, and Myelokathexis. It stems mainly from mutations where there is a gain of function in the chemokine receptor CXCR4, which is extensively located on leukocytes, and significantly affects the balance of the immune system. Many therapeutic strategies have been widely explored for several years for this immunodeficiency disorder. Mavorixafor, a CXCR4 antagonist, is a recently approved drug by the Food and Drug Administration (FDA) that is being studied for its longer half-life and oral drug route against WHIM syndrome. This review aims to investigate briefly the underlying mechanisms and pathogenesis of WHIM syndrome, and the current effective treatment approaches, for example CXCR4 antagonists or Hematopoietic Stem Cell Transplantation (HSCT), against it. The review also aims to thoroughly assess the efficacy and safety of Mavorixafor in managing WHIM syndrome, exploring its pharmacokinetics, pharmacodynamics, dosing regimens, and safety. Finally, we also investigate important additional therapeutic uses of Mavorixafor.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143596428","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic changes of host immune response during Helicobacter pylori-induced gastric cancer development.","authors":"Weiwei Fu, Xiurui Han, Xinyu Hao, Jing Zhang, Hejun Zhang, Chao Ma, Miao Xu, Jing Zhang, Shigang Ding","doi":"10.1093/cei/uxae109","DOIUrl":"https://doi.org/10.1093/cei/uxae109","url":null,"abstract":"<p><strong>Introduction: </strong>Helicobacter pylori infection is the main risk factor for gastric cancer. Chronic inflammation is usually induced by H. pylori infection and is accompanied by inherent immune disorders. However, the dynamic changes in the host immune response associated with the transition from normal to metaplasia, dysplasia, and gastric cancer are largely undefined.</p><p><strong>Method: </strong>We established the H. pylori induced gastric cancer mice model. The gastric mucosa of H. pylori infected mice were subjected to RNA-sequencing analysis at different stages. We analyzed systemic immune disturbances in the spleen and changes in serum inflammatory cytokines during gastric cancer development, including gastritis, premalignant lesions (pre-gastric cancer), and gastric cancer stages.</p><p><strong>Results: </strong>RNA-sequencing analysis of the gastric mucosa of H. pylori infected mice highlighted the important role of immune-associated pathways (especially inflammatory pathways) during gastric cancer development. Immune cell proportion analysis revealed the stage-dependent involvement of key immune cell types, including increased Th17 cells in early gastritis and pre-gastric cancer stages and decreased central memory CD4+ and CD8+ T cells during gastric cancer transition. Serum inflammatory cytokine analysis showed that IL-6 and IL-10 levels significantly increased, whereas IL-23 levels decreased during the gastric cancer stage.</p><p><strong>Conclusion: </strong>In summary, we illustrated systemic immune disturbances in the spleen and changes in serum inflammatory cytokines during gastric cancer development. Th17 cells were involved in early gastritis and premalignant processes, while central memory T cells participated in gastric cancer transition. Our findings provide valuable insights into identifying key inflection points and associated biomarkers for the early detection, diagnosis, and treatment of gastric cancer.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143585033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The small molecule inhibitor 3PO is a modulator of neutrophil metabolism, ROS production and NET release.","authors":"Michele Fresneda Alarcon, Genna Ali Abdullah, Andy Nolan, Christina Linford, Maria Martina Meschis, Andrew L Cross, Andrew Sellin, Marie M Phelan, Helen L Wright","doi":"10.1093/cei/uxaf012","DOIUrl":"https://doi.org/10.1093/cei/uxaf012","url":null,"abstract":"<p><p>Neutrophils are key effector leukocytes of the innate immune system and play a pivotal role in defending the host against microbial infections. Recent studies have identified a crucial link between glycolysis and neutrophil cellular functions. Using human neutrophils, we have investigated the intricate relationship between glycolysis, extracellular glucose availability, and the enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), in the regulation of reactive oxygen species (ROS) and neutrophil extracellular trap (NET) production. We have identified that PFKFB3 is elevated in rheumatoid arthritis (RA) neutrophils and that the small molecule PFKFB3 inhibitor 3PO is a key regulator of neutrophil ROS and NET production. 3PO blocked the production of ROS and NETs in a dose-dependent manner in both RA and healthy (HC) neutrophils (p<0.01), and RA neutrophils were more sensitive to lower concentrations of 3PO. Bacterial killing was only partially inhibited by 3PO, and the proportion of live neutrophils after 24h incubation was unchanged. Using NMR metabolomics, we identified that 3PO increases the concentration of lactate, phenylalanine and L-glutamine in neutrophils, as well as significantly decreasing intracellular glutathione (adj. p-value<0.05). We also demonstrated that RA neutrophils produce ROS and NETs in culture conditions which mimic the low glucose environments encountered in RA synovial joints. Our results also suggest 3PO may have molecular targets beyond PFKFB3. By dissecting the intricate interplay between metabolism and neutrophil effector functions, this study advances the understanding of the molecular mechanisms governing pro-inflammatory neutrophil responses and identifies 3PO as a potential therapeutic for conditions characterized by dysregulated neutrophil activation.</p>","PeriodicalId":10268,"journal":{"name":"Clinical and experimental immunology","volume":" ","pages":""},"PeriodicalIF":3.4,"publicationDate":"2025-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143447662","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}