第六章 细胞因子 Chapter ６ Cytokines

1. 第六章　细胞因子 Chapter ６ Cytokines

2. Objectives and Requirements • Master the concept of the cytokines; • 2. Familiar with the common properties. • 3. Comprehend the functions of cytokines and the cytokine receptor.

3. 目的、要求 １、掌握细胞因子的概念和细胞因子的命 　　名与分类; ２、熟悉细胞因子的共同特点; ３、了解细胞因子及其受体的功能。

4. 1. History for the research of the cytokines 细胞因子研究简史  In 1957，Interferon （IFN ） was first found  In 1969， termed lymphokine  In1974，denominated cytokine  cytokinology created； international cytokine society founded 《 Cytokine 》journal  Companies for developing cytokines in many countries

5. 细胞因子研究简史  1957年，发现干扰素（Interferon, IFN）； 1969年，提出淋巴因子（lymphokine）的概念 ； 1974年，定名细胞因子（cytokine）；  近二十年来,　细胞因子发展迅猛, 现已发现一百 　　多种细胞因子； 形成专门学说——细胞因子学（cytokinology）； 成立专门学会，如国际细胞因子学会 （international cytokine society）； 创办专门杂志，如《Cytokines》等；  开办了众多细胞因子的研制公司；

6. Mj IL-1 2.　细胞因子的概念　Concept of Cytokine 细胞因子是由免疫细胞和某些非免疫细胞（如血管内皮细胞、表皮细胞和成纤维细胞等）经刺激而合成分泌的一类小分子量可溶性糖蛋白或蛋白多肽。 Cytokines 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. Activated Th1 cell macrophage a A B’ D C P56 B C58 IL-2 105 COOH NH2

7. Cytokine-producing cell Cytokine gene Cytokine Receptor signal Gene activation Biological effects Inducing stimulus Overview of the induction and function of Cytokine 细胞因子诱导与功能概观

8. 3. Production of cytokines 细胞因子的产生 • Normal cells（正常细胞） • Activated lymphocyte, activated monocyte / macrophage, NK cell, fibroblast, epithelial cell, endothelial cell.

9. Tumor cells（肿瘤细胞） • Jurkat cell, WEHI-3, myeloma cell（骨髓瘤细胞） • Genetic engineering technique • 基因工程技术 • Engineering cells, such as E.coli , yeast produce the recombinant cytokines. Cytokine gene E.coli expresses the cytokine protein purification Cytokine （product） E.coli

10. 4.　细胞因子分类　Cytokine classification 　 按细胞因子的结构和功能分６类： Classified based on the structure and function 1. interleukins, ILs （白细胞介素） 2. colony stimulating factor,CSF（集落刺激因子） 3. interferon, IFN（干扰素） 4. tumor necrosis factor, TNF （肿瘤坏死因子） 5. chemokine （趋化因子） 6. growth factor, GF （生长因子）

11. Interleukin, IL　白细胞介素 Interleukins (ILs) are secreted by some leukocytes and act upon other leukocytes, until now, IL-1~IL-35 have been identified. IL-4 IL-13 IL-2

12. Immune bioactivities of IL-2 Mj Mj Th1 NK LAK B Antigen presentation antigen APC IL-1 Th1 Th1 Activation of T cell ( precursor producing IL-2） differentiation Th1 IL-2 reactive cells （Th or TC ） replication g-IFN IL-4 IL-5 IL-6 antigen Th1 Th1 proliferation, differentiation Secretion of antibody

13. Colony stimulating factor, CSF 聚落刺激因子 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, etc G-CSF GM-CSF

14. 集落刺激因子的作用

15. Interferon, IFN 干扰素 The cytokines that have anti-viral infection, anti-tumor, and immune regulation, including IFN, IFN ( type I ); IFN ( type II ). virus infection cell IFN IFN IFN-, IFN-

16. The anti-virus mechanism of IFN- a /  virus viral replication Inhibition of viral replication IFN-induced protein stimulating Signal-transduction nuclear nuclear IFN-a / 

