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关键词： 骨髓来源的巨噬细胞(BMDM)   锌指蛋白Krüppel样转录因子9(KLF9)   1型单纯疱疹病毒(HSV-1)   机制  

摘要： 目的 明确锌指蛋白Krüppel样转录因子9（KLF9）在1型单纯疱疹病毒（HSV-1）诱导巨噬细胞I型干扰素表达中的作用及机制。方法 通过琼脂糖凝胶电泳、实时荧光定量PCR及Western blot法检测KLF9在Klf9-/-（gKO）小鼠骨髓来源的巨噬细胞（BMDM）中的敲除效果；RNA-seq 筛选gKO小鼠及对照小鼠Klf9+/+（WT）BMDM中HSV-1刺激后Klf9下游关键差异基因；实时荧光定量PCR验证HSV-1刺激后干扰素β1（Ifnb1）及下游干扰素刺激基因（ISG）如干扰素诱导蛋白与四肽重复 1（Ifit1）、干扰素刺激的核酸外切酶基因 20（Isg20）、胆固醇25-羟化酶（Ch25h）及2'-5'-寡聚腺苷酸合成酶（Oasl1）的表达情况；ELISA检测HSV-1刺激后gKO及WT小鼠BMDM上清中β干扰素（IFN-β）的含量；Western blot法检测HSV-1刺激后gKO及WT小鼠BMDM中磷酸化干扰素调节因子 3（p-IRF3）、干扰素调节因子 3（IRF3），p-IRF7、IRF7，磷酸化的核因子κB p65（p-NF-κB p65）、核因子κB p65（NF-κB p65）的表达；CUT-Tag及ChIP-qPCR技术检测KLF9在Ifnb1上的结合。结果 KLF9在gKO小鼠的BMDM中表达明显减少；RNA-seq 结果显示BMDM中Klf9缺失导致I型干扰素信号通路受损；实时荧光定量PCR表明Klf9敲除后BMDM中除了Isg20以外，Ifit1、Ch25h及Oasl1表达均明显降低；ELISA显示Klf9敲除导致BMDM分泌的IFN-β明显减少。在机制研究方面，Western blot显示Klf9的缺失不影响p-IRF3、p-IRF7及p-NF-κB p65的表达；CUT-Tag及ChIP-qPCR显示KLF9能直接结合至Ifnb1的启动子区促进其转录。结论 KLF9对巨噬细胞抵抗HSV-1感染至关重要。

关键词： LCN-2   TNF-alpha   Porcine intramuscular adipocyte   Inflammatory response  

摘要： Lipocalin-2 (LCN-2) has a variety of biological functions and produces various effects on adipocytes, such as promoting cell apoptosis, inhibiting preadipocyte differentiation, and weakening insulin signaling. Tumor necrosis factor-alpha (TNF-alpha) is one of the first secreted products discovered in adipocytes, which plays an important role in the regulation of adipose metabolism. This experiment was conducted to investigate the regulatory effect of LCN-2 on TNF-alpha induced inflammatory response in porcine intramuscular adipocytes. Porcine intramuscular adipocytes were cultured in vitro and treated with LCN-2 overexpression or silencing plasmids. After TNF-alpha treatment, the expressions of LCN-2, interleukin 6 (IL-6), IL-8, and IL-1 beta were detected by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Western blot analysis was used to detect the expression of phosphatidylinositol 3-kinase (PI3K), total protein kinase B (Akt), and phosphorylated Akt (pAkt) proteins. After the action of the PI3K/Akt pathway inhibitor LY294002, the effects of LCN-2 overexpression on IL-6, IL-8, and IL-1 beta were evaluated. The results showed that TNF-alpha induced LCN-2 expression in a dose-/time-dependent manner. Overexpression or silencing of LCN-2 had an impact on TNF-alpha induced IL-6, IL-8, and IL-1 beta in porcine intramuscular adipocytes. Overexpression of LCN-2 significantly promoted inflammatory factors IL-6, IL-8, and IL-1 beta secretion while silencing of LCN-2 inhibited the secretion of these inflammatory factors (P < 0.01). Overexpression of LCN-2 significantly increased the expression of pAkt protein in cells, while silencing of LCN-2 decreased pAkt protein expression (P < 0.01). After blocking the PI3K/Akt signaling pathway, compared with the control group, overexpression of LCN-2 affected IL-6, IL-8, and IL-1 beta secretion, but the impact was not significant (P > 0.05). This study su

