TLR4, IL10RA, and NOD2 mutation in paediatric Crohn’s disease patients: an association with Mycobacterium avium subspecies paratuberculosis and TLR4 and IL10RA expression

Mycobacterium avium subspecies paratuberculosis (MAP) has been implicated in the pathogenesis of Crohn’s disease (CD). The role of CD susceptibility genes in association with these microbes is not known. Sixty-two early onset paediatric CD patients and 46 controls with known MAP status were analysed for an association with 34 single nucleotide polymorphisms (SNPs) from 18 CD susceptibility genes. Functional studies on peripheral blood mononuclear cells (PBMCs) were conducted on 17 CD patients with known CD mutations to assess IL-6, IL-10, and TNF-α expression upon stimulation with MAP precipitated protein derivative (PPD) and lipopolysaccharide (LPS). In addition, surface expression of IL10R and TLR4 on resting B cells, NK cells, T cells, and monocytes was assessed. A mutation in TLR4 (rs4986790) and IL10RA (rs22291130) was significantly associated with MAP-positive CD patients compared to MAP-negative CD patients (27.6 vs. 6.1 %, p = 0.021, and 62.1 vs. 33.3 %, p = 0.024, respectively). PPD and LPS significantly increased IL-6, IL-10, and TNF-α production in PBMCs. IL-10 and TNF-α production were significantly lower in a subgroup of CD patients (5/12) with a known NOD2 mutation. Receptor for IL-10 was significantly higher expressed on NK cells (CD56low) and on NK T cells harbouring a NOD2 mutations compared to wildtype cells (p = 0.031 and 0.005, respectively). TLR4 was significantly higher expressed on NK cells (CD56high) harbouring a NOD2 mutations compared to wildtype cells (p = 0.038).     

Association study between the IL4, IL13, IRF1 and UGRP1 genes in chromosomal 5q31 region and Chinese Graves’ disease

Graves disease (GD) was believed to be a polygenic disease. Several chromosomal regions were linked to GD, and the 5q31 chromosome regions containing several interleukin genes cluster were worth observing. In this study, IL4, IL13, IRF1 and UGRP1 genes were sequenced, and 5, 3, 7 and 7 polymorphisms respectively were discovered. Then an extended association study for the attracting polymorphisms was performed with 146 sporadic Graves patients, 142 unrelated controls and the 54 multiplex Graves families. However, the genotype and allele frequency distribution of these polymorphisms had similar distribution between the Graves patients and unrelated controls, and transmission disequilibrium tests indicated that none of them showed dominant transmission from heterozygous parents to the affected offsprings. Comparison of the clinical variables of the Graves patients indicated that the onset ages of the patients carrying TT at IRF1 6477 T/G locus were younger than those having variant allele (TG, GG); the difference was of statistical significance (P=0.005, Pc=0.020). Our association study revealed that, IL4, IL13, IRF1 and UGRP1 genes in chromosomal 5q31 regions might not confer susceptibility to Chinese GD. But those individuals who were TT homozygous at IRF1 6477 T/G locus seemed to be attacked by GD much earlier than others.Huang Wei and Teng Weiping equally contributed to this work     

PSS病人血清IL—2,IL—4,IL—6水平的测定

本文用生物测定法技术检测了３０例进行性系统性硬化症患者血清中白细胞介素２、白细胞介素４、白细胞介素６的水平，结果表明：在ＰＳＳ患者血清中，ＩＬ－２、ＩＬ－４的检出率显著高于对照组，ＩＬ－６的检出率虽然高于对照组，但是在统计学上无显著差异。结果提示：ＩＬ－２、ＩＬ－４可能在该病的发病机制中起一定作用。     