17. Type II interferon, IFN- g

18. Activated Th1 cell NK cell IFN- NK cell B cell macrophage T cell Virus infected cell promoting IL-2 production IL-2R expression Enhancing IV hypersensitivity Plantation rejection cytotoxicity Promoting production of IgG3 and IgG2a Inhibiting Viral replication, proliferation Promoting expression of MHC-II and Fc, antimicroorganism and antitumor, synthesis ofTNF-. Promoting Activation killed Inhibitingexpression of CD23 and production of IgE Immune biological effects of IFN-

19. Tumor necrosis factor,TNF The cytokines that induce directly apoptosis of tumor cells, including TNF, TNF (Lymphotoxin）, FASL, CD70L, CD30L, CD40L, OX40L, TRAIL (TNF related apoptosis-inducing ligand) TNF- 

20. Growth factor The cytokines that promote growth of various cells, epidermal growth factor，insulin-like growth factor，vascular endothelial growth factor vascular endothelial growth factor (VEGF)

21. Chemokine(趋化因子） Chemokines, a group of low-molecular-weight cytokines including interleuki 8, that affect chemotaxis and other aspects of leukocyte behavior. Solution Structure Of The Cx3C Chemokine Domain Of Fractalkine Stromal Derived Factor-1 a

22. 趋化因子引起细胞形态的改变 Chemokines induce cell shape change

23. 5.Biological effects of the cytokines 生物学作用 • Anti-infection and anti-tumor（抗感染，抗肿瘤） • Immune regulation(免疫调节） • Apoptosis（凋亡） • Stimulating the proliferation and differentiation of hemopoietic stem cells（刺激造血干细胞增殖，分化） • Promote the growth and differentiation of various cells • Participation and regulation of the inflammatory responses（参与和调节炎症反应） • Cytokine abnormalities in the pathogenesis in certain diseases （细胞因子异常与疾病）

24. 细胞因子的主要生物学效应 • 抗感染和抗肿瘤作用 • 免疫调节作用 • 参与细胞凋亡 • 刺激造血细胞增埴分化 • 促进各种细胞的生长和分化 • 参与和调节炎症反应 • 细胞异常可导致疾病的发生

27. Circulation 6.main properties of the cytokines　主要特点 （1）action manners　作用方式 Bind to receptors on the membrane of the same cell that secreted it Autocrine　自分泌(自产自销） Binding to receptors on a target cell in close proximity to the producer cells Paracrine　旁分泌（自产旁销） Binding to target cells in distant parts of the body. Endocrine　内分泌（自产远销）

28. B （2）Pleiotropy（多效性） One kind of cytokine may have multiple effects. Activation proliferation differentiation IL-4 proliferation Thymus cell proliferation Mast cell

29. B （3）Redundancy（重叠性） Several kinds of cytokines may have the same or similar effect. Stimulating the proliferation of the B cells IL-2、 IL-4、 IL-5 B （4）synergy（协同性） Cytokines can also act synergistically (two or more cytokines acting together)。 More effectively inducing the exchange of IgE IL-4 + IL-5

30. B （5）Antagonism（拮抗性） Two or more cytokines show opposing activities. _ IL-4 + IFN- IL-4 interdicts the action that IFN-  induces Ig exchanges

31. （6）Rapidity （作用迅速） On stimulation, rapid synthesis , release , action and degragation. Cytokine secretion is a brief, self-limited event. （7）high potency（高活性） Low-concentration cytokines perform remarkable biological effects（pM,10-12 M). （8） Non-specificity(非特异性） perform in the non-specific manner, and not restricted by MHC.

