单核细胞增生李斯特菌PrfA蛋白转录调控毒力基因表达的分子机制

单核细胞增生李斯特菌 (Listeria monocytogenes LM) 属于典型的细胞内寄生革兰氏阳性菌, 是WHO公布的四大食源性致病菌之一。LM不仅是人畜共患传染病李斯特菌病 (listeriosis) 的主要病原菌, 也是研究胞内感染和细胞介导的免疫应答的模式细菌。绝大多数LM毒力基因的转录表达受到PrfA蛋白的调控。本文简单介绍了LM侵染宿主细胞必需的毒力基因及其产物; 重点对毒力基因调节蛋白PrfA的结构和功能, PrfA调节毒力基因表达的主要方式最新进展进行了综述和讨论。     

单核细胞增生李斯特菌PrfA蛋白转录调控毒力基因表达的分子机制

单核细胞增生李斯特菌(Listeria monocytogenes LM)属于典型的细胞内寄生革兰氏阳性菌,是WHO公布的四大食源性致病菌之一.LM不仅是人畜共患传染病李斯特菌病(listeriosis)的主要病原菌,也是研究胞内感染和细胞介导的免疫应答的模式细菌.绝大多数LM毒力基因的转录表达受到PrfA蛋白的调控.本文简单介绍了LM侵染宿主细胞必需的毒力基因及其产物;重点对毒力基因调节蛋白PrfA的结构和功能,PrfA调节毒力基因表达的主要方式最新进展进行了综述和讨论.     

病毒感染相关基因微阵列的制备及其在HBV感染应答基因筛选中的应用

为筛选乙型肝炎(乙肝)病毒(HBV)感染应答基因,探讨HBV感染分子机理,采用生物信息学分析、筛选宿主细胞中与乙肝病毒、丙型肝炎(丙肝)病毒、流行性感冒(流感)病毒等感染密切相关的基因,设计并合成寡核苷酸探针,制备了含231种病毒感染相关基因的寡核苷酸微阵列.利用此微阵列比较HepG2细胞、HepG2.2.15细胞之间的基因表达谱差异,筛选乙肝病毒感染候选应答基因,从分子水平对乙肝病毒感染作用机理进行初步研究.制备的病毒感染相关基因表达谱微阵列的监测结果显示,阳性对照和看家基因探针出现较强信号,空白点样液和阴性对照探针未出信号,大部分基因探针信号强度在可分析范围内,上矩阵和下矩阵反映的基因表达情况一致,证明微阵列的特异性、敏感性、重复性都较好.HepG2.2.15与HepG2细胞基因表达谱比较结果显示,28个宿主基因在HepG2.2.15细胞中高表达,包括ASGR1、AFP、Fibronectin、APOC等基因;4个基因低表达,包括RRM1、ICSBP等基因.初步筛选获得HBV感染候选应答基因.此结果表明,制备的微阵列敏感性、特异性、重复性好,可为研究病毒宿主相互作用关系提供技术平台,应用此微阵列筛选获得的HBV候选应答基因可为揭示HBV感染的分子致病机理提供新的信息,为抗HBV药物研究提供潜在的作用靶点.     

结核分枝杆菌感染诱导的人巨噬细胞细胞因子及其调控基因的差异表达谱

研究人巨噬细胞细胞因子在抗结核分枝杆菌感染免疫中的作用。利用表达谱芯片技术研究细菌感染后,宿主巨噬细胞基因表达情况,在全局表达谱分析基础上,重点分析细胞因子及相关基因的表达,并比较无毒株和临床分离有毒株在诱导细胞因子及其调控基因表达方面的差异。结果显示细菌感染影响较多细胞因子及其调控基因的表达,如IFN、TNF、TGF、IL系列及其受体、NFkappa B和TLR受体等;首次报道IL19参与了抗结核分枝杆菌感染免疫。被临床株感染影响的细胞因子较无毒株广泛和丰富。细胞因子及相关基因参与了宿主细胞对感染细菌的免疫应答,有关细胞因子及相关基因在抗结核免疫中的作用有待进一步研究。     

乙型肝炎肝硬化患者外周血单核细胞基因表达谱研究

目的：探索基因表达谱技术在肝硬化形成的分子生物学机制研究及其诊断方法研究的应用。方法：应用含8192个人体cDNA的微阵列芯片和来自外周血单核细胞的标记cDNA,分析慢性乙型肝炎、肝炎肝硬化各15例基因表达谱。通过CenePix4000B扫描芯片仪和ImaGene3．0软件分析Cy3、Cy5两种荧光信号的强度和比值。结果：在8192个基因中,2组中筛选出有差异的基因60个,占0．73％,其中主要是炎症、凋亡基因、细胞外基质蛋白基因、细胞生长调节基因,占71．6％。结论：基因表达谱技术可为乙型肝炎肝硬化形成的分子生物学机制及其诊断研究提供大量有益的生物学信息。     

