High-throughput genetic testing plays a key role in the diagnosis of genetically determined diseases. Its application has significantly increased the diagnostic efficiency of many rare diseases, including inborn errors of immunity (IEI) and hematological diseases. Most of these patients are managed by pediatricians, immunologists, and hematologists. These specialists are often responsible for selecting the appropriate diagnostic strategy, interpreting the results, and making subsequent therapeutic decisions. In this context, cooperation with clinical geneticists is crucial. This review is intended for physicians of various specialties involved in the care of patients with rare genetically determined immunological and hematological diseases. The authors’ intention was to provide an accessible presentation of genetic testing methods used in the diagnosis of IEI and congenital hematological diseases, together with guidance on selecting the most appropriate diagnostic approach. A diagnostic algorithm based on the patient’s clinical phenotype is proposed, and key principles for interpreting genetic test results are discussed.
Autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy is an emerging autoimmune disorder of the central nervous system (CNS). Its potential association with Epstein-Barr virus (EBV) infection remains unclear, with only a few cases reported following CNS EBV infection. Here, we report a 6-year-old Chinese child who developed autoimmune GFAP astrocytopathy (GFAP-A) after EBV encephalitis. The patient presented with a 6-day history of headache and fever. Cerebrospinal fluid (CSF) analysis indicated viral encephalitis, and metagenomic next-generation sequencing confirmed EBV in the CSF. Brain imaging revealed cortical abnormalities, and elevated GFAP-immunoglobulin G titres were detected in both the CSF and serum. The patient showed marked clinical improvement following treatment with antivirals, corticosteroids and intravenous immunoglobulin. This case highlights the potential role of EBV as a trigger for autoimmune GFAP-A and underscores the importance of considering autoimmune mechanisms in paediatric CNS infections.
Human epididymal protein 4 (HE4), also known as protein four-disulfide core domain 2, is a secretory protein that is highly expressed in epithelial ovarian cancer. HE4 has higher specificity and sensitivity than traditional biomarkers in ovarian cancer, making it an effective marker for monitoring the progression of ovarian cancer. Given the similarities between the pathological processes in cancer and autoimmune diseases (ADs), namely overactivation of immune cells and involvement of inflammatory responses, patients with ADs may have an increased risk of cancer. Accumulating evidence suggests that HE4 is strongly associated with ADs such as systemic lupus erythematosus, Sjögren’s syndrome, rheumatoid arthritis-associated interstitial lung disease, and immunoglobulin A nephropathy. Several studies have reported that HE4 reflects disease severity and has excellent potential as a biomarker for the diagnosis and progression monitoring of ADs. In this article, we provide a detailed overview of the research on the biological function of HE4 in ADs and provide insights into whether HE4 has the potential to serve as a candidate biomarker for chronic inflammatory ADs.
Introduction The study aimed to investigate the mechanism of the Argonaute RISC catalytic component 2 (AGO2)–antioxidant protein 1 (ATOX1) axis in the inflammatory response of sepsis. Material and Methods AGO2 was knocked down by tail vein injection of sh-Ago2 lentivirus, and a sepsis mouse model was established using the cecal ligation and puncture (CLP) method. RAW264.7 cells were transfected with AGO2 or ATOX1 knockdown or overexpression plasmids and treated with lipopolysaccharide (LPS) to construct a cellular sepsis model. Hematoxylin-eosin staining was used to evaluate lung and liver tissue damage in mice. Serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured using biochemical analysis. ELISA was performed to determine the levels of inflammatory cytokines: tumor necrosis factor a (TNF-a), interleukin (IL)-6, and IL-1B. The expression of AGO2, ATOX1, inducible nitric oxide synthase (iNOS), and arginase 1 (Arg1) was analyzed using RT-qPCR and Western blotting. The effect of AGO2 on ATOX1 mRNA stability was assessed using an actinomycin D assay, and the interaction between AGO2 and ATOX1 was analyzed via RIP-qPCR. Results AGO2 and ATOX1 were highly expressed in septic mice. In vitro experiments demonstrated that knockdown of AGO2 or ATOX1 suppressed LPS-induced inflammatory responses and macrophage polarization imbalance. AGO2 promoted M1 macrophage polarization and aggravated inflammatory damage by enhancing the stability of ATOX1 mRNA. In vivo experiments further confirmed that AGO2 knockdown significantly alleviated sepsis-induced lung and liver damage, reduced the inflammatory response, and inhibited ATOX1 expression. Conclusions AGO2 binds to ATOX1 mRNA, promoting ATOX1 expression, inducing macrophage polarization imbalance, and exacerbating sepsis-induced inflammatory responses.
