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关键词： LRRK2   Crohn's Disease   Parkinson's Disease   B Cells   Interleukin-4  

摘要： Mutations in Leucine-Rich Repeat protein Kinase 2 (LRRK2) are associated with Parkinson's disease (PD) and Crohn's disease (CD), but the regulation of LRRK2 during inflammation remains relatively unexplored. Here we describe the development of a flow cytometry-based assay to assess LRRK2 activity in individual cells and the generation of an EGFP-Lrrk2 knock-in reporter mouse to analyse cell-specific LRRK2 expression. Using these tools, we measured LRRK2 levels and activity in murine splenic and intestinal immune cells and in human blood. Anti-CD3 induced inflammation increases LRRK2 expression and activity in B cells and monocytes, while in mature neutrophils, inflammation stimulates activity but reduces LRRK2 expression. A kinase-activating PD-associated LRRK2-R1441C mutation exacerbates inflammation-induced activation of LRRK2 specifically in monocytes and macrophages. We identify IL-4 as a novel T-cell-derived factor that upregulates LRRK2 expression and activity in B cells, replicating inflammatory effects observed in vivo. Our findings provide valuable new insights into the regulation of the LRRK2 pathway in immune cells, crucial for understanding LRRK2 and its therapeutic potential in inflammatory diseases such as CD.

关键词： Biomarker   Immunology   Interleukin   Tumor microenvironment   Urothelial cancer  

摘要： BACKGROUND:Interleukin-6 (IL-6) is associated with adverse clinical outcome in cancer patients. In bladder cancer (BC) patients, higher IL-6 serum levels have been linked with adverse pathologic features, worse overall survival (OS) and cancer-specific survival (CSS). IL-6 is being investigated as a therapeutic target. However, concentrations in tumor-tissue are not investigated in detail. Objective of this study is to analyze the prognostic value of IL-6 in BC patients and to investigate its concentration in tumor tissue. METHODS:In this single center prospective observational study, preoperative serum samples of 179 BC patients undergoing radical cystectomy were collected between September 2019 and September 2022. Tumor-tissue of additional 20 patients was collected during transurethral resection or radical cystectomy for investigation of IL-6 in tumor tissue supernatant. IL-6 concentration was measured by ELISA. RESULTS:Median serum IL-6 concentration was 5.4 pg/mL. High serum IL-6 was an independent predictor of OS (HR 1.95; 95% CI, 1.07-3.55; P = .03) and CSS (HR 2.31; 95% CI, 1.14-4.68; P = .02) in multivariate Cox regression analyses. Patients with advanced tumor stage, lymph node metastasis, and larger tumor size had significantly higher preoperative serum IL-6 concentration (all P < .01). In tumor tissue supernatant, IL-6 concentration was higher in muscle-invasive BC, with a median of 715.4 pg/mL, as opposed to 20.7 pg/mL in pTa tumor stage (P < .01). CONCLUSIONS:Serum IL-6 is a strong predictor of poor survival rates and adverse pathologic features in BC patients. IL-6 concentrations in tumor tissue supernatant correlate with tumor stage. The role of IL-6 in theranostics of bladder cancer deserves more attention.

关键词： IL-23   IL-23p19 subunit inhibitors   guselkumab   CD64   pain   immune-mediated inflammatory diseases  

摘要： Immune-mediated inflammatory diseases (IMIDs) are a heterogeneous group of disorders-including gastroenteric, rheumatologic, dermatologic, neurologic, and systemic autoimmune conditionscharacterized by chronic inflammation and intermittent flares. 1 While a wide range of proinflammatory mediators are involved in the pathophysiology of IMIDs, in certain subsets such as Crohn's disease, psoriasis, and psoriatic arthritis, specific cytokines including tumor necrosis factor (TNF) and interleukin-23 (IL-23) have emerged as key drivers of inflammation. 1 In these conditions, TNF and IL-23 can act as "master regulators" by orchestrating immune cell activation and sustaining inflammatory responses, thus representing important therapeutic targets. However, this centrality is not consistent across all IMIDs; in other diseases such as multiple sclerosis or systemic lupus erythematosus, these pathways may be less prominent or therapeutically unsuitable 1 .Over the years, IL-23, a member of the IL-12 cytokine family, has emerged as a hierarchically dominant regulatory cytokine capable of reprogramming immune cells to induce and maintain a highly proinflammatory state 2 . It acts as a "master switch" in the development and persistence of certain IMIDs. IL-23 profoundly affects both innate immune cells (e.g., macrophages, dendritic cells, ILC3s, and neutrophils) and adaptive immunity by promoting the differentiation of naïve T cells into the TH17 phenotype 3 . Notably, a subset of mononuclear phagocytes, such as macrophages, dendritic cells or CD64brightCD163 -CD14brightCD1c -CD1a -inflammatory monocyte-like cells, are key sources of IL-23 in response to various physiological and pathological stimuli 3 .Structurally, IL-23 is a heterodimeric cytokine composed of the proteins p19 and p40. While p19 is the unique subunit of IL-23, p40 is the common subunit shared with IL-12 3 . In recent years, the monoclonal antibodies risankizumab, guselkumab, mirikizumab, and tildrakizumab, which