32. （9）binding to specific receptors（结合受体）

33. 7. 细胞因子受体　Cytokine receptors TNF-R 免疫球蛋白超家族受体 I类细胞因子受体 II类细胞因子受体 趋化因子受体 C1C3C2 -S-S- C1C3C2 -S-S- C1C3C2 CC C C -S-S- C1C3C2 G 蛋白 IL-2 IL-3 IL-4 IL-5 IL-6 IL-7 IL-9 IL-11 IL-12 IL-13 IL-15 OSM GM-CSF G-CSF LIF CNTF 生长激素催乳素 IFN-a IFN-b IFN-g TNF-a TNF-bCD40 神经生长因子（NGF） FAS IL-8 RANTES MIP-1 PF4 MCAF NAP-2 IL-1 M-CSF C-kit

34. (2) Soluble cytokine receptors (sCKR) Production:  Most of them break off from the member receptors  Secretory receptor Secretory receptor member receptors

35. ８. Clinical application of Cytokine 细胞因子的临床应用 • Cytokine changes related to diseases • 　（与疾病相关的细胞因子改变）： • deficiency of cytokines and receptors ; • overexpression of cytokines; • high lever of soluble cytokine receptors. • Therapeutic use of cytokines and their receptors(疾病治疗）： • supply and addition； • blockage and antagonist .

36. (1) Common diseases related to cytokines • Infectious diseases（传染病） • tumor • Transplantation rejection(移植排斥反应） • Hematocytopenia（血细胞减少） • Hypersensitivity（超敏反应） • Autoimmune diseases （自身免疫病）

37. 细胞因子引起的病理反应 细胞因子 病理反应 拮抗剂适应症 IL-1 发热,炎症,休克,低血压,-胰岛细胞毒性 炎症性疾病,休克,类风湿关节炎,糖尿病,白血病等 IL-2 发热,渗漏综合征,贫血,低血压 移植排斥,自身免疫病 IL-3 发热,流感样症状,刺激肥大细胞和嗜硷性粒细胞,刺激白血病细胞 IL-4 发热,促进IgE产生,I型变态反应 哮喘等I型变态反应 IL-6 发热,炎症,刺激骨髓瘤细胞 骨髓瘤,炎症性疾病 IL-8 炎症 炎症性疾病 IL-9 哮喘 哮喘 GM-CSF 发热, 流感样症状, 刺激白血病细胞 白血病 TNF 发热,炎症,恶液质,休克 休克, 炎症性疾病,恶液质,类风湿关节炎 IFN 发热, 流感样症状,炎症 炎症性疾病 TPO 骨髓纤维化，微血栓形成 EGF 刺激某些肿瘤细胞增殖 肿瘤 bFGF 刺激某些肿瘤增殖、转移 肿瘤

38. 大剂量TNF注射造成恶液质的小鼠模型

39. 细胞因子与临床治疗 • Clinical use of cytkine inhibitors ① 细胞因子抑制剂的临床应用 IL-1受体拮抗剂（IL-1ra）治疗类风湿关节炎

40. 细胞因子抑制剂 用于治疗炎症，自身免疫病，移植排斥，休克

41. 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 Centocor Crohn's Disease 上市 抗TNF单抗 （infliximab） Genentech 乳腺癌转移 上市 人源化抗HER2 (EGFR2)单抗 DAB389-IL-2 (IL-2免疫毒素) Seragen 上市  I/II期 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

42. DAB389-EGF Seragen 肿瘤 I/II期 人源化抗VEGF单抗 Genentech Inc. 晚期肺癌，肠癌 III期 抗VEGFR2（KDR）嵌合抗体 Albert LoBuglio 转移性肠癌 I期 人源化抗IL-8抗体（ABX-IL8） Abgenix Severe psoriasis I/II期临床 IL13-PE38QQR（IL-13免疫毒素） NeoPharm, Inc 肾癌 I期临床 TNF反义寡核苷酸 Isis 类风湿关节炎，牛皮癣 I期临床 抗eotaxin抗体 （CAT-213） Cambridge Antibody 变态反应 I期临床

43. ② 细胞因子的临床应用(Clinical use of cytokines)