Introduction COVID-19 and venous thrombosis pose significant global health challenges. Recent studies suggest a potential overlap in their underlying molecular mechanisms, particularly immune responses and inflammation. This study aims to elucidate shared molecular pathways between COVID-19 and venous thrombosis, identify common biomarkers, and propose potential therapeutic targets through a comprehensive multi-dataset analysis. Material and Methods Public datasets (GSE171110, GSE189990, GSE178246, GSE48000, and GSE19151) were analyzed using the “Limma” package in R for differential gene expression analysis. Functional enrichment (GO, KEGG) was conducted via the “ClusterProfiler” package. Protein-protein interaction (PPI) networks, transcription factor (TF)-target, and miRNA-target networks were constructed using TRRUST and miRDB databases. Drug sensitivity was assessed using CellMiner, and feature genes were identified using LASSO regression. Immune infiltration analysis was performed with CIBERSORT, and key genes were validated using qRT-PCR and animal models. Results Key differentially expressed genes (DEGs), including SELP and CLEC4D, were identified, highlighting their roles in immune regulation and thrombosis. Drug sensitivity analysis revealed correlations with specific chemotherapeutic agents. Immune infiltration analysis demonstrated increased expression of SELP and CLEC4D in certain immune cells, confirmed by qRT-PCR and in animal models. Conclusions This study uncovered shared molecular mechanisms between COVID-19 and venous thrombosis, identifying potential biomarkers and therapeutic targets such as SELP and CLEC4D. These findings provide new insights into the diagnosis and treatment of both conditions.
The immune-modulatory role of interleukin 10 (IL-10) in breast cancer remains controversial, with reports of both pro- and anti-tumor effects. While B cells are important targets of IL-10, the frequency, phenotype, and functional impact of B cells expressing IL-10 receptor (IL-10R) in tumor-draining lymph nodes (TDLNs) of breast cancer patients are still unclear. In this study, we analyzed axillary lymph nodes from 49 breast cancer patients, assessing IL-10R expression on B cells and the effects of recombinant IL-10 (rIL-10) on granzyme B, CD86, and tumor necrosis factor (TNF-) production. Flow cytometry revealed that 52.9 ±17.3% of B cells expressed IL-10R, predominantly exhibiting an activated/memory phenotype (CD27+CD24hi). Interestingly, IL-10R+ B cells were significantly reduced in metastatic lymph nodes (MLNs) compared to non-metastatic lymph nodes (nMLNs), suggesting a protective role in early disease. Paradoxically, HER2+ tumors exhibited higher frequencies of IL-10R+ B cells, indicating context-dependent functions. Functional assays demonstrated that IL-10 significantly enhanced granzyme B production in B cells but suppressed TNF-hi B cells by approximately 40%, without altering CD86 expression. These findings highlight the dual role of IL-10 in modulating B cell-mediated immunity. IL-10 enhances cytotoxic function via granzyme B while dampening antitumor TNF- responses. The reduced frequency of IL-10R+ B cells in MLNs suggests their potential role in anti-tumor immunity, while their association with HER2+ tumors may indicate a pro-tumor function in certain contexts. This study emphasizes the complexity of IL-10 signaling in B-cell-mediated immunity, shedding light on its multifaceted effects and the need to clarify the role of IL-10 in cancer immunity.