关键词： skimmianine   breast tumor   PCNA   TNF-alpha   immunohistochemistry   bioinformatics  

摘要： Background/Objectives: Breast cancer continues to be a major global health challenge, driving the urgent need for innovative therapeutic strategies. This study evaluates the anticancer and immunomodulatory potential of skimmianine in breast cancer through a comprehensive approach, integrating biochemical, histopathological, immunohistochemical, and bioinformatics analyses. Methods: Thirty-six female Wistar albino rats were divided into three groups: control, 7,12-dimethylbenz[a]anthracene (DMBA)-induced breast cancer, and DMBA + skimmianine (n = 12/group). Breast cancer was induced with a single oral dose of 50 mg/kg DMBA in sesame oil. After 16 weeks, skimmianine (40 mg/kg) was administered intraperitoneally for four weeks. Serum CA15-3 levels were measured via enzyme-linked immunosorbent assay (ELISA). Histopathological assessment was performed using hematoxylin and eosin (H&E) staining, and proliferating cell nuclear antigen (PCNA) and tumor necrosis factor-alpha (TNF-alpha) were evaluated immunohistochemically. Pathway and hub gene analyses were performed using Cytoscape, functional annotation with Enrichr, and immune analyses via the Tumor and Immune System Interaction Database (TISIDB) and Sangerbox. Results: The tumor burden in the animals increased after DMBA induction compared to the control groups (0.00 +/- 0.00% vs. 89.00 +/- 6.60%, respectively, p < 0.001), while skimmianine treatment significantly reduced the tumor burden in the animals (49.00 +/- 9.40%, vs. DMBA group, p = 0.191). Histopathological analysis showed DMBA-induced structural disorganization and malignant clustering, whereas skimmianine preserved ductal structures and mitigated the damage. Compared to the control group, DMBA administration markedly elevated serum CA15-3 levels (0.23 +/- 0.06 ng/mL vs. 8.57 +/- 1.01 ng/mL, respectively), along with PCNA (13.0 +/- 3.0% vs. 25.0 +/- 4.0%, respectively) and TNF-alpha (8.4 +/- 1.7% vs. 34.0 +/- 5.3%, respectively) expression, indicating active t

关键词： Bispecific T cell engager - BiTE   Cytokine   Immune Checkpoint Inhibitor   Immunotherapy   Vaccine  

摘要： Background Immune checkpoint inhibitors and cytokines have revolutionized tumor treatment but are still limited by dose-dependent toxicity and efficacy. In situ vaccine platforms based on intelligent microbes are promising therapeutic strategies that sustainably deliver drugs locally without causing severe systemic *** In this study, we have innovatively engineered a non-pathogenic, adjuvant-acting Mycobacterium smegmatis (M. smegmatis) that co-expresses a programmed cell death-ligand 1 (PD-L1) inhibitor and an interleukin-15 (IL-15) cytokine complex containing the interleukin-15 receptor alpha (IL-15R alpha) sushi domain (Ms-PDL1scfv-IL15).Results We demonstrate that the fusion protein of PD-L1 inhibitor and IL-15 cytokine systemically binds mouse or human PD-L1 and maintains IL-15 stimulatory activity. The bifunctional Ms-PDL1scfv-IL15 overcomes resistance to PD-L1 blockade, recruits numerous immune cells in situ, induces dendritic cells (DCs) maturation, initiates the M1 antitumor polarization of macrophages, increases the proliferation and activation of natural killer cells and tumor-infiltrating CD8+ T cells, inhibits regulatory T cells, elicits abscopal effects, stimulates rapid tumor regression, prevents metastasis, and leads to long-term survival in several syngeneic tumor mouse models. We also found that the combination of Ms-PDL1scfv-IL15 with granulocyte-macrophage colony-stimulating factor (GM-CSF) synergistically stunted the tumor progress and stasis. Moreover, intratumoral administration of Ms-PDL1scfv-IL15 can capture tumor antigen fragments, and boost DCs presentation of antigens, which remarkably initiates tumor antigen-specific immune response, leading to durable tumor regression and specific antitumor *** In summary, the engineered M. smegmatis can recruit and activate innate and adaptive antitumor immune responses, offering a potent cancer immunotherapy strategy to treat patients with cold tumors or resistance to checkpo

