博士生课程细胞因子

8-30KDCK-CKRs,Cytockines are low-molecular-weight regulatory proteins or glycoproteins secreted by white blood cells and various cells (vascular endothelial cell, epidermic cell and fibroblast ) in body in response to a number of stimuli.,Cytokine,macrophage,Just as hormones serve as messenger of the endocrine system, so cytokine sever as messengers of the immune system. Unlike endocrine hormones , though, which exert their effects over large distances, cytokines general act locally.,Cytokines can exert different actions,IL-2Proliferation, survival,Example:,ChemokinesLeukocyte recruitment,Acute phase proteins, IL-6TNFIL-33,Properties of cytokines,细胞因子调节IgG类别转换,多效性,拮抗性,重叠性,协同性,Biological effects of the cytokines,Anti-infection and antitumorImmune regulationApoptosisStimulating the proliferation and differentiation of hemopoietic stem cellsPromote the growth and differentiation of various cellsParticipation and regulation of the inflammatory responsesCytokine abnormalities in the pathogenesis in certain diseases,Cytokines during hematopoiesis,interleukin, IL （白细胞介素）colony stimulating factor,CSF（集落刺激因子）interferon,IFN（干扰素）tumor necrosis factor,TNF （肿瘤坏死因子）Chemokine, CK （趋化因子）growth factor,GF （生长因子）,Cytokines,Cytokines of the innate and adaptive immune system,Cytokines of innate immune system,TNF, IL-1 and IL-6: pleiotropic mediators of inflammation (activators of endothelium, immune cells, liver and brain)Chemokines: regulators of immune cell adhesion at infection sitesIL-12: inducer of IFN-, increase of cytotoxic activityType I IFN: inducers of antiviral state,InterleukinInterleukins are secreted by some leukocytes and act upon other leukocytes, IL-1IL-35 have been identified.,Interleukin-1, IL-1,The release of TNF-a by macrophages induces local protective effects, but TNF-a can have damaging effects when released systemically (septic shock).,Tumor necrosis factor,TNF,The cytokines that induce directly apoptosis of tumor cells, including TNF, TNF (Lymphotoxin, LT） , FASL, CD70L, CD30L, CD40L, OX40L, TRAIL (TNF related apoptosis-inducing ligand),Chemokine,chemokines, a group of low-molecular-weight cytokines including interleukin 8, that affect chemotaxis and other aspects of leukocyte behavior.,1999年国际命名委员会根据CK的结构特征命名依据N端前两个半胱氨酸(Cys)的相对位置不同分为:,IL-12 produced by macrophages and dendritic cells is a key cytokine to fight intracellular microbes (bacteria and viruses)IL-12 induces IFN-, an activator of macrophages (killing of phagocytosed microbes) and Th1 cell differentiationIL-12 also increases cytotoxic activity of CD8 and NK cells,IL-12,The cytokines that resist viral infection, and interfere viral replication, including IFN, IFN, IFN, IFN, IFN ( type I ); IFN ( type II ),type I Interferon,Antiviral effects of Type I Interferons (IFN-a/b),Integrative signaling of cytokines during innate immune responses,Cytokines of adaptive immune system,IL-2: prototypic lymphocyte growth factorIL-4: inducer of Th2 cells, B cell differentitation factorIFN-: inducer of Th1 cells,IL-2 (又称T细胞生长因子,TCGF) ： 细胞来源：主要由T细胞产生。 