关键词： T cell receptor   TCR-T therapy   PRAME   phase I clinical trial   hematological malignancies   AML   MDS   MM  

摘要： Background/objectives: MDG1011 is a Preferentially Expressed Antigen in Melanoma (PRAME)-specific autologous T cell receptor (TCR) T cell therapy for HLA-A*02:01-positive patients. Data from the first-in-human (FIH) clinical trial, CD-TCR-001, are reported here regarding treatment feasibility, safety, tolerability, and clinical activity of MDG1011 in patients with relapsed/refractory (r/r) acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), and multiple myeloma (MM). Methods: Nine of thirteen enrolled patients received MDG1011 at dose levels ranging from 0.1 to 5 x 106 TCR-T cells per kg body weight. In addition to clinical assessments, immune monitoring of cytokines associated with cytokine release syndrome (CRS), presence and persistence of MDG1011, and changes in levels of PRAME mRNA were used to assess safety and potential biological activity at defined time points. Results: The treatment was well tolerated. No dose-limiting toxicities (DLTs) were observed, and the most common serious adverse events were associated with lymphodepleting chemotherapy and/or disease progression. Various parameters, such as measurable clinical responses in two patients, the occurrence of CRS in two additional patients, and reductions in PRAME mRNA levels in bone marrow (BM) or peripheral blood (PB) in seven patients, served as signs of the clinical and biological activity of MDG1011 TCR-T therapy. Conclusions: Patients enrolled in the phase 1 part of CD-TCR-001 displayed signs of potential clinical and biological activity of MDG1011 among the small number of patients studied. Advanced disease stage and rapid progression in the r/r AML patients limited clinical impact. The acceptable safety profile of MDG1011 merits further investigation of this TCR-T therapy, potentially in patients at an earlier stage of their disease and with lower tumor burden.

关键词： Immune Checkpoint Inhibitors   Immune related adverse event - irAE   Major histocompatibility complex - MHC   Diabetes  

摘要： Immune checkpoint inhibitors (ICIs) have improved survival of patients with cancer, yet they pose risks of immune-related adverse events (irAEs). ICI-induced insulin-dependent diabetes mellitus (ICI-DM) is a life-threatening and life-altering irAE. Previously, we reported a high incidence of a germline missense variant in NLRC5, a key class I transcription activator, among patients with ICI-DM compared with similarly treated patients who did not develop ICI-DM. Our purpose was to validate this finding in additional ICI-treated patients and study effects of the NLRC5 variant on expression of class I major histocompatibility complex (MHC) antigen presentation genes. We assessed the prevalence of the C>T missense variant at chr16:57 025 515 (NLRC5Pro191Leu) in germline DNA from an additional 33 patients with ICI-DM and in patients with ICI-induced colitis (n=15), ICI-induced hypothyroidism (n=19) and ICI-induced hypophysitis (n=17). The 1,000 Genomes Project was used for comparison. We assessed peripheral blood mononuclear cells from 16 individuals with or without the NLRC5 variant, studying expression of NLRC5 and select downstream target genes, before and after stimulation with interferon-gamma. We validated the higher prevalence of NLRC5Pro191Leu in a non-overlapping cohort of patients with ICI-DM compared with the general population (51.5% vs 12.8%, p<0.0001). The prevalence of NLRC5Pro191Leu in ICI-induced colitis or thyroiditis patients did not significantly differ from the general population, while the prevalence in ICI-induced hypophysitis was somewhat higher (21.6%, p=0.048). We found greater increases in messenger RNA expression of NLRC5 (p=0.007), TAP1 (p=0.0002), B2M (p=0.0005), HLA-G (p=0.04), PSMB8 (p=0.03) and PSMB9 (p=0.01) in NLRC5Pro191Leu cells stimulated with interferon-gamma compared with NLRC5(WT) cells. A similar trend was observed for HLA-A (p=0.09). We confirm the significantly higher prevalence of the NLRC5Pro191Leu variant in patients with IC