TLR2和TLR4的TLR/IL-1 receptor结构与功能

Toll样受体(Toll like receptor,TLR)是架接固有免疫和适应性免疫的桥梁。迄今为止,已鉴定的人TLR有10种,分别命名为TLR1~TLR10,TLR是一种Ⅰ型跨膜蛋白,由胞外富含亮氨酸重复序列(LRR)结构域,胞内保守的Toll/IL-1受体(TIR)结构域和跨膜结构域构成。TLRs在识别病原体相关分子模式后,其信号的特异转导主要取决于TLRs的TIR功能域与下游接头分子的TIR功能域的特异相互作用。然而,TLR2和TLR4的TIR功能域在其与下游接头分子Mal和MyD88相互作用、以及TLRs同源或异源二聚化方面的作用模式存在明显的差异。本文就TLR2和TLR4的TIR结构与功能进行简要综述,有助于我们进一步了解TLR TIR功能域与下游接头分子的相互作用。     

CpG-ODN-mediated TLR9 innate immune signalling and calcium dyshomeostasis converge on the NFκB inhibitory protein IκBβ to drive IL1α and IL1β expression

Sterile inflammation contributes to many pathological states associated with mitochondrial injury. Mitochondrial injury disrupts calcium homeostasis and results in the release of CpG-rich mitochondrial DNA. The role of CpG-stimulated TLR9 innate immune signalling and sterile inflammation is well studied; however, how calcium dyshomeostasis affects this signalling is unknown. Therefore, we interrogated the relationship beτween intracellular calcium and CpG-induced TLR9 signalling in murine macrophages. We found that CpG-ODN-induced NFκB-dependent IL1α and IL1β expression was significantly attenuated by both calcium chelation and calcineurin inhibition, a finding mediated by inhibition of degradation of the NFκB inhibitory protein IκBβ. In contrast, calcium ionophore exposure increased CpG-induced IκBβ degradation and IL1α and IL1β expression. These results demonstrate that through its effect on IκBβ degradation, increased intracellular Ca²⁺ drives a pro-inflammatory TLR9-mediated innate immune response. These results have implications for the study of innate immune signalling downstream of mitochondrial stress and injury.     

IL—1,IL—6与肿瘤的关系

ＩＬ－１和ＩＬ－６在肿瘤发生发展中具双重性作用，其即可抑制某些肿瘤的发生，亦可促进一些肿瘤的增殖、转移。其抗肿瘤活性或促瘤生长均有靶细胞或靶组织特异性，在不同肿瘤中有不同作用。     

IL—1Ra,IL—1Ra基因多态性与溃疡性结肠炎

IL-1Ra,IL-1Ra基因多态性与溃疡性结肠炎（UC）的相关性研究是从遗传基因角度探讨了UC的发病，通过对该方面深入研究可望进一步认识到UC发病机制，为其诊断及治疗开辟新途径。     

β-Glucan attenuates TLR2- and TLR4-mediated cytokine production by microglia

Microglia, the resident immune cells of the brain, are activated in response to any kind of CNS injury, and their activation is critical for maintaining homeostasis within the CNS. However, during inflammatory conditions, sustained microglial activation results in damage to surrounding neuronal cells. β-Glucans are widely recognized immunomodulators, but the molecular mechanisms underlying their immunomodulatory actions have not been fully explored. We previously reported that β-glucans activate microglia through Dectin-1 without inducing significant amount of cytokines and chemokines. Here, we show that particulate β-glucans attenuate cytokine production in response to TLR stimulation; this inhibitory activity of β-glucan is mediated by Dectin-1 and does not require particle internalization. At the molecular level, β-glucan suppressed TLR-mediated NF-κB activation, which may be responsible for the diminished capacity of microglia to produce cytokines in response to TLR stimulation. Overall, these results suggest that β-glucans may be used to prevent or treat excessive microglial activation during chronic inflammatory conditions.     

