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关键词： IL-23p19单克隆抗体   MRL/lpr狼疮小鼠   免疫表型   狼疮肾炎  

摘要： 目的 利用MRL/lpr狼疮样小鼠模型，观察白细胞介素23p19亚基（IL-23p19）单克隆抗体的治疗作用。方法 选用8周的雌性MRL/lpr小鼠共36只，随机分为6组。分别是PBS组（空白对照）、IgG组（同型IgG）、地塞米松（DEX）组（阳性对照）、IL-23p19单克隆抗体共分为3个浓度梯度治疗组：LD（1 mg/kg低浓度）、MD（3 mg/kg中浓度）、HD（10 mg/kg高浓度）治疗组。从小鼠12周龄时开始药物干预，尾静脉注射给药。尿蛋白检测试纸检测小鼠尿蛋白含量；ELISA检测小鼠血清中抗dsDNA抗体的含量；全自动生化分析检测仪检测小鼠血清肌酐及尿素氮含量；HE染色法及PAS染色分析肾脏组织病理变化；免疫荧光分析肾脏组织内IgG及C3免疫复合物的沉积情况；流式细胞术检测小鼠脾脏Th1、Th2、Th17、Tfh细胞和调节性T细胞（Treg）亚群表达；实时定量PCR检测脾脏相关转录因子表达情况。结果IL-23p19单克隆抗体可以减少MRL/lpr小鼠的尿蛋白水平，减少脾脏的肿大，改善肾功能和降低抗dsDNA抗体水平；缓解MRL/lpr小鼠的肾小球肾炎，并降低肾脏免疫复合物沉积；IL-23p19单克隆抗体可显著抑制MRL/lpr小鼠脾脏Th1、Th17细胞比例，上调Treg比例；同时显著下调脾脏转录因子T-bet（T-bet）和视黄酸受体相关孤儿受体γt（RORγt）mRNA水平，上调叉头框P3（FOXP3）mRNA水平。结论IL-23p19单克隆抗体对MRL/lpr小鼠具有显著的治疗效果，可能通过调控Th17/Treg之间细胞的平衡来发挥作用。

关键词： LAG3   HIV   NK   IFN-gamma   glycolysis   STAT1  

摘要： Natural killer (NK) cells are integral to the innate immune system and crucial for antiviral defense. NK cell activation and functional state are suppressed by inhibitory receptors. Lymphocyte activation gene 3 (LAG3) is an important inhibitory receptor, but the associated signaling pathways that regulate lymphocyte function remain to be elucidated. In addition, the effect of LAG3 on NK cell function during HIV infection and its specific mechanisms are unclear. In this study, we observed that LAG3 expression by NK cells is elevated in HIV-infected individuals and inversely correlated with CD4/CD8 ratio and CD4+ T cell count. LAG3+ NK cells produce lower levels of interferon-gamma (IFN-gamma), but LAG3-Fc protein significantly enhances NK cell function. The activation of LAG3 significantly inhibits IFN-gamma production and Ki67 expression by NK cells. Our transcriptome sequencing and in vitro data show for the first time that LAG3 not only regulates the transcription of MYC and several glycolysis-related enzyme genes via the PI3K/AKT/mTOR signaling pathway to inhibit glycolysis in NK cells but also suppresses the STAT1/IFNG pathway by upregulating PSAT1 expression, thus limiting IFN-gamma production by NK cells via these two different pathways. Overall, these results provide new insights and identify potential targets for immunotherapy of HIV *** demonstrate that lymphocyte activation gene 3 (LAG3) expression is upregulated on natural killer (NK) cells during HIV infection. LAG3 inhibits glycolysis in NK cells and also upregulates PSAT1 expression to suppress activation of the STAT1/IFNG pathway, thus restricting interferon-gamma production by NK cells. These results provide new clues to study the effects of LAG3 on the metabolism and functional exhaustion of NK cells and offer a potential target for the treatment of HIV.