44. 名称 适应症 IFNα 白血病、Kaposi肉瘤、肝炎、癌症、AIDS IFNγ 慢性肉芽肿、生殖器疣、过敏性皮炎、感染性疾病、类风湿关节炎 G-CSF 自身骨髓移植、化疗导致的粒细胞减少症、 AIDS、白血病、再生障碍性贫血 GM-CSF 自身骨髓移植、化疗导致的血细胞减少症、 AIDS、再生障碍性贫血、 Epo 慢性肾衰导致的贫血、癌症或癌症化疗导致的贫血、失血后贫血 IL-2 癌症、免疫缺陷、疫苗佐剂 IFNβ 多发性硬化症 IL-11 放化疗所致血小板减少症 干细胞因子(SCF) 与G-CSF联合应用于外周血干细胞移植 EGF 外用药治疗烧伤、溃疡 bFGF 外用药治疗烧伤、外周神经炎 已批准上市的细胞因子基因工程药物

45. 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 Immunex，Schering-Plough IL-10 炎症,银屑病,Crohn’s 病,类风湿关节炎 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) HGS 肿瘤大剂量化疗 TGF- 慢性皮肤溃疡，多发性硬化症 Genzyme 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 Keratinocyte Growth Factor-2 (KGF-2) 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 免疫缺陷 HGS B细胞刺激因子（BlyS）

46. 9、细胞因子的研究热点 1) 新细胞因子的基因克隆化 cytokine gene cloning 2) 细胞因子受体的基因克隆化 cytokine receptor gene cloning 3) 细胞因子信号转导机制 Signal transduction of cytokines 4) 新一代细胞因子：高活性，多功能，低毒副作用，长 半衰期，高稳定性 New-generation cytokines 5) 细胞因子作为生物应答调节剂（BRM）的临床应用 Clinical application of cytokines as BRM 6) 细胞因子表达调控 Regulation of cytokine expression 7) 细胞因子基因治疗 Gene therapy of cytokines

47. 10. Summary 1. Cytokines are a family of proteins that mediate many of the responses of innate and adaptive immunity. （概念） 2. The same cytokines may be produced by many cell types, and individual cytokines often act on diverse cell types. Cytokines are synthesized in response to inflammatory or antigenic stimuli and usually act locally.（多源性，多效性，协同性） 3. Cytokines often act in an autocrine or paracrine fashion, and may be produced in sufficient quantity to circulate and exert endocrine actions. ( 3种作用方式）. 4. Cytokines mediate their actions by binding with high affinity to receptors. (与受体结合发挥作用）

48. ５.Cytokines serve many functions that are critical to host defense against pathogens.（关键的抗感染因子）. ６. Cytokines provide important amplification mechanisms that enable small numbers of lymphocytes specific for any one antigen to activate a variety of effector mechanisms to eliminate the antigen. （放大效应）. ７. Excessive production or actions of cytokines can lead to pathologic consequences. The administration of cytokines or their inhibitors is a potential approach for modifying biologic responses associated with immune and inflammatory diseases. (与临床密切相关）

49. 11. Practice Quiz 1. Cytokines may exhibit __________ action, signaling the cells that produce them. a. antagonistic b. autocrine c. endocrined. paracrine. e. synergistic 2. Cytokines are NOT a. antigen specific.b. capable of activating more than one cell type.c. made by lymphocytes.d. small protein molecules. e. synthesized de novo in response to antigen or other cytokines. 3. Several cytokines may have the same effect on the cells they bind. This is an example of a. a cascade.b. antagonism.c. pleiotropism.d. redundancy. e. synergy.

50. 4. Characterization of cytokine activities is NOT made more difficult by their a. gene structure.b. pleiotropism.c. redundancy.d. secretion close to target cell membranes.e. short half-lives. 5. Interferons a. activate B cells to make virus-specific antibodies.b. are Th2 cytokines.c. are virus proteins that interfere with activation of cytotoxic T cells.d. block virus infection of host cells. e. inhibit virus replication by infected cells. 6. A cytokine can do all of the following EXCEPT a. bind to receptors which do not share cytokine-binding subunits.b. bind to its specific receptor on the same cell that produced it.c. bind to receptor antagonists produced by pathogenic viruses.d. compete with other cytokines whose receptors share signal-transducing subunits e. upregulate (increase) synthesis of high affinity subunits for its receptor.