Introduction:Macrophages play a key role in infection, and musculin (MSC) is closely associated with immune cells. The objective of this study was to investigate the effect of MSC on the immune function of mouse peritoneal macrophages. Material and methods:Peritoneal macrophages from wild-type (WT) and MSC knockout (MSC-/-) mice were obtained and cultured for 12 hours. The cells were subsequently treated with lipopolysaccharide (LPS). After 24 hours, the peritoneal macrophages and their supernatants were collected. The changes in the adhesion index in each group were then recorded by cell counting, and the phagocytic index and phagocytic rate were calculated via bacterial phagocytosis experiments. Additionally, the levels of interleukin (IL)-1 β, IL-6, IL-10 and tumour necrosis factor α (TNF-α) in the cell supernatant were measured via enzyme-linked immunosorbent assay (ELISA). The ratios of cluster of differentiation (CD) 16/32 and CD206 were analysed via flow cytometry. Results:At 24 hours after LPS treatment, the number of peritoneal macrophages in the MSC-/- group was lower than that in the WT group, while the adhesion index in the former group was higher than that in the latter group. Compared with those in the WT group, the phagocytosis index and phagocytosis rate were significantly lower in the MSC-/- group, while the levels of IL-1 β, IL-6, and TNF-α were higher, and the level of IL-10 was lower. In addition, the number of type 2 (M2) macrophages was lower in the same comparison. Conclusions:Musculin may enhance the immune function of peritoneal macrophages in mice.
The therapeutic potential of cancer immunotherapy has been increasingly recognized, particularly in strategies that exploit the body’s own immune system to target and eliminate tumor cells. One promising approach involves the use of dendritic cells (DCs) as powerful antigen-presenting cells to generate antigen-specific T cells capable of targeting cancer cells. In this study, we explore the use of placental gp96, a stress-induced protein overexpressed in various tumors, to prime expanded dendritic cells for the generation of antigen-specific T cells. The expanded DCs, loaded with placental gp96, were shown to induce potent anti-tumor immunity in vitro, as evidenced by the strong T cell proliferation, activation, and cytotoxicity against cancer cell lines such as MCF-7, glioblastoma cell line U87MG and the neuroblastoma cell line SH-SY5Y. Notably, placental gp96-loaded DCs induced significantly higher cytotoxicity (67-71%) against tumor cell lines compared to recombinant gp96 (23-26%, p < 0.001), underscoring its clinical potential. This activity may be mediated by various tumor-associated peptides presented on the placental gp96. Moreover, this anti-tumor effect is MHC-restricted. These findings suggest that placental gp96-loaded expanded DCs hold significant promise as a novel immunotherapy strategy for cancers.
Introduction:Mycobacterium tuberculosis-mediated tuberculosis (TB) is an infectious disease that results in approximately 1.2 million deaths annually. Glutaminase (GLS1) is a metabolic enzyme involved in glutaminolysis. The current study examined the roles and mechanisms of GLS1 in TB progression. Material and methods:Peripheral blood mononuclear cells (PBMCs) were extracted from TB patients and healthy individuals, and CD4+ T and CD14+ monocytes were sorted. PBMCs were incubated with M. tuberculosis strain H37Rv lysate to stimulate immune responses and treated with BPTES, a GLS1 inhibitor. RT-qPCR and western blotting were employed to detect mRNA and protein levels, respectively. Immunophenotyping of cells was performed using flow cytometry. ELISA was used to determine cytokine levels. Colony-forming unit assays were used to evaluate M. tuberculosis survival in macrophages. ChIP assays were used to detect the enrichment of H3K9ac/H3K27ac at the gene promoters. Results:GLS1 was elevated in CD4+ T cells from TB patients and H37Rv lysate-stimulated PBMCs. GLS1-mediated glutaminolysis promoted Th1 and Th17 cell differentiation. Inhibition of GLS1 by BPTES facilitated M. tuberculosis survival in macrophages. GLS1 inhibition reduced H3K9ac and H3K27ac epigenetic modification in the promoter region of interferon γ and interleukin 17. Conclusions:GLS1-mediated glutaminolysis may regulate TB progression by modulating Th1 and Th17 immune responses via epigenetic regulation.