关键词： 刚地弓形虫   表面抗原相关序列蛋白   SRS67   SRS20A   生物信息学分析   克隆  

摘要： 目的 采用生物信息学方法预测刚地弓形虫表面抗原相关序列蛋白SRS67和SRS20A的结构和免疫原性，并构建原核表达载体进行蛋白表达，以探讨重组蛋白功能及成为弓形虫病疫苗候选分子的潜力。方法 自ToxoDB数据库下载刚地弓形虫SRS67和SRS20A基因及氨基酸序列，于ORF Finder网站对SRS67和SRS20A基因开放阅读框进行分析。采用ProtParam软件预测刚地弓形虫SRS67和SRS20A蛋白相对分子质量、等电点、氨基酸组成及脂溶性指数等理化性质，采用ProtScale工具预测蛋白亲/疏水性，采用TMHMM软件预测蛋白跨膜区域，在SignalP-4.1网站预测蛋白信号肽，通过NPS@SPOMA和SWISS-MODEL网站预测蛋白的二级结构和三级结构，采用NetPhos-3.1程序预测蛋白磷酸化位点，应用Immuonmedicine Group程序预测蛋白抗原决定簇，通过IEDB网站和SYFPEITHI网站分别预测蛋白B细胞表位、辅助性T细胞（helper T cell,Th）抗原表位和细胞毒性T淋巴细胞（cytotoxic T lymphocyte,CTL）抗原表位，并采用VaxiJen 2.0软件对表位进行抗原性评分以筛选优势表位。根据ToxoDB数据库中刚地弓形虫SRS67和SRS20A基因序列合成引物序列，以弓形虫cDNA为模板，通过PCR反应扩增SRS67和SRS20A基因，并将扩增产物基因进行双酶切，回收目的片段。通过T4连接酶将目的片段转化至DH5α感受态细胞中，挑取阳性单菌落培养后提取pET-32a-SRS67和pET-32a-SRS20A重组质粒，并将其转化至表达感受态中进行小量诱导表达。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfatepolyacrylamide gel electrophoresis,SDS-PAGE）及Western blotting检测重组蛋白表达水平。结果 经生物信息学预测发现，刚地弓形虫SRS67基因与SRS20A基因长度分别为633、987 bp，分别具有7个和15个开放阅读框、编码210个和328个氨基酸。SRS67蛋白相对分子质量为23 135.65，有信号肽（D=0.590），无跨膜区，含有22个磷酸化位点、8个抗原决定簇，为亲水性蛋白；SRS20A蛋白相对分子质量为34 944.91，有信号肽（D=0.697）和跨膜区，含39个磷酸化位点、15个抗原决定簇，为亲水性蛋白。SRS67蛋白和SRS20A蛋白二级结构相似，均包含α-螺旋、β-折叠及无规则卷曲；三级结构模型呈典型球状特征，全球模型质量评估（Global Model Quality Estimation）分值分别为0.74和0.77。SRS67蛋白和SRS20A蛋白分别具有22个和39个磷酸化位点。SRS67蛋白平均抗原倾向指数为1.046 4，含有8个抗原决定簇；SRS20A蛋白平均抗原倾向指数为1.037 4，含有15个抗原决定簇。SRS67蛋白具有7个优势B细胞表位；SRS20A蛋白具有8个优势B细胞表位。对于Th细胞表位，SRS67与SRS20A分别筛选出10个和20个优势表位；对于CTL细胞表位，SRS67与SRS20A分别筛选出2个和3个优势表位。PCR扩增SRS67基因及SRS20A基因产物长度分别约为633 bp和987 bp，与预期相符。分别构建含目的基因的重组质粒，诱导表达发现SRS67重组蛋白在0.1 mmol/L IPTG诱导16 h后在沉淀中表达量最多、SRS20A重组蛋白在0.5 mmol/L IPTG诱导16 h后在沉淀中表达量最多，且SDS-PAGE及Western blotting检测结果表明重组蛋白表达成功。结论 SRS67和SRS20A蛋白具有多个细胞表位和良好免疫原性。本研究成功表达了重组SRS67和SRS20A蛋白，为后续研究蛋白功能及筛选有效的弓形虫病疫苗抗原提供了理论依据。