乙型肝炎肝硬化患者外周血单核细胞基因表达谱研究

目的 :探索基因表达谱技术在肝硬化形成的分子生物学机制研究及其诊断方法研究的应用。方法 :应用含 81 92个人体cDNA的微阵列芯片和来自外周血单核细胞的标记cDNA ,分析慢性乙型肝炎、肝炎肝硬化各 1 5例基因表达谱。通过GenePix40 0 0B扫描芯片仪和ImaGene3.0软件分析Cy3、Cy5两种荧光信号的强度和比值。结果 :在 81 92个基因中 ,2组中筛选出有差异的基因 60个 ,占 0 .73% ,其中主要是炎症、凋亡基因、细胞外基质蛋白基因、细胞生长调节基因 ,占 71 .6%。结论 :基因表达谱技术可为乙型肝炎肝硬化形成的分子生物学机制及其诊断研究提供大量有益的生物学信息     

基因芯片在研究病毒感染宿主基因表达谱中的应用

人类基因组计划提供的海量基因信息以及功能基因组学理论和技术的出现,为研究病毒与宿主相互作用提供了难得的历史机遇。病毒作为一种严格的细胞内寄生物,感染宿主后会引起宿主细胞形态和功能的改变。这些变化主要由于病毒感染导致宿主细胞基因表达变化所引起。通过基因芯片技术研究病毒感染宿主的基因表达谱变化,可以发现病毒感染宿主细胞在分子水平上的应答,从而为病毒性疾病的预防、诊断和临床治疗提供新的策略。     

siRNA介导的PRR11表达抑制导致肺癌细胞系基因表达谱变化的分析

Proline rich 11(PRR11)是本课题组鉴定的一个新的肿瘤相关基因。为研究PRR11介导肺癌发生发展相关的分子机制,本研究分析了PRR11表达被抑制后人肺癌细胞系H1299的全基因组基因表达谱的变化。首先,采用siRNA抑制H1299细胞中PRR11的表达,提取总RNA,采用基因芯片分析全基因组基因表达谱的变化。然后,对呈现差异表达的基因进行GO和Pathway富集分析,并对部分重要的候选基因进行定量RT-PCR验证。基因芯片结果表明,采用siRNA有效抑制H1299细胞中PRR11表达后,共有550个基因的mRNA水平出现明显变化,其中139个基因表达上调,411个基因表达下调。生物信息学分析结果表明,上述差异表达的基因显著富集于细胞周期和MAPK通路。定量RT-PCR验证分析结果表明,PRR11表达抑制后确实可导致多个与细胞周期和肿瘤发生发展密切相关的基因(包括DHRS2、EPB41L3、CCNA1、MAP4K4、RRM1、NFIB)呈现显著的表达变化。这些结果提示,PRR11可能通过上述通路和/或基因的表达变化参与肺癌的发生发展过程。     

应用 PPIN分析肝移植临床耐受患者PBMC基因表达特点

应用生物信息学方法分析肝移植临床耐受患者PBMC基因表达特征,筛选临床耐受关键基因。从GEO数据库获取19个肝移植临床耐受病例及22个非临床耐受病例基因表达谱数据。应用DAVID网络软件进行差异基因功能注释与聚类分析;通过Cytoscape软件的MiMI插件构建蛋白质相互作用网络(PPIN)筛选肝移植临床耐受关键基因。差异基因涉及蛋白质及RNA代谢、免疫应答、膜结构调节等复杂生物过程。PPIN网络分析获得10个临床耐受核心基因。我们的研究表明:肝移植临床耐受涉及外周血免疫细胞复杂的基因表达调控机制及蛋白质间相互作用;RNA的转录后加工及蛋白质降解在免疫耐受的形成中发挥了重要作用;RBM8A、DHX9、CBL、IKBKB、CSNK2A1、HSPA8等核心基因发挥重要的免疫调节功能。     

关于青岛文昌鱼SoxB2和SoxC基因的研究：进化保守性分析

大多数Sox基因在细胞命运决定和分化过程中起着重要的作用.本研究分离了青岛文昌鱼（Branchiostoma belcheri）的SoxB2和SoxC基因,对其预测的蛋白序列进行了序列比对、进化树分析以及基因时空表达分析.结果显示,文昌鱼SoxB2和SoxC虽然在进化树中不属于脊椎动物SoxB2和SoxC进化支,但它们在进化过程中基因的结构和表达部位保守性很高,都具有保守的HMG结构域;这两个基因在胚胎发育早期都在神经外胚层和原肠腔壁表达,而在成体中不仅存在于神经索中,还存在于肠、肝盲囊、鳃和卵母细胞中.SoxB2和SoxC基因表达部位的相似说明两者可能在中枢神经系统、性腺及免疫系统的胚胎发育和成体的免疫应答过程中共同起作用;两者与脊椎动物同源基因的表达部位相同说明这两个基因在进化过程中可能功能保守.     