作用方式：以自分泌和旁分泌方式发挥效应。 主要生物学功能： （1）活化T细胞，促进细胞因子产生； （2）刺激NK细胞增殖，增强NK杀伤活性及产生 细胞因子；诱导LAK细胞产生 （3）促进B细胞增殖和分泌抗体； （4）激活巨噬细胞。,IL-4 induces Isotype switching against extracellular pathogens, such as helminths and athropodes,IL-4,Pleiotropic activities of IFN-,IFN- is secreted by activated T cells after IL-12 stimulation during antimicrobial responses (intracellular bacteria and viruses)Th1 cells help B cell sotype switch to IgG for complement binding and Fc-binding on phagocytes strong macrophage activator,Anti-inflammatory cytokines,IL-10IL-10: counteracts IFN- by blocking IL-12 production by macrophages, and expression of costimulators and MHC IIIL-10-deficient mice develop inflammatory bowel disease Tr1TGF-TGF- is secreted by regulatory T cells (Th3) to decrease immune responsesThe TGF receptor is a dimer of ALK and TGF-RII which signal through Smad transcription factorsAbsence of TGF or TGF-RII induces autoimmunity in mice,Additional key cytokines,IL-5: activator of eosinophils, also inducing production of additional eosinophils from bone marrow progenitorsIL-13: similar to IL-4, but in addition stimulates mucus production and tissue fibrosis in allergic diseases GM-CSF: promotes maturation of bone marrow cells to monocytes and dendritic cellsG-CSF: generated at site of infection but acting as endocrine hormone to mobilize neutrophils during inflammation (clinical use),Colony stimulating factor, CSF,The cytokines that stimulate hemopoietic cells to form cell colony, Participate hemopoietic function are called CSF, for example GM-CSF,G-CSF,M-CSF, Epo,Tpo,是一类可介导不同类型细胞生长和分化的细胞因子。根据其功能和作用的靶细胞不同，分别命名为：表皮生长因子（epidermal growth factor, EGF)血管内皮生长因子（vascular endothelial growth factor, VEGF)成纤维细胞生长因子（fibroblast growth factor, FGF)神经生长因子(nerve growth factor, NGF)血小板衍生的生长因子（platelet derived growth factor, PDGF) 肝细胞生长因子（hepatic growth factor, HGF),Growth factor,VEGF Receptor Signaling,Growth factors applied to angiogenesis,VEGF promotes Tumor Angiogenesis,TH1 and TH2 subsets secrete different cytokines,TH1- cell-mediated functions; inflammationTH2- humoral immunity,TH1 and TH2 subsets regulate each other,CD4+ T cells isolation and differentiationSpleens were mechanically dissociated, followed by depletion of erythrocytes. Naive CD4+ T cells were isolated by negative selection. CD4+ T cells were cultured in medium supplemented with anti-mouse CD3 and anti-mouse CD28. rmIL-2 plus anti-mouse IL-4 and anti-mouse IFN-antibodies (Th0 conditions) or anti-mouse IL-4 plus rmIL-2 and rmIL-12 (Th1 conditions) or rmIL-2, rmIL-4 plus anti-mouse IFN- antibody (Th2 conditions) were added.,After 4 days, T cells were washed and reseeded on anti-mouse CD3-coated plates in the absence or in the presence of anti-mouse CD28 