重型病毒性肝炎患者PBMCs中TNF—α,IL—1α和IL—4…

本实验应用半定量ＲＴ－ＰＣＲ检测了８例急重肝，１３例亚急重肝和１６例慢重肝患者ＰＢＭＣｓＴＮＦ－α，ＩＬ－１α和ＩＬ－４ｍＲＮＡ的表达水平对其意义进行了探讨。结果表明，急重肝和亚急重肝患者ＰＢＭＣｓＴＮＦ－α和ＩＬ－αｍＲＮＡ的表达水平较正常人明显增高，且与内毒素血症和病情轻重相关，而慢重肝仅ＴＮＦ－αｍＲＮＡ略增高，提示ＴＮＦ－α和ＩＬ－１α在急重肝和亚急重肝的发病机理中可能起重要的作用，而在慢     

Atorvastatin protects rat brains against permanent focal ischemia and downregulates HMGB1, HMGB1 receptors (RAGE and TLR4), NF-κB expression

Inflammatory processes play a key, mainly detrimental role in the pathophysiology of ischemic stroke. Currently, HMGB1-induced NF-κB activation pathway has been recognized as a key contributor to the proinflammatory response. It has been proved that chronic administration and pre-treatment with statins could protect brain tissue against ischemic injury. However, little is known about the effects of statins in the acute phase after cerebral ischemia. Thus, this study investigated the atorvastatin's protective role and the underlying mechanisms in cerebral ischemia. After middle cerebral artery occlusion (MCAO), atorvastatin was administered immediately. We found that atorvastatin dramatically improved neurological deficits, reduced brain water contents and infarct sizes at 24 h after stroke. Moreover, the over-expression of HMGB1, RAGE, TLR4 and NF-κB induced by ischemia was significantly attenuated by atorvastatin.     

TLR4 and TLR2 activation is differentially associated with age during Parkinson’s disease

Parkinson’s disease (PD) is an age-related neurodegenerative disease characterized by loss of dopaminergic neurons associated with neuroinflammation. Toll-like receptors (TLRs) are expressed in peripheral blood leukocytes and also in neurons and glial cells mediating inflammation. This study aimed to investigate the peripheral blood leukocyte response to TLR2 and TLR4 agonists in young and elderly PD patients. Two groups of patients with PD were evaluated (≤ 55 years old and ≥ 65 years old), age-matched with healthy controls (n = 26). Severity of PD was evaluated by Unified Parkinson’s Disease Rating Scale (UPDRS). Whole blood cultures were stimulated with lipopolysaccharide (LPS), a TLR4 agonist or Pam₃Cys (Pam), a TLR2 agonist. Tumor necrosis factor alpha (TNFα) and interleukin 10 (IL-10) were measured by immunoenzimatic assay. 6 h-TNFα production was increased after TLR4 stimulation, mainly in young PD patients, whereas TLR2-induced TNFα and IL-10 levels were decreased in PD patients independent of age (p < 0.05). A reverse correlation between LPS-induced TNFα production and age was observed in PD patients and controls, but TNFα induced by TLR2 agonist was not associated with age of PD patients or controls. TNFα production induced by TLR4 but not by TLR2 was reversely associated with the age at PD onset and disease duration. No associations between UPDRS scores and cytokine levels were detected. In conclusion, TLR4 and TLR2 responses seem to be differentially affected during PD. Data suggest that TLR2 deficiency in periphery is independent of age of the patients, age at PD onset, or PD duration.     

IL—2,IL—4基因转染后肿瘤细胞表面ICAM—1分子的表达及其作用

为了进一步研究细胞因子基因转染后肿瘤细胞体内致瘤原性、免疫原性变化的分子机制，将ＩＬ－２、ＩＬ－４基因转染入Ｂ１６黑色素瘤细胞，观察了其体内接种后致瘤原性变化，通过ＦＡＣＳ法检测了其细胞表面粘附分子－１（ＩＣＡＭ－１）的表达水平，分析了其细胞表面ＩＣＡＭ－１表达水平以及它们与肿瘤细胞对ＬＡＫ、ＣＴＬ等免疫效应细胞杀伤敏感性变化的关系。结果表明，ＩＬ－２、ＩＬ－４基因转染后Ｂ１６细胞体内致瘤原性明显     