关键词： Prdm1   IFN-gamma   antisense oligonucleotides   intestinal epithelium   neonates   LncRNAs   Swi/Snf   Pias1   innate defense   Cryptosporidium  

摘要： Intestinal epithelial cells (IECs) serve as the front line of host defense in the intestine, with IFN signaling playing a critical role in regulating epithelial cell-intrinsic defense against intracellular pathogens. However, IFN-gamma-mediated antimicrobial defense is usually reduced in the gastrointestinal tract in infants, and the underlying mechanisms remain unclear. We previously observed that the lncRNA XR_001779380 promotes IFN-gamma-stimulated gene transcription in murine IECs. Interestingly, its interaction with Prdm1, a DNA-binding protein expressed in the neonatal but not adult intestinal epithelium, attenuates IFN-gamma-stimulated gene transcription, thereby contributing to suppression of IFN-gamma-mediated, epithelial cell-intrinsic defense in the neonatal intestine. In this study, we further investigated the role of Prdm1 in suppressing IFN-gamma response in murine neonatal IECs. Additionally, we explored the development of specific antisense oligonucleotides to interfere with XR_001779380-Prdm1 interaction to promote IFN-gamma response in IECs. Our data show that Prdm1 suppresses IFN-gamma-mediated gene transcription, and its induction inhibits IFN-gamma-stimulated cell-intrinsic defense against Cryptosporidium, an apicomplexan parasite and a leading cause of infectious diarrhea and diarrheal-related death in young children worldwide. Furthermore, antisense oligonucleotides designed to block Prdm1-XR_001779380 interaction can promote the IFN-gamma response in murine neonatal IECs, leading to enhanced cell-intrinsic anti-Cryptosporidium *** with adults, the innate antimicrobial defense of intestinal epithelium in neonates and infants is typically reduced, leading to increased susceptibility to infection;however, the underlying mechanisms remain incompletely understood. Cryptosporidium is a leading cause of infectious diarrhea and diarrheal-related death in children worldwide. Prdm1 is a DNA-binding protein that is expressed in ne

关键词： aCSF   artificial cerebrospinal fluid   Arc   activity-regulated cytoskeletal gene   BDNF   brain-derived neurotrophic factor   Bdnf   brain-derived neurotrophic factor gene   BLA   basolateral amygdala   C57BL/6NHsd   a strain of black mice used for stress testing   CA   1   Cornu Ammonis 1   CC   cage control   CD1   Hsd:ICR retired breeder mice used as aggressors   CD11b   Intergrin alpha M   CeA   central amygdala   cEPo   carbamoylated erythropoietin   CR   conditioned response   CS   conditioned stimulus   Creb1   cAMP responsive element binding protein 1 gene   CS   cEPo-treated Stay mice   ΔCt   difference between target gene value of corresponding endogenous reference gene   ΔΔCt   fold change of target gene expression relative to reference sample, normalized to reference gene   CR   cued response   DG   Dentate Gyrus   EC. Escape cEPo-treated   Egr1   early growth response gene 1   EPo   erythropoietin   EpoR   erythropoietin receptor   Epor   erythropoietin receptor gene   EV   Escape Vehicle-treated   g   gram(s)   ga   gauge   Gapdh   housekeeping gene   GFAP,   glial fibrillary acidic protein gene   Hcrtr1   Orexin receptor 1 gene   Hcrtr2   Orexin receptor 2 gene   icv   intracerebroventricular   iDG   single intra-dentate gyrus   NIH   National Institutes of Health   NIMH   National Institute of Mental Health   PBS   phosphate buffer solution   PTSD   post-traumatic stress disorder   s   seconds   SAM   Stress Alternatives Model   SC   Stay cEPO-treated   SIP   Social Interaction Preference   SV   Stay Vehicle-treated   SVZ   subventricular zone   TNF   α   tumor necrosis factor alpha   US   unconditioned stimulus  