关键词： immune checkpoint inhibitor   NSCLC   melanoma   cytokines   Il-2   IL-10   TNF-alpha  

摘要： This prospective single-center study examined associations between serum cytokines-TNF-alpha, IL-2, and IL-10-and outcomes in stage IV non-small cell lung cancer (NSCLC, n = 43) and melanoma (n = 15) patients treated with Nivolumab at the Oncology Institute in Cluj-Napoca, Romania. Cytokines were measured at baseline (NSCLC: n = 43;melanoma: n = 15), 3 months (NSCLC: n = 20;melanoma: n = 7), and 6 months (NSCLC: n = 10;melanoma: n = 5). Melanoma patients showed sustained IL-2 and TNF-alpha increases, while NSCLC patients displayed heterogeneous cytokine dynamics. In NSCLC, elevated IL-10 at 3 months correlated with shorter survival (rho = -0.51, 95% CI -0.78 to -0.12, p = 0.022) and poorer response (rho = -0.65, 95% CI -0.86 to -0.23, p = 0.002). TNF-alpha showed a borderline association with response (rho = -0.44, 95% CI -0.74 to 0.01, p = 0.050). In melanoma, 3-month TNF-alpha was inversely associated with survival (rho = -0.82, 95% CI -0.97 to -0.15, p = 0.023) and response (rho = -0.90, 95% CI -0.99 to -0.39, p = 0.006). Strong inter-cytokine correlations were observed (NSCLC: TNF-alpha vs. IL-10, rho = 0.60, 95% CI 0.19-0.82;melanoma: rho = 0.93, 95% CI 0.44-0.99). Baseline cytokines had limited utility, particularly in melanoma due to the small sample size. The most informative finding was the association of elevated 3-month IL-10 with adverse outcomes in NSCLC. These results support the value of dynamic cytokine monitoring in immunotherapy and warrant validation in larger cohorts.

摘要： Inflammation promotes heart failure (HF) development, and inhibition of IL-12β simultaneously attenuates interleukin-12 (IL-12) and interleukin-23 (IL-23), two important proinflammatory cytokines. In this study, we used IL-12β knockout (KO) mice to test the hypothesis that genetic inhibition of IL-12β would attenuate transverse aortic constriction (TAC)-induced cardiac inflammation, hypertrophy, and dysfunction, as well as the consequent lung remodeling. IL-12β KO in male and female mice significantly attenuated TAC-induced cardiac dysfunction as evidenced by improved left ventricular (LV) ejection fraction and fractional shortening. IL-12β KO also significantly ameliorated the TAC-induced increase of LV weight, left atrial weight, lung weight, right ventricular (RV) weight, and their ratios to body weight or tibial length in male and female mice. In addition, IL-12β KO significantly attenuated TAC-induced LV leukocyte infiltration, cardiomyocyte hypertrophy, fibrosis, and the consequent lung inflammation and remodeling. Moreover, IL-12β KO reduced TAC-induced alterations of LV gene profile associated with inflammation and fibrosis, as shown by bulk LV RNA sequencing. Furthermore, we found that IL-12β KO significantly attenuated TAC-induced LV accumulation of multiple immune cell subsets, activation of CD4 and CD8 T cells, and the percentage of central memory CD4 and CD8 T cells in the cardiac drainage lymph nodes. Finally, IL-12β KO mice showed significantly reduced IFNγ CD8 and CXCR3 CD8 T cells in the drainage lymph nodes as compared with WT after TAC. These findings collectively demonstrate that IL-12β plays a critical role in systolic overload-induced LV inflammation, remodeling, and dysfunction, likely through cardiac immune cell infiltration.

摘要： Not available.