Systemic inflammatory response syndrome (SIRS) is recognized to be an exaggerated defense response to various stressors, including trauma. CD4+ T-regulatory cells (CD4+ Tregs) are key mediators in balancing inflammatory processes. Recent findings demonstrated that platelets and CD4+ Tregs interact after injury and SIRS. Therapeutic strategies to modulate the activation of these and other immune cells in SIRS are currently lacking. Ancrod has immunomodulatory effects on CD4+ Tregs that could be beneficial in the treatment of SIRS. Therefore, we studied the impact of ancrod on activation levels of CD4+ Tregs and CD4+ non-Tregs, platelets, and antigen-presenting cells (APC) in vitro and in vivo. We tested the in vitro effect of ancrod (0-15 IU) and the in vivo effect of 8 IU ancrod vs. saline. After collection of spleens and platelet-rich plasma of male C57Bl/6N mice, cells were isolated and incubated with ancrod for 2 hours. Furthermore, we tested the effect of stimulation. CD4+ Tregs, CD4+ non-Tregs, APC and platelet activation were analyzed by flow cytometry. Our results demonstrate that ancrod exerts selective effects on different cell populations. Ancrod affects the adaptive and innate immune responses. Furthermore, we identified a differential effect of ancrod on CD4+ Tregs versus CD4+ non-Tregs depending on the mode of cellular stimulation. Additionally, our findings suggest a dose-dependent role of ancrod in the modulation of platelet activation. Our findings provide the first evidence supporting the potential of ancrod in selectively modulating immune cells, highlighting ancrod as a promising candidate for further investigation.
Chronic myelomonocytic leukemia (CMML) is a myelodysplastic/ myeloproliferative neoplasm. In 2022, updated diagnostic classifications for CMML were introduced by the International Consensus Classification (ICC) and World Health Organization (WHO). Monocytes are subdivided into three populations: classical (MO1), intermediate (MO2), and non-classical (MO3). One of the newly established diagnostic criteria for CMML is an increase in the MO1 fraction to ≥ 94%, as determined by multiparametric flow cytometry (MFC). This parameter has been shown to be a highly sensitive and specific marker that can rapidly and accurately differentiate CMML from other conditions. At the Department of Hematology, Cellular Therapies, and Internal Medicine, University Clinical Hospital in Wrocław, we evaluated the distribution of monocytes and their subpopulations using MFC in 27 patients with newly diagnosed CMML, classified according to the updated WHO criteria. The criterion of an MO1 fraction ≥ 94% was fulfilled in 22 patients (81.5%). Our findings are consistent with previously published data and support the utility of this method as a reliable tool for both initial screening and longitudinal monitoring of CMML.
Introduction:A few studies have reported that 22q11.2 del syndrome (DiGeorge syndrome - DGS) is associated with alterations in B-lymphocytes and innate immunity that predispose to infections and autoimmunity. The present study investigated the B-cell compartment, autoimmunity markers, and components of innate immunity, including NK cells, NK-specific cytotoxicity, phagocytic functions, and adhesion molecules. Material and methods:Thirty-five DGS patients and twenty healthy controls were evaluated. Nephelometric, flow cytometric, ELISA and immunofluorescence techniques were used. Results:There was no significant difference between the study and control groups regarding gender and age. Serum IgG, IgM levels and percentages, and antibodies against vaccine antigens were significantly lower in DGS patients. In DGS patients, 57.2% had low levels of non-switched memory B-cells, and 22.8% had low values of switched memory B-cells. Immature transitional B-cells, immature B-cells, plasmablasts, and active B-cells were significantly elevated. Autoantibodies were found positive in between 2.9% and 14.3% in the patient group. Natural killer T (NKT) cells and regulatory T cells (Tregs) were significantly decreased, whereas Fas+ active cytotoxic cells and Fas+ naive T helper cells were significantly elevated in DGS patients. NK-specific cytotoxicity was found to be higher in the patient group. Conclusions:Defects in humoral immunity, including immunoglobulin deficiencies and decreases in class-switched and nonclass-switched memory B-cell compartments, were common in DGS patients. While decreased Treg and NKT cells play a role in the increased incidence of autoimmunity, increased Fas+ cells counteract autoimmunity. Based on all these data, we would like to emphasize the importance of monitoring DGS patients for immune dysregulation.