关键词： Human DRG   Multi-omics   Proteomics   Sexual dimorphism  

摘要： The peripheral nervous system (PNS) plays a critical role in pathological conditions, including chronic pain disorders, that manifest differently in men and women. To investigate this sexual dimorphism at the molecular level, we integrated quantitative proteomic profiling of human dorsal root ganglia (hDRG) and peripheral nerve tissue into the expanding omics framework of the PNS. Using data-independent acquisition (DIA) mass spectrometry, we characterized a comprehensive proteomic profile, validating tissue-specific differences between the hDRG and peripheral nerve. Through multi-omic analyses and in vitro functional assays, we identified sex-specific molecular differences, with TNFα signalling emerging as a key sexually dimorphic pathway with higher prominence in men. Genetic evidence from genome-wide association studies further supports the functional relevance of TNFα signalling in the periphery, while clinical trial data and meta-analyses indicate a sex-dependent response to TNFα inhibitors. Collectively, these findings underscore a functionally sexual dimorphism in the PNS, with direct implications for sensory and pain-related clinical translation.

关键词： HSV-1   ocular   herpes   HLA transgenic rabbit   CD8(+) T cell   prime/pull/keep vaccine  

摘要： Herpes simplex virus type 1 (HSV-1) continues to be one of the most prevalent viral infections globally, with approximately 3.72 billion individuals affected worldwide. A clinical herpes vaccine is still lacking. In the present study, a novel prime/pull/keep vaccine was tested in a human leukocyte antigen transgenic rabbit model of ocular herpes (HLA-A*0201 Tg rabbit). Ten asymptomatic (ASYMP) CD8(+) T-cell peptide epitopes and 3 CD4(+) T-cell epitopes were selected from the HSV-1 glycoproteins D and B (gD and gB), viral tegument proteins (VP11/12 and VP13/14), and the DNA replication-binding helicase (UL9), all preferentially recognized by CD8(+) and CD4(+) T cells from "naturally protected" HSV-1-seropositive healthy ASYMP individuals (who never had recurrent corneal herpetic disease). HLA Tg rabbits were ocularly infected with HSV-1, then during latency at day 30 post-infection, the rabbits were ocularly vaccinated with a recombinant neurotropic AAV8 vector (10(7)GC/ eye) encoding for the 10 CD8(+) T-cell peptide and 4 CD4(+) T-cell peptide (prime), T-cell attracting CXCL-11 (pull), and T-cell keeping IL-2/IL-15 cytokines (keep). The rabbits were followed up for corneal disease and viral loads in tears for 28 days. The frequency, function, and protective efficacy of HSV-specific CD8(+) T cells induced by the prime/pull/keep vaccine were assessed in the trigeminal ganglia (TG), cornea, spleen, and peripheral blood. Compared to the mock group (unvaccinated), the peptides/CXCL11/IL-2/IL-15 vaccine generated frequent resident CD8(+) T cells that infiltrated the TG. In ocularly HSV-1-infected and prime/pull/keep vaccinated rabbits, CD8(+) T cell mobilization and retention into TG were associated with a significant reduction in corneal herpes infection and disease. These findings draw attention to the novel prime/pull/keep therapeutic vaccine strategy to mobilize and retain antiviral T cells to tissues protecting them against herpetic infection and disease.

关键词： stratified cardiac rehabilitation   elderly   coronary heart disease   CRP   exercise tolerance  

摘要： Coronary heart disease (CHD) is a highly prevalent disease in the elderly population, with atherosclerosis as its pathology, which can also be viewed as a chronic inflammatory response of the organism. Regular moderate-intensity exercise can direct the immune response toward an anti-inflammatory state, which is beneficial for improving the health and exercise tolerance. In cardiac rehabilitation, attention to the management of inflammatory factors as well as the improvement of exercise endurance is beneficial for the rehabilitation of elderly patients with coronary artery disease. This study investigates the impact of tiered cardiac rehabilitation programs on levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-alpha), and the capacity for physical exertion in older CHD patients. From March 2020 to April 2022, 94 elderly patients with CHD visiting our institution were recruited and randomly allocated into either a control group or an observation group, each comprising 47 participants. The standard care group participated in traditional rehabilitation exercises, whereas the experimental group received customized, tiered, cardiac rehabilitation interventions. We assessed the variations in CRP and tumor necrosis factor alpha (TNF-alpha) levels, along with exercise capacity, before and after treatment in both groups. The result shows that significant reductions in CRP and TNF-alpha levels were seen in the experimental group after 4 and 12 weeks, compared to the standard care group. Analysis showed clear trends in CRP and TNF-alpha changes over the interventions, with the experimental group showing better results. CRP levels decreased consistently, while TNF-alpha levels stayed stable. The experimental group also showed improvements in physical endurance measures compared to the control group. Interleukin 6 (IL-6) and fibrinogen (Fib) in the observation group decreased compared with the control group (P < 0.01). After 12 weeks of treatment, CRP and TNF-al