for 24 hours, and cytokine secretion was determined by ELISA.,The red line represents GATA3 staining, the black line corresponds to isotype control.,细胞因子受体分类,主要包括免疫球蛋白受体超家族(1gRSF)、I型细胞因子受体家族、型细胞因子受体家族(干扰素受体家族)、型细胞因子受体家族(肿瘤坏死因子受体家族，TNFRF)和七次跨膜受体家族(又称G蛋白偶联受体家族),Cytokine receptors belong to families of receptor proteins, each with a distinctive structure,The various members of the interleukin-1 (IL-1) family and their receptors,趋化因子受体（ GPCRs，G-protein coupled receptors ） 属于7次跨膜受体超家族，胞内环状结构与Gi/Go蛋白偶联,Signaling of cytokine receptors,Type I/II cytokine receptors: Jak-STAT pathwayTNF receptors: TRAF (TNF-RII, CD40) or by death domains (TNF-RI, Fas)TIR-domain: Toll-like receptor signaling through IRAKReceptor-associated kinases: TGF- receptors, c-kitG-protein-coupled signaling,Class I and Class II cytokine receptors signal by a rapid pathway using receptor-associated kinases to activate specific transcription factors.,Jak-STAT signaling pathways, common to many cytokine receptors,4 Jak proteins known: Jak1-3 and Tyk26 STATs known: STAT1-4, 5a, 5b and 6Receptor dimerization central for signalingCombination of Jak/STATs confers specificity,JAK/STATs Used by Key Cytokines,IL-5-Eosinophil,Cytokine imbalance during inflammation,CXCR4（T细胞）与CCR5（巨噬细胞） 是HIV感染靶细胞的共受体。,细胞因子抑制剂 用于治疗炎症，自身免疫病，移植排斥，休克,细胞因子的临床应用,进入临床实验的细胞因子抑制剂,名称,公司,适应症,临床阶段,可溶性IL-1受体(干粉吸入剂),Immunex,哮喘,I/II期,可溶性IL-1受体（注射剂）,急性髓样白血病,I期,IL-1受体拮抗剂,Amegen,败血性休克(试验中止),类风湿关节炎,完成III期,可溶性IL-4受体,Immunex,哮喘,I/II期,抗IL-4人源化抗体,SmithKline,哮喘,I/II期,抗IL-5人源化抗体,SmithKline,哮喘,II期,可溶性TNF受体II-Fc融合蛋白,Immunex,类风湿关节炎，慢性心衰,上市，I期,可溶性TNF受体I-Fc融合蛋白,Hoffmann-La Roch,休克,类风湿关节炎,多发性硬化症,II/III期,抗TNF单抗（infliximab）,Centocor,Crohns Disease,上市,人源化抗HER2 (EGFR2)单抗,Genentech,乳腺癌转移,上市,DAB389-IL-2(IL-2免疫毒素),Seragen,T细胞淋巴瘤, I型糖尿病,严重类风湿关节炎,牛皮癣,HIV感染,上市I/II期,IL-1ra gene therapy on DBA/1 mice of collagen-induced arthritis,cDNA libraries,PCR,TTTT,TTTT,A,A,+,sequencing,IL-1ra gene,ligate,IL-1ra: IL-1 receptor antagonist,AT clone system (Invitrogen Co.),AT clone vector,Subclone,eukaryotic expression vectors,pcI-neo,EcoRI,EcoRI,EcoRICIP,EcoRI,+,ligation,pcI-IL-1ra,EcoRI,EcoRI,identified,PCRRestriction enzyme,pcDI-IL-1ra,transfection,synoviocyte,COS-7,in vitro,EXPRESSION,IL-1ra protein,ELISAmRNA,pcDI-IL-1ra,in vivo (gene therapy),muscle injection.,DBA/1 mice,type II collagen,induced,DBA/1 mice with arthritis,gene gun,细胞因子的临床应用,已批准上市的细胞因子基因工程药物,正进行临床试验的细胞因子基因工程药物,细胞因子名称,适应症,公司,IL-10,炎症,银屑病,Crohns 病,类风湿关节炎,Immunex，Schering-Plough,IL-12,肿瘤，HIV感染，I型变态反应,GI,Tpo,血小板减少症,GI，沈阳三生,TNF变异体,肿瘤,国内多家企业,Flt3/flk3 Ligand(FL),肿瘤,Immunex,Myeloid Progenitor Inhibitory Factor-1 (MPIF-1),肿瘤大剂量化疗,HGS,TGF-,慢性皮肤溃疡，多发性硬化症,Genzyme,Keratinocyte Growth Factor-2 (KGF-2),促进烧伤、慢性溃疡的伤口愈合；抗癌药物引起的粘膜损伤；炎症性肠道疾病,HGS,MIP-1 变异体,肿瘤化疗的骨髓保护作用,British Biotech,OPG（osteoprotegerin）,骨质疏松,Amgen,Mulitkine（白细胞产生的细胞因子混合制剂）,转移性肿瘤,CEL-SCI,B细胞刺激因子（BlyS）,免疫缺陷,HGS,二、细胞因子的研究热点1、新细胞因子的基因克隆2、细胞因子受体的基因克隆3、细胞因