Presented by Mr. Sharanabasappa Ist M.Pharm. Dept.

of Pharmacology

Subject In Charge Dr. THIPPESWAMY B. S. M.Pharm., Ph.D Professor & Head of the Department DEPT. OF PHARMACOLOGY SSCP, TUMKUR.
SSCP, TUMKUR

 Introduction  Definition  Cytokine categories  General characteristics  Functional classes of cytokines  Chemokines  Therapeutic uses of cytokines  Commercial products of cytokines  References
SSCP, TUMKUR

Cytokines (Greek cyto-, cell; and -kinos, movement) are small cellsignaling protein molecules that are secreted by the glial cells of the nervous system and by numerous cells of the immune system and are a category of signaling molecules used extensively in intercellular communication.
SSCP, TUMKUR

Low molecular weight soluble proteins (polypeptides) produced in response to microbes and other antigens. OR Soluble mediators or glycoproteins that aid in the communication between cells, primary cells of immunological, haematological & neurological systems. NOTE :- Defining cytokines based only on their producer cells or target cells is inaccurate, because one producer cell one target cell has been falsified for almost every cytokine.
SSCP, TUMKUR

About 50 cytokines are identified which are grouped in 6 categories. 1) Interferons 2) Interleukins 3) Tumour Necrosis Factor (TNF, Cachectin) 4) Transforming Growth Factor (TGF) 5) Colony Stimulating Factor (CSF) 6) Growth Factors (PDGF, EGF, FGF, VEGF) Cytokines involved in leucocyte endothelial cell interaction are called Chemokines. Growth factors & other cytokines are named Crinopectins 5
SSCP, TUMKUR

Mediate and regulate the immune system Secretion is brief and self-limited Individual cytokines are produced by multiple cell types Act on multiple cell types-pleiotropic
SSCP, TUMKUR

SSCP, TUMKUR

SSCP, TUMKUR

SSCP, TUMKUR

SSCP, TUMKUR

10

Cytokine actions may be characterized as:

Autocrine (self modulating) Paracrine (modulating cells in the

immediate surroundings)

Endocrine (modulating cells through

circulation in the distant area)
SSCP, TUMKUR

11

SSCP, TUMKUR

12

Cytokines classified according to their biologic actions into three groups: 1) Mediators and regulators of innate immunity - Produced by activated macrophages and NK cells in response to microbial infection - they act mainly on endothelial cells and leukocytes to stimulate the early inflammatory response to microbes
SSCP, TUMKUR

13

2) Mediators and regulators of acquired immunity - Produced mainly by T lymphocytes in response to specific recognition of foreign antigens - They include IL-2, IL-4, IL-5,, IL-13, IFN, Transforming growth factor- (TGF- ) and lymphotoxin (TNF- ) 3) Stimulators of haematopoiesis - Produced by bone marrow, stormal cells, leukocytes - Stimulate growth and differentiation of leukocytes - Stem cell factor, IL-3, IL-7, GM-CSF
SSCP, TUMKUR

14

* Interferons (IFNs): are proteins secreted in

response to viral infections or other stimuli * They include: - INF- produced by leucocytes induced by virus infected cells - INF- produced by fibroblasts - INF- produced by NK cells,TH1 cells, CD8 Tcells
SSCP, TUMKUR

15

Action of INF- and IFN- : - Prevent viral replication - Increase MHC-I expression on viral infected cells helping their recognition by CD8 T-cells - Increase cytotoxic action of Nk cells - Inhibit cell proliferation and tumor growth
SSCP, TUMKUR

16

Action of IFN- : - Activate Macrophages - Increase expression of MHC-I and II on APCs - Enhance cytotoxic actions of Nk cells - Promote production of TH1 and inhibits proliferation of TH2
SSCP, TUMKUR

17

 Group of cytokines (secreted proteins/signaling molecules) that were first seen to be expressed by white blood cells (leukocytes). The function of the immune system depends in a large part on interleukins. The majority of interleukins are synthesized by helper CD4+ T lymphocytes, as well as through monocytes, macrophages, and endothelial cells. They promote the development and differentiation of T, B, and hematopoietic cells.
SSCP, TUMKUR

18

Interleukin IL 1 /IL-1 IL-2 IL-3

Primary Cell Structure Macrophages, NK Cells, B cells T cells T cells T cells T cells T cell & fibroblasts Stromal cells

Primary Activities Inflammation

Activates T cells Haematopoietic growth factor B cell growth Eosinophil & B cell growth Inflammation B & T cell growth

