1. Cytokines Dr. habil. Kőhidai László Assoc. Professor Dept. Genetics, Cell- and Immunobiology Semelweis University Chemotaxis - Elective Course 20 10/2011 – EM-ED

2. Cytokines - Definition ”Cytokine” (Cohen 1974):”Cytokine” (Cohen 1974): Molecules capable for chemical communication between cells;Molecules capable for chemical communication between cells; Most of them are growth factors or differentiation inducers;Most of them are growth factors or differentiation inducers; Regulators of embryogenesis, haematopoesis, tissue ‘repair’, inflammatory and immune functions. E.g. normal growth and development, wound healing.Regulators of embryogenesis, haematopoesis, tissue ‘repair’, inflammatory and immune functions. E.g. normal growth and development, wound healing. Lymphokines, monokines, chemokinesLymphokines, monokines, chemokines

3. Characterization of cytokines Summary Proteines / glycoproteines Work in solved form Low M.W. Inducibility Effects elicited on low cc. Narrow spectrum Specific receptors Network Multifunctionality Synergy Progression factor Competency factor

4. Grouping - Functions Natural immunity and inflammation Natural immunity and inflammation (IFN, TNF, IL-1, IL-6, chemokines) Regulating of lymphocyte activity, Regulating of lymphocyte activity, differenctiation differenctiation (IL-2, IL-5, IL-6) Maturation of immune cells Maturation of immune cells (IL-3, GM-CSF)

5. Characteristics Low M.W. 15-25kDa Products of leukocytes – BUT other cells can also produce them Production induced by  infectious substances and toxins (e.g. LPS)  tissue damage (thrombocyte aggregation)  immune iductions (stages of hypersensitivity)  other cytokines e.g. IL-1 (pro-inflammatory cytokines) Constitutive – e.g. IL-16, IL-18, EPO, M-CSF

6. Majority of cytokines are released by the cells before action BUT there are  membrane bound forms (TNF),  pooled in i.c. vesicles i.c. (mast cells vagy thrombocyte)  stored in extra-cellular matrixban (TGF-beta) – rapid release and action Narrow action radius  autocrine – cytokine produced by the cell acts on the producer cell  paracrine – cytokine acts on the neighbour cells of the producer cell  endocrine – cytokine acts on cells far from the producer cell IL-1 from site of injury to thalamus – regulation of temperature IL-6 from site of injury to liver – acut phase reaction Erythropoetin from kidney to bone marrow – RBC differentiation Characteristics

7. Autocrine Paracrine T cell Macrophage Cytotoxicity ^ Monokine prod. ^ Adhesion mol. ^ Activation ^ IL-2R ^ Lymphokines ^ Endocrine Brain Prostaglandins Pain threshold Fever IL-1

8. Characteristics Effects on low concentrations : 10 -12 – 10 -9 M 10 IL-1 molecules are enough to induce IL-2 synthesis in T cells ! 50 IFN gamma molecules are enough to induce anti-viral effects !

9. Receptors Specific receptors: high affinity, inducibility, specific intracellular signal-transduction and second messenger pathways.

10. Cytokine networks: cytokines are never produced alone, their synthesis is controlled by the balance of positive and negative signals.

11. inflammation Defense against infections Immune- suppression Autoimmunity, rejection

12. Characteristics Synery : combined effetc of two or more cytokines is higher than the simple summary of the individual effects blockAntagonism : cytokines can block each other e.g. IL-4 and IFN- . Multifunctionality: (pluripotent, pleiotrope) more, different effectsdiverse target cells several cytokines possess more, different effects on the diverse target cells Progressive factor: proliferation cytokines induce cell proliferation (IL-2, IL-3, IL-5) Competency factor: differentiation inducing of differentiation (IL-1, IL-4, IL-6)

13. Factors (names) describe the function - Historical  colony stimulating factor CSF  tumor necrosis factor TNF  transforming growth factor TGF Nomenclature Classification based on characteristic gene sequence – result of joint agreement  IL-1 – IL-24 Names describing the structure and function  chemotactic factors - ‘chemokins’.

14. Regulation DNA-level  genes turned on continously e.g. M-CSF, G-CSF, SCF, IL-6, Epo génjei  other genes of cytokines are turned off  co-ordinated expression e.g. IL-3, IL-4, IL-5, GM-CSF genes on 5. Chrs q Post-transcription level Post-secretion level  short half life time in serum e.g. TNF = 15min  soluble cytokine receptors, e.g. sTNF-R Target cell  up- and down-regulation of receptors  induction or amplification of inflammation by i.e. pro- inflammatory cytokines

15. Examples cells possessing nucleus IL-1 – produced by all cells possessing nucleus, its spectrum is wide, it has several target cells. In vivo effects: hypotensio, fever, loss of weight, neutropoenia and acut phase reaction. dendritic cells The chief function is elicited as a factor released from dendritic cells. Promotes proliferation of T lymphocytes Promotes proliferation of T lymphocytes. inducerof colony formationin IL-3 – haematopoetic growth factor, inducer of colony formation in erythroid, megakaryocyte, neutrophil, eosinophil, basophil, mast cell and monocyte cell lines.

16. Examples multifunctional cytokine IL-6 – multifunctional cytokine, produced by lymphoid and B-cell funkcions haematopoesisacut phasis reakction non-lymphoid cells. Regulator of B-cell funkcions, haematopoesis and the acut phasis reakction. inflammation IL-8 - cytokine released in inflammation, several cells can produce it, chemoattractant It works as chemoattractant in neutrophils. Can also influence activity of basophils and some subpopulations of lymphocytes. angiogen Effective angiogen factor.

17. Examples intracellular pathogens. IL-12 – important in responses against intracellular pathogens. IFN gamma synthesis in T-cells and in NK cells. Inducer of IFN gamma synthesis in T-cells and in NK cells. NK and ADCC activity. Increases the NK and ADCC activity. proliferation and differentiation Inducer of proliferation and differentiation in Th1 CD4 cells.

19. Clinical significance auto-immune diseases infections immune-defficiencies malignant tumors