Identifying significant genes and functionally enriched pathways in familial hypercholesterolemia using integrated gene co-expression network analysis

Familial hypercholesterolemia (FH) is a monogenic lipid disorder which promotes atherosclerosis and cardiovascular diseases. Owing to the lack of sufficient published information, this study aims to identify the potential genetic biomarkers for FH by studying the global gene expression profile of blood cells. The microarray expression data of FH patients and controls was analyzed by different computational biology methods like differential expression analysis, protein network mapping, hub gene identification, functional enrichment of biological pathways, and immune cell restriction analysis. Our results showed the dysregulated expression of 115 genes connected to lipid homeostasis, immune responses, cell adhesion molecules, canonical Wnt signaling, mucin type O-glycan biosynthesis pathways in FH patients. The findings from expanded protein interaction network construction with known FH genes and subsequent Gene Ontology (GO) annotations have also supported the above findings, in addition to identifying the involvement of dysregulated thyroid hormone and ErbB signaling pathways in FH patients. The genes like CSNK1A1, JAK3, PLCG2, RALA, and ZEB2 were found to be enriched under all GO annotation categories. The subsequent phenotype ontology results have revealed JAK3I, PLCG2, and ZEB2 as key hub genes contributing to the inflammation underlying cardiovascular and immune response related phenotypes. Immune cell restriction findings show that above three genes are highly expressed by T-follicular helper CD4⁺ T cells, naïve B cells, and monocytes, respectively. These findings not only provide a theoretical basis to understand the role of immune dysregulations underlying the atherosclerosis among FH patients but may also pave the way to develop genomic medicine for cardiovascular diseases.     

Cutting edge: molecular portrait of human autoimmune disease

Autoimmune diseases affect 3-5% of the population, are mediated by the immune response to self-Ags, and are characterized by the site of tissue destruction. We compared expression levels of >4,000 genes in PBMC of control individuals before and after immunization to those of individuals with four distinct autoimmune diseases. The gene expression profile of the normal immune response exhibits coordinate changes in expression of genes with related functions over time. In contrast, each individual from all autoimmune diseases displays a similar gene expression profile unrelated to the pattern of the immunized group. To our surprise, genes with a distinct expression pattern in autoimmunity are not necessarily "immune response" genes, but are genes that encode proteins involved in apoptosis, cell cycle progression, cell differentiation, and cell migration.     

Single-cell RNA sequencing reveals transcriptional profiles of monocytes in HBV-infected pregnant women during mid-pregnancy

There is a growing body of evidence that innate immunity also plays an important role in the progression of hepatitis B virus (HBV) infection. However, there is less study on systematically elucidating the characteristics of innate immunity in HBV-infected pregnant women. We compared the features of peripheral blood mononuclear cells in three healthy pregnant women and three HBV-infected pregnant women by single-cell RNA sequencing. 10 DEGs were detected between groups and monocytes were the main expression source of most of the DEGs, which involved in the inflammatory response, apoptosis and immune regulation. Meanwhile, qPCR and ELISA were performed to verify above genes. Monocytes displayed immune response defect, reflecting poor ability of response to IFN. In addition, eight clusters were identified in monocytes. We identified molecular drivers in monocytes subpopulations.TNFSF10+ monocytes, MT1G+ monocytes and TUBB1+ monocytes were featured with different gene expression pattern and biological function.TNFSF10+ monocytes and MT1G+ monocytes were characterized by high levels of inflammation response.TNFSF10+ monocytes, MT1G+ monocytes and TUBB1+ monocytes showed decreased response to IFN. Our results dissects alterations in monocytes related to the immune response of HBV-infected pregnant women and provides a rich resource for fully understanding immunopathogenesis and developing effective preventing HBV intrauterine infection strategies.     

Avian Resistance to Campylobacter jejuni Colonization Is Associated with an Intestinal Immunogene Expression Signature Identified by mRNA Sequencing