Introduction:Interleukin (IL)-33 is a pleiotropic cytokine in the immune system and inflammatory responses, which is involved in cerebral ischemia/reperfusion (I/R) injury. Evidence indicates that IL-33 plays an essential role in macrophage polarization activation, yet the potential molecular mechanisms remain elusive. This study explored the role of IL-33 in the activation of microglia/macrophage-mediated autophagy for cerebral I/R injury via in vitro experiments. Material and methods:Primary macrophages were harvested from mouse bone marrow, and microglia were isolated from mouse brains. Lipopolysaccharide (LPS) and interferon γ (IFN-γ) induced polarization toward the M1 phenotype macrophage, and IL-4 induced polarization toward the M2 phenotype macrophage. Oxygen glucose deprivation/reoxygenation (OGD/R) models of primary microglia were established and co-cultured with M1 and M2 macrophages, respectively. The IL-33 knockout microglia OGD/R model and M1/M2 macrophages were co-cultured to explore the effects of IL-33 in the transformation from pro-inflammatory (M1) phenotype to anti-inflammatory (M2) phenotype against inflammation and autophagy. Microglial activation/polarization, autophagy, and inflammation were detected using western blot, immunofluorescence, and ELISA. Results:Compared with microglia co-cultured with M1 macrophages, microglia co-cultured with M2 macrophages showed significantly higher expression of M2 intracellular markers (STAT6, PPAR-γ, IRF4, CEBP-β, and CD206 (p < 0.05). In contrast, the expression of STAT1 and IRF8 was significantly lower. IL-33 deficiency enhanced the anti-inflammatory phenotype of microglia/M2 macrophages. Furthermore, the expression of IL-10 and transforming growth factor β (TGF-β) was upregulated (p < 0.05), and the expression of IFN-γ was downregulated (p < 0.05) in microglia/M2 macrophages in comparison with microglia/M1 macrophages. Compared with the OGD/R model group, M1 macrophage co-culture increased Beclin-1 and ULK1 expression and reduced P62 expression, and M2 co-culture as well as IL-33 deletion reduced Beclin-1 and ULK1 expression and increased P62 expression levels. Conclusions:IL-33 deficiency inhibited hypoxia-induced microglia autophagy and promoted macrophage polarization toward the anti-inflammatory phenotype.
Introduction:Rheumatoid arthritis (RA) is a chronic autoimmune disease influenced by environmental factors. Double-stranded DNA (dsDNA) serves as an antigen in various autoimmune diseases and is affected by meteorological factors. However, the relationships among the circulating dsDNA level, RA inflammatory immune mechanisms, and seasonal meteorological factors remain unclear. Material and methods:Serum dsDNA levels were measured in patients with RA and healthy controls, and correlations with archived data on seasonal meteorological changes were examined. Quantitative PCR was performed to assess the gene expression of relevant receptor markers in the peripheral circulation of patients and controls. Results:The mean serum dsDNA level was significantly higher in patients with RA than in controls (1.16 ±0.36 ng/ml vs. 0.92 ±0.20 ng/ml, p < 0.001). The expression of genes related to dsDNA receptors, single-stranded RNA receptors, dsRNA receptors, ALR inflammasome-associated inflammatory factors, interferon (IFN) α1, IFN β1, and low-density lipoprotein was upregulated in patients with RA. Conclusions:dsDNA levels in the peripheral circulation of patients with RA are elevated and affected by seasonal climatic factors. This elevation enhances the expression of ALR inflammasome-related inflammatory factors and type I IFN. These findings provide a foundation for future investigations into the mechanisms underlying the link between climate variability and RA progression.