IL-4 IL-5

IL-6 IL-7

SSCP, TUMKUR

19

Interleukin IL-8 IL-9

Primary Cell Structure Macrophages Activated T cells

Primary Activities Chemoattractant for neutrophils T cell growth & Potentiates IgM, IgG & Ig E B cell growth / inhibition of cytokine synthesis by T cells Haematopoietic cofactor Induction of cell mediated immunity B cell growth

IL-10

B cells, T cells

IL-11 IL-12 IL-13

Bone marrow stromal cells Macrophages, B cells T cells

SSCP, TUMKUR

20

Interleukin IL-14

Primary Cell Structure Activated B cells

Primary Activities controls the growth and proliferation of B cells, inhibits Ig secretion T cell & NK cell growth CD4+ T cell chemoattractant Fibroblast stimulation Induces production of IFN , NK cell activity

IL-15 IL-16 IL-17 IL-18

Epithelial cells CD8+ T cells CD4+ T cells Macrophages

SSCP, TUMKUR

21

 Has been described by a variety of different names: Lymphocyte Activating Factor (LAF), Endogenous pyrogen, T cell replacing factor II & B cell differentiation factor, as well as by several other names.  IL-1 represents at least two distinct polypeptides (IL-1 alpha & IL-1 beta).
SSCP, TUMKUR

22

 The MWs of the polypeptides are approximately 17 kDa.  IL-1 has been associated with numerous activities. Some of these include 1. Induction of the IL-2 receptor; 2. Stimulation of pre-B cell differentiation; 3. Augmentation of NK-cell cytotoxicity;
SSCP, TUMKUR

23

4. Induction of adhesion molecules on endothelial cells; 5. Induction of fever; 6. Stimulation of thymocyte proliferation; 7. Enhancement of collagen production; 8. Stimulation of the release of other cytokines involved in haematopoiesis.
SSCP, TUMKUR

24

 Originally described as T cell growth factor (TCGF).  Synthesized & secreted primarily by T cells.  Stimulate the growth, differentiation & activation of T cells, B cells, & NK cells.  Produces its immunological effects by binding to the cellular IL-2 receptor (IL-2R).
SSCP, TUMKUR

25

 Haematopoietic growth factor.  Principle effects are on early haematopoietic progenitors in which Il-3 induces haematopoiesis & cell differentiation.  Administration of IL-3 produces an increase in erythrocytes, neutrophils, eosinophils, monocytes & platelets.  Act synergistically or additively with other haematopoietic growth factors.
SSCP, TUMKUR

26

 Primarily derived from T cells & principle

site of action is the B cell.

 Stimulates B cell proliferation & activation.  Induces IgG1 & IgE expression from B cells

& class II MHC expression.

 Also induces the differentiation of

eosinophils & activity of T cytotoxic cells.

SSCP, TUMKUR

27

 Represents the compounds originally known as T cell replacement factor (TRF), eosinophil differentiation factor (EDF) & B cell growth factor (BCGFII).  Primary effect is on the eosinophilic lineage.  Stimulates eosinophil chemotaxis & eosinophil expansion.
SSCP, TUMKUR

28

 Produced by lymphoid & non-lymphoid cells.  Acts on T cells & B cells.  Stimulates multilineage haematopoiesis, including the maturation of megakaryocytes.  Formerly known as interferonweak antiviral activity.
SSCP, TUMKUR

for its
29

 Acts primarily on pre-B cells to stimulate their differentiation.  Also stimulate the development of human T cells.  Overall, it appears that IL-7 is important in B & T cell development.
SSCP, TUMKUR

30

 Member of a group of glycoproteins known as Chemokines.  Potent chemoattractant for neutrophils.  Has a wide variety of pro-inflammatory effects, including the stimulation of neutrophil degranulation & enhancement of neutrophil adherence to endothelial cells.  IL-8 is the lowest MW IL only 8 kDa.
SSCP, TUMKUR

31

 Group of cytokines which can cause cell death (Apoptosis).  Family members :- 19 cytokines are identified as part of the TNF family on the basis of sequence, functional & structure similarities.  The first two members of the family to be identified were; TNF TNFSSCP, TUMKUR

32

 Monocyte derived cytotoxin; implicated in tumor regression, septic shock & cachexia.  Stimulate cell proliferation & induce cell differentiation under certain conditions.  Potent pyrogen, causing fever by direct action or by stimulation of IL-1 secretion.  Cause cytolysis of certain tumor cell lines.
SSCP, TUMKUR

33

 Also know as lymphotoxin is a cytokine which is inhibited by IL-10.  Lymphotoxin & lymphotoxin , two related cytokines