HMGB1及TLR2、TLR4在类风湿关节炎中的表达及意义

 目的 探讨HMGB1及TLR2、TLR4在类风湿关节炎患者中的表达。方法 选取活动期类风湿关节炎患者29例,非活动期20例及对照者18例。用酶联免疫吸附试验（ELISA）检测血清HMGB1表达,用多色免疫荧光流式细胞术（FCM）检测外周血单核细胞表面TLR2和TLR4的表达率和蛋白相对含量。结果 活动期RA患者血清HMGB1含量和外周血CD14+单核细胞表面TLR2、TLR4的表达率及蛋白相对含量明显高于非活动期和对照者;非活动期RA患者TLR4显著高于对照组。活动期RA患者血清HMGB1与外周血CD14+单核细胞表面TLR2、TLR4相对蛋白含量表达呈显著正相关。结论 RA患者血清HMGB1及外周血单核细胞表面TLR2、TLR4的表达可能与RA病情发生发展有关,且具协同作用。     

Molecular Diagnosis of Severe Combined Immunodeficiency��Identification of IL2RG, JAK3, IL7R, DCLRE1C, RAG1, and RAG2 Mutations in a Cohort of Chinese and Southeast Asian Children

Severe combined immunodeficiencies (SCID) are a group of rare inherited disorders with profound defects in T cell and B cell immunity. From 2005 to 2010, our unit performed testing for IL2RG, JAK3, IL7R, RAG1, RAG2, DCLRE1C, LIG4, AK2, and ZAP70 mutations in 42 Chinese and Southeast Asian infants with SCID adopting a candidate gene approach, based on patient's gender, immune phenotype, and inheritance pattern. Mutations were identified in 26 patients, including IL2RG (n?=?19), IL7R (n?=?2), JAK3 (n?=?2), RAG1 (n?=?1), RAG2 (n?=?1), and DCLRE1C (n?=?1). Among 12 patients who underwent hematopoietic stem cell transplantation, eight patients survived. Complications and morbidities during transplant period were significant, especially disseminated bacillus Calmette-Guérin disease which was often difficult to control. This is the first cohort study on SCID in the Chinese and Southeast Asian population, based on a multi-centered collaborative research network. The foremost issue is service provision for early detection, diagnosis, management, and definitive treatment for patients with SCID. National management guidelines for SCID should be established, and research into an efficient platform for genetic diagnosis is needed.     

感染日本血吸虫病小鼠肝脏,脾脏和结肠IL—4,IL—5和I …

目的：为了解感染日本血吸虫后小鼠不同脏器的免疫反应，我们观察了小鼠肝，脾、小肠Ｔｈ２因子的变化并探讨其意义。方法：采用ＡＢＣ免疫组化法，并用多媒体病理图文定量分析，动态研究感染后８，１０，１２周ＩＬ－４，ＩＬ－５，ＩＬ－１０的变化。结果：在感染小鼠肝脏ＩＬ－４明显高于正常对照，并随感染时间的延长而逐步升高，ＩＬ－５和ＩＬ－１０亦有相同变化，但ＩＬ－４最高，ＩＬ－１０次之，ＩＬ－５居后；在感染小鼠的     

TLR4与中枢神经系统损伤

As a receptor mediating the transmembrane signal transduction in the innate immunity, Toll-like receptor 4 (TLR4) is a bridge between innate immunity and required immunity, and plays an important role when signal-transducing of some cells are activated. Recent reports show that TLR4 expresses in the different glial cells and strongly links to the innate immune activation and inflammatory response in the central nervous system (CNS). TLR4 plays a key role in the processes of brain damage by infection of the CNS, stroke, cerebral hemorrhage and trauma. In this review, we concentrate on recent findings regarding the progress of function and mechanism of TLR4 in the processes of the CNS damage in various diseases.