Sporotrichosis, caused by Sporothrix schenckii, typically infects humans through skin trauma, leading to nodules or ulcers that complicate diagnosis. Rare forms, such as disseminated cutaneous sporotrichosis, occur more often in immunosuppressed patients or those with diabetes or alcohol use disorder. We report a 49-year-old male patient with type 2 diabetes mellitus who developed painful ulcerations on his lower limbs following a cat scratch. A deep tissue biopsy confirmed mycelial hyphae, and culture identified Sporothrix spp. A biopsy from a left thigh lesion revealed syphilitic exanthem findings. Treponema pallidum antibodies were positive, with a TPHA titer of 1 : 320. The patient was diagnosed with sporotrichosis and late latent syphilis, treated with benzathine penicillin for syphilis and itraconazole for sporotrichosis, resulting in significant improvement after two months. This case underscores the importance of mycological diagnostics - deep tissue biopsy, culture, and histopathological analysis - in distinguishing sporotrichosis from similar conditions, especially in immunocompromised patients with unresponsive lesions.
Introduction:Aim of the study was to investigate the roles and interaction mechanisms of RAB40C and SNX9 in prostate adenocarcinoma (PRAD) progression and their impact on the Hippo signaling pathway. PRAD is a significant health concern, and understanding the molecular underpinnings is essential for its effective management. Objective of this study was to identify key genes and pathways involved in PRAD using weighted gene co-expression network analysis (WGCNA) and determine the functional implications of RAB40C and its relationship with SNX9. Material and methods:WGCNA was used to chart gene co-expression patterns in PRAD. Functional enrichment analyses of significant modules were performed, and prognostic insights were derived through differential gene expression analysis. The interaction between RAB40C and SNX9 was elucidated using various in vitro assays and databases. Results:WGCNA identified a module (MEblue) highly correlated with PRAD. The hub gene was revealed, with RAB40C being central to PRAD progression. The knockdown of RAB40C inhibited PRAD cell proliferation, migration, and invasion. SNX9 was identified as a substrate protein interacting with RAB40C. The silencing of RAB40C led to increased SNX9 expression, suggesting an inverse regulatory relationship. RAB40C promoted SNX9 degradation via the ubiquitin-proteasome pathway. Silencing RAB40C reduced PRAD cell proliferation, migration, and invasion, effects that were counteracted by simultaneous SNX9 suppression. The interaction between RAB40C and SNX9 influenced target proteins of the Hippo signaling pathway. Conclusions:RAB40C is essential for the progression of PRAD, partly through modulating SNX9 levels and the Hippo signaling pathway. This interplay offers novel insights for PRAD therapeutic strategies.
Introduction:The core event of gout is intensified macrophage inflammation. Our preliminary research found that carbohydrate antigen 72-4 (CA72-4) secreted by monosodium urate (MSU)-induced human synovial cells (HFLS) could upregulate transforming growth factor β1 (TGF-β1) in macrophages. Herein, the mechanism of MSU-induced abnormal upregulation of CA72-4 in HFLS was investigated. Material and methods:Cell viability was assessed by CCK-8 assay. The secretion levels of cytokines and CA72-4 were measured by ELISA. Methylation-specific PCR (MSP) was employed to detect the methylation level of the Cosmc promoter. The molecular interactions were analyzed by RIP and ChIP assays. Results:Our results demonstrated that CA72-4 derived from MSU-treated HFLS markedly inhibited MSU-induced macrophage inflammation. Mechanistically, MSU-induced lncSLED1 upregulation in HFLS reduced MSU-induced macrophage inflammation by promoting CA72-4 secretion. In addition, lncSLED1 facilitated CA72-4 secretion in MSU-treated HFLS by promoting the methylation level of the Cosmc promoter through recruiting EZH2. As expected, Cosmc silencing in HFLS reversed the weakening effect of lncSLED1 downregulation on mCM (the conventional medium from MSU-treated HFLS)-induced inhibition of MSU-induced macrophage inflammation. Conclusions:MSU elevated lncSLED1 in HFLS, boosting CA72-4 secretion by increasing Cosmc promoter methylation via EZH2 recruitment, thus reducing MSU-induced macrophage inflammation.