Campylobacter jejuni is the most common cause of human bacterial gastroenteritis and is associated with several post-infectious manifestations, including onset of the autoimmune neuropathy Guillain-Barré syndrome, causing significant morbidity and mortality. Poorly-cooked chicken meat is the most frequent source of infection as C. jejuni colonizes the avian intestine in a commensal relationship. However, not all chickens are equally colonized and resistance seems to be genetically determined. We hypothesize that differences in immune response may contribute to variation in colonization levels between susceptible and resistant birds. Using high-throughput sequencing in an avian infection model, we investigate gene expression associated with resistance or susceptibility to colonization of the gastrointestinal tract with C. jejuni and find that gut related immune mechanisms are critical for regulating colonization. Amongst a single population of 300 4-week old chickens, there was clear segregation in levels of C. jejuni colonization 48 hours post-exposure. RNAseq analysis of caecal tissue from 14 C. jejuni-susceptible and 14 C. jejuni-resistant birds generated over 363 million short mRNA sequences which were investigated to identify 219 differentially expressed genes. Significantly higher expression of genes involved in the innate immune response, cytokine signaling, B cell and T cell activation and immunoglobulin production, as well as the renin-angiotensin system was observed in resistant birds, suggesting an early active immune response to C. jejuni. Lower expression of these genes in colonized birds suggests suppression or inhibition of a clearing immune response thus facilitating commensal colonization and generating vectors for zoonotic transmission. This study describes biological processes regulating C. jejuni colonization of the avian intestine and gives insight into the differential immune mechanisms incited in response to commensal bacteria in general within vertebrate populations. The results reported here illustrate how an exaggerated immune response may be elicited in a subset of the population, which alters host-microbe interactions and inhibits the commensal state, therefore having wider relevance with regard to inflammatory and autoimmune disease.     

Gene expression profiling of hybridoma cells after bursal-derived bioactive factor BP5 treatment

Bursa of Fabricius is the acknowledged vital humoral immune system for B cell differentiation and antibody production. To study the molecular mechanism underlying the effect of bursal-derived BP5, we used gene microarray to analyze the genomic expression profiling of BP5-treated hybridoma cells. BP5 exhibited an immunomodulatory effect on antibody production in hybridoma cells and induced alterations in the gene expression profiles related to the immune-related biological processes, such as T cell activation and proliferation, B cell activation, B cell-mediated immunity, and cytokines cytokine production involved in immune response. In addition, 26 biological pathways associated with immunomodulatory functions were regulated in BP5-treated hybridoma cells, in which p53 signal pathway played an important role in antitumor. Among these regulated genes, 12 differentially expressed genes were verified by qRT-PCR. The activation of p53 activity by BP5 was further confirmed by p53 luciferase reporter assay and p53 expression. Our data revealed that bursal-derived BP5 could regulate various immune-related cellular processes, including antitumor factor p53 signal pathway, perhaps partially accounting for the reported immunomodulatory roles and novel antiproliferation on tumor cells functions of bursal-derived bioactive factor BP5.     

The Effect of Citalopram on Gene Expression Profile of Alzheimer Lymphocytes

Antidepressants are widely used in the treatment of mood disorders associated with dementia, however little information is available on their effect at the molecular level. In certain neurodegenerative disorders, such as in Alzheimer's disease, lymphocytes have been used to assess mirror changes that thought to occur in the brain. Gene expression profiles of lymphocytes from Alzheimer patients have been shown to differ from that seen with controls. To address this issue in light of antidepressant treatment, we used lymphocytes derived from Alzheimer's disease patients and control individuals to assess the impact of the selective serotonine reuptake inhibitor citalopram on gene expression using a cDNA microarray representing 3200 distinct human genes. Sequences that are differentially regulated after treatment with citalopram were identified and categorized based on similarities in biological functions. This analysis revealed that the overexpression of genes in control and Alzheimer white blood cells by citalopram are implicated in cell survival. Apart from this, citalopram did not markedly alter genes involved in other molecular functions in control cells. In contrast, alteration of genes implicated in ionic currents, cell-adhesion, immune mechanism, and adrenergic functions, were also observed in Alzheimer lymphocytes. The expression of genes of Alzheimer lymphocytes by citalopram is modulated differently which may correlate with the pathology.     

Identification of immune-associated signatures and potential therapeutic targets for pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) comprises a heterogeneous group of diseases with diverse aetiologies. It is characterized by increased pulmonary arterial pressure and right ventricular (RV) failure without specific drugs for treatment. Emerging evidence suggests that inflammation and autoimmune disorders are common features across all PAH phenotypes. This provides a novel idea to explore the characteristics of immunological disorders in PAH and identify immune-related genes or biomarkers for specific anti-remodelling regimens. In this study, we integrated three gene expression profiles and performed Gene Ontology (GO) and KEGG pathway analysis. CIBERSORT was utilized to estimate the abundance of tissue-infiltrating immune cells in PAH. The PPI network and machine learning were constructed to identify immune-related hub genes and then evaluate the relationship between hub genes and differential immune cells using ImmucellAI. Additionally, we implemented molecular docking to screen potential small-molecule compounds based on the obtained genes. Our findings demonstrated the density and distribution of infiltrating CD4 T cells in PAH and identified four immune-related genes (ROCK2, ATHL1, HSP90AA1 and ACTR2) as potential targets. We also listed 20 promising molecules, including TDI01953, pemetrexed acid and radotinib, for PAH treatment. These results provide a promising avenue for further research into immunological disorders in PAH and potential novel therapeutic targets.