关键词： Japanese encephalitis virus (JEV)   non-structural protein 1 (NS1)   DDX3X   IFN-beta   ISGs  

摘要： Japanese encephalitis virus (JEV) is the causative agent of Japanese encephalitis, which poses great threats to the pig farming industry and human health. To establish infection, JEV has evolved sophisticated strategies to overcome the innate immune responses and finish its life cycle. Previous studies have shown that non-structural protein 1 (NS1) is closely related to its pathogenesis, while its molecular mechanism remains elusive. In this study, host protein ATP-dependent RNA helicase DEAD-box helicase 3 X (DDX3X) was screened to bind with NS1, and both ectopically expressed and virally encoded NS1 further confirmed their interaction. We also proved that the beta-roll and wing subdomains of NS1 were responsible for their interaction. In DDX3X-overexpressing cells, the replication of JEV was markedly inhibited, while the viral titers were elevated in DDX3X-silencing cells, indicating that DDX3X might serve as an anti-viral factor during JEV infection. Mechanically, overexpression of DDX3X promotes interferon-beta (IFN-beta) transcription, while its transcription was decreased in DDX3X-silencing cells. Consistent with IFN-beta, some interferon-stimulated genes (ISGs), including protein kinase R (PKR), myxovirus resistance 1 (MX1), guanylate-binding protein 1 (GBP1), and bone marrow stromal antigen 2 (BST2), were also positively related to the DDX3X expression. Taken together, JEV NS1 blocks IFN-beta production by interacting with DDX3X to evade the host's innate immune response and facilitate virus replication. This finding will deepen our understanding of JEV immune-evasion strategies and provide targets for JEV *** study focused on JEV, a threat to pig farming and human health. The key finding is that NS1 binds to host protein DDX3X via its beta-roll and wing subdomains. JEV NS1 evades the host immune response by interacting with DDX3X to restrain type I interferon production. These results deepen our understanding of JEV's immune-evasion s

关键词： endothelial cells   HUVECs   IL-1   IL-33   IL-36  

摘要： Background Blockade of IL1RAP (interleukin 1 receptor associated protein) was recently shown to reduce atherosclerosis in mice, but the effect on human vascular cells is largely unknown. Targeting the IL1RAP coreceptor represents a novel strategy to block the IL1RAP-dependent cytokines IL (interleukin)-1, IL-33, and IL-36. In the present study, we aimed to evaluate the role of novel antibodies targeting IL1RAP to reduce the effects of IL-1 beta, IL-33, or IL-36 gamma in human vascular *** Expression of IL1RAP was observed in human atherosclerotic plaques by immunohistochemistry and microarray and in endothelial cells by flow cytometry. Endothelial cells were cultured with IL-1 beta, IL-33, or IL-36 gamma cytokines with or without IL1RAP antibodies and analyzed with Olink proteomics, ELISA, Western blot, and real-time quantitative polymerase chain reaction. The functional effect of IL1RAP antibodies on endothelial cells were analyzed with adhesion and permeability *** Olink proteomics showed inhibition of the inflammatory proteins LIF (leukemia inhibitory factor), OPG (osteoprotegerin), CCL4 (C-C motif chemokine ligand 4), and MCP-3 (monocyte chemoattractant protein 3) by IL1RAP-blockade in endothelial cells after IL-1 beta stimulation. In addition, the IL1RAP antibodies inhibited IL-1 beta, and IL-33 induced IL-6 and IL-8 secretion. Secretion of MCP-1 (monocyte chemoattractant protein 1) was induced by IL-1 beta, IL-33, and IL-36 gamma, and subsequently was inhibited by IL1RAP antibodies. Similar effects were found on mRNA expression level. Endothelial expression of the adhesion markers ICAM1, VCAM1, and SELE were significantly reduced by IL1RAP antibodies, and neutrophil adhesion to endothelial cells induced by IL-1 beta and IL-33 was reduced by IL1RAP blockade. In human atherosclerotic lesions, IL1RAP expression correlated with markers of inflammation like IL6, IL8, and *** IL1RAP-targeting antibodies can reduce the expression of