Introduction:Acute lung injury (ALI) is characterized by inflammation of the lungs, leading to impaired gas exchange and respiratory distress. This study aimed to investigate the mechanisms underlying the role of Krüppel-like factor 14 (KLF14) in ALI pathogenesis. Material and methods:Protein and gene expression levels were quantified using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. Cell viability and pyroptosis were assessed by Cell Counting Kit-8 (CCK-8) and flow cytometry. Interleukin (IL)-18 and IL-1 β levels were measured by enzyme-linked immunosorbent assay (ELISA), while lactate dehydrogenase (LDH) activity was evaluated with a commercial assay kit. The interaction between Deltex E3 ubiquitin ligase 3-like (DTX3L) and KLF14 or nucleotide-binding domain like receptor 3 (NLRP3) was analyzed using chromatin immunoprecipitation (ChIP), co-immunoprecipitation (Co-IP), and dual-luciferase reporter assay. Results:BEAS-2B cells treated with lipopolysaccharide (LPS) exhibited reduced viability and elevated pyroptosis-related markers. Notably, KLF14 upregulation suppressed NLRP3-mediated pyroptosis in LPS-induced cells. Similarly, DTX3L overexpression attenuated pyroptosis in LPS-treated BEAS-2B cells. Mechanistically, KLF14 enhanced DTX3L transcription, and DTX3L promoted NLRP3 degradation via ubiquitination. Furthermore, KLF14 upregulation inhibited NLRP3-driven pyroptosis by inducing DTX3L expression. Conclusions:Upregulation of KLF14 inhibited NLRP3-mediated pyroptosis through DTX3L activation, thereby improving sepsis-induced acute lung injury.
Chitinase-3-like protein 1 (CHI3L1) is overexpressed in various types of diseases, particularly in sepsis, and contributes to disease progression. However, the role of CHI3L1 and the pathways involved in the progression of sepsis are not yet completely understood. In the present study, lipopolysaccharide (LPS) was used to construct mouse and cell models of sepsis. Reverse transcription-quantitative PCR was used to detect the expression of CHI3L1. Cycloheximide was used to analyze the protein stability of CHI3L1. The mRNA levels of CHI3L1 and the lactate content were found to be significantly elevated in the serum of patients with sepsis and in LPS-exposed RAW264.7 cells. Furthermore, LPS exposure significantly increased the lactylation of CHI3L1 in RAW264.7 cells. Lactate treatment promoted the lactylation of CHI3L1 and 2-DG treatment inhibited it. Lactylation induced the degradation of CHI3L1 protein. Furthermore, CHI3L1 knockdown inhibited macrophage apoptosis and the release of inflammatory cytokines (IL-1 β, IL-6 and TNF-α) and inactivated the NF-κB pathway in the LPS-exposed RAW264.7 cells. In addition, the analysis of the mouse model of sepsis revealed that knockdown of CHI3L1 reduced the IL-1 β, IL-6, TNF-α, creatinine, blood urea nitrogen, alanine transaminase, and aspartate aminotransferase levels, and attenuated damage induced by sepsis to the heart, liver, lung, and kidneys in mice with sepsis. On the whole, the present study demonstrates that the lactylation modification of CHI3L1 promotes the development of sepsis. Thus, focusing on CHI3L1 expression changes may prove to be a promising approach for the treatment of sepsis in the future.