摘要： The hormone and trophic factor Erythropoietin (EPo) promotes red blood cell production and has neurotrophic effects, modulating behavior and promoting neural plasticity, like neurogenesis. Modifying EPo by attaching a carbamoyl group (cEPo) results in similar neuronal effects without erythropoietic actions. We hypothesize that neuroplastic and learning effects of cEPo may be dependent on its action in dorsal dentate gyrus of the hippocampus, where neurogenesis occurs. The Stress Alternatives Model (SAM) is a 4-day social stress and decision-making paradigm with a large novel aggressor that provides opportunities to avoid interaction via escape routes. Early, male test mice display stable Escape (avoiding aggression) or Stay (acquiescence to aggressor) behavioral phenotypes. In these studies, mice were given a single intracerebroventricular (icv; 100 ng), or single intra-dentate gyrus (iDG; 10 ng) injection of cEPo or vehicle. By Day 4, 30% of icv cEPo treated mice and 37.5% of iDG cEPo treated mice reversed their phenotype (Stay to Escape). Mice receiving vehicle injections did not change. Normalization of social preference, and reduction in fear freezing behavior, after cEPo treatment, coincide with transcriptional changes of orexin receptors (Hcrtr & Hcrtr) in the hippocampus, including phenotype- and treatment-dependent alterations in learning-associated molecular signaling molecules. Hippocampal Hcrtr moderately colocalize with EPo receptors (Epor) in the dentate gyrus, while Hcrtr is expressed with Epor in CA. Anxiolytic actions of cEPo during social interaction indicate mechanisms that influence learning in stressful situations, suggesting that cEPo may be a novel agent for treatment of comorbid conditions related to anxiety, depression, and PTSD.

关键词： ADH   alcohol dehydrogenase   ALDH2   aldehyde dehydrogenase 2   ASCA   anti-   Saccharomyces cerevisiae   -IgG antibodies   Clec   C-type lectin domain containing   CYP2E1   cytochrome P450 2E1   IL   interleukin   IgA   immunoglobin A   ILC3   innate lymphoid cells type 3   IFNγ   interferon-gamma   iNKT cells   invariant natural killer T cells   LPS   lipopolysaccharide   FMT   fecal microbiota transplantation   FXR   farnesoid X receptor   MAIT cells   mucosa-associated invariant T cells   MAMPs   microbial-associated molecular patterns   MELD   model of end-stage liver disease   Reg3   regenerating islet-derived protein 3   TNF   tumor necrosis factor   TNFR1   TNF receptor 1   TLR   Toll-like receptor  

摘要： Alcohol-associated organ damage is a major cause of morbidity and mortality worldwide, with the liver being the primarily affected organ. Emerging evidence highlights the gut microbiota as a key driver in alcohol-associated liver disease. Changes in the microbiota composition, microbial translocation, and a dysregulated immune response culminate in inflammation and tissue injury. Persistent liver injury eventually promotes disease progression from steatohepatitis to fibrosis and cirrhosis. In this review, we provide an overview of microbiota alterations observed in patients with chronic alcohol consumption and explore the mechanisms by which microbiota contributes to alcohol-associated damage across various organ systems. Emphasis is placed on changes in the gut microbiota and its role in alcohol-associated liver disease, the disease condition with the most robust evidence linking microbiota-related changes to organ injury. Additionally, we highlight key areas where further research is needed to address unresolved questions. Finally, we discuss emerging therapeutic strategies targeting the microbiota to treat alcohol use disorders and prevent alcohol-associated liver diseases.