关键词： compressive force   IL-6   inflammatory response   melatonin   periodontal ligament cells   root resorption   YAP  

摘要： Background To explore the effects and mechanisms of melatonin on orthodontically induced root resorption (OIIRR) by focusing its role on IL-6 secretion in human periodontal ligament cells (hPDLCs).Methods Cytokine array was used to identify the key inflammatory cytokine secreted by hPDLCs under excessive compressive force. Western blot analysis and immunohistochemistry staining (IHC) were conducted to examine the key proteins in the YAP/P65/IL-6 signaling pathway both in in vitro and in vivo experiments. Verteporfin, a YAP inhibitor, was used to confirm the involvement of p-YAP and its downstream signaling factor p-P65. Root resorption volume was assessed using micro-computed tomography (micro-CT).Results Cytokine array revealed that excessive compressive force significantly elevated interleukin-6 (IL-6) levels in hPDLCs. IHC indicated that both IL-6 and melatonin receptor 1 (MT1) were highly expressed in hPDLCs on the compressive side in the mouse OIIRR model. in vitro experiments demonstrated that the levels of p-YAP and p-P65 significantly increased when compressive force was applied to hPDLCs, and melatonin reversed this effect. Furthermore, Verteporfin produced effects similar to melatonin on IL-6 expression and the YAP/P65 signaling pathway. Micro-CT analysis showed noticeable root resorption in the mouse OIIRR model, which was significantly reduced following intraperitoneal injection of melatonin. IHC staining further confirmed that the YAP/P65/IL-6 signaling pathway was inhibited on the compressive side of the mouse OIIRR model after melatonin *** Melatonin was able to inhibit root resorption in the mouse OIIRR model and reduced IL-6 secretion in hPDLCs under compressive force by suppressing the YAP/P65 signaling *** Language Summary This study explored how melatonin, a natural hormone, protects against root resorption during orthodontic treatment. Specifically, we focused on how excessive compressive force induces IL-6 production

摘要： Interleukin-18 (IL-18) is an acute-phase proinflammatory molecule crucial for mediating viral clearance by activating T helper 1 CD4+ T cells, cytotoxic CD8+ T cells and natural killer (NK) cells. Here, we show that mature IL-18 is generated in the thymus following numerous distinct forms of tissue damage, all of which cause caspase-1-mediated immunogenic cell death. We report that IL-18-stimulated cytotoxic NK cells limit endogenous thymic regeneration, a critical process that ensures the restoration of immune competence after acute insults such as stress, infection, chemotherapy and radiation. NK cells suppress thymus recovery by aberrantly targeting thymic epithelial cells, which act as the master regulators of organ function and regeneration. Together, our data reveal a new pathway regulating tissue regeneration in the thymus and suggest IL-18 as a potential therapeutic target to boost thymic function. Moreover, given the enthusiasm for IL-18 as a cancer immunotherapy due to its capacity to elicit a type 1 immune response, these findings also offer insight into potential off-target effects.

关键词： Shigella   Shigellosis   intestinal epithelial cells   macrophages   IL-12   IFN-g   neutrophils   mouse model   innate immunity  

摘要： Bacteria of the genus Shigella replicate in intestinal epithelial cells and cause shigellosis, a severe diarrheal disease that resolves spontaneously in most healthy individuals. During shigellosis, neutrophils are abundantly recruited to the gut and have long been thought to be central to Shigella control and pathogenesis. However, how shigellosis resolves remains poorly understood due to the longstanding lack of a tractable and physiological animal model. Here, using our newly developed Nlrc4(-/-)Casp11(-/-) mouse model of shigellosis, we unexpectedly find no major role for neutrophils in limiting Shigella or in disease pathogenesis. Instead, we uncover an essential role for macrophages in the host control of Shigella. Macrophages respond to Shigella via Toll-like receptors (TLRs) to produce IL-12, which then induces IFN-gamma, a cytokine that is essential to control Shigella replication in intestinal epithelial cells. Collectively, our findings reshape our understanding of the innate immune response to Shigella.