关键词： Bleomycin induced Scleroderma   Chronic inflammation   Dihydromyricetin   Oxazolone induced dermatitis   RORγt   T cells  

摘要： Systemic sclerosis is a chronic inflammatory disease affecting the connective tissue of the human body by inducing fibrosis mainly in skin and lungs. Atopic dermatitis is another chronic inflammatory skin diseases that affects one-fifth of the population in developed countries. Dysregulation of the immune cells are one of the major characteristic features behind establishment of these two diseases. Several medicines are used to treat these diseases, but they have several side effects and high production cost. Low side effect and easy availability has generated renewed interest in studying plant derived medicines. Dihydromyricetin (Ampelopsin), a flavonoid compound extracted from stem and leaves of Ampelopsis grossedentata has shown potent anti-inflammatory property in vitro. In this study, bleomycin induced scleroderma and oxazolone induced dermatitis model were established in male BALB/c mice to check the in vivo efficacy of dihydromyricetin. Immunophenotyping and cytokine production were investigated by flow cytometry; immunofluorescence of skin was studied using confocal microscopy. We observed oral application of dihydromyricetin significantly reduced the inflammatory parameters in both diseases. We also found that dihydromyricetin dose dependently reduced the percentage of IL17A producing T cell population and reduced the total expression of RORγt in diseased T cells. Furthermore, we also found stable and strong binding of dihydromyricetin- RORγt protein complex. Stability of protein-ligand complex was examined by MD simulation study. We suggest that dihydromyricetin alleviates scleroderma and dermatitis symptoms by regulating the RORγt pathway.

关键词： Black carp   IFN   IRF3   STAT2   Ubiquitination  

摘要： Interferon (IFN) regulatory factor 3 (IRF3) is a critical transcription factor involved in inducing IFN production. However, the regulatory mechanisms of piscine IRF3 remain inadequately explored. Here, we report that the signal transducer and activator of transcription 2 (STAT2) of black carp (Mylopharyngodon piceus) functions as a positive regulator in the black carp IRF3 (bcIRF3)-mediated IFN signaling pathway. Knockdown of bcSTAT2 in vivo facilitates viral replications, and the mRNA levels of bcSTAT2 gene increase after IFN stimulation in host cells. Additionally, overexpression of bcSTAT2 promotes the activation of interferon stimulated response element (ISRE) ex vivo. Co-immunoprecipitation assays identify the interaction between bcSTAT2 and bcIRF3, and both molecules exhibit similar subcellular distributions in immunofluorescent staining assay. When co-expressed, bcSTAT2 improves the protein level of bcIRF3, and enhances the bcIRF3-mediated IFN production and antiviral capabilities. Mechanistically, we demonstrate that bcSTAT2 significantly enhances the nuclear translocation of bcIRF3. Besides, bcSTAT2 enhances K29-linked ubiquitination, while reduces K33- and K48-linked ubiquitination of bcIRF3. Taken together, our data concludes that bcSTAT2 positively regulates bcIRF3-mediated antiviral activity by regulating its ubiquitination and facilitating its nuclear translocation, which has shed a light on the regulation of IFN signaling.

关键词： 银屑病   炎症小体   白细胞介素-18   炎症小体抑制剂  

摘要： 白细胞介素-18（interleukin-18，IL-18）具体多种生物学功能的细胞因子，参与炎症、肿瘤和自身免疫性疾病的发生发展过程。近年来，银屑病临床和动物研究发现，炎症小体（inflammasome）的活化参与了银屑病的发病机制，激活的炎症小体进一步介导IL-18的成熟与释放。银屑病患者及其动物模型的皮损和血清中IL-18的表达水平显著增高，且与银屑病严重程度呈正相关。因此，靶向炎症小体激活以及介导IL-18的释放可能成为银屑病的潜在治疗方向。本文将围绕炎症小体激活介导的IL-18释放及炎症小体抑制剂在银屑病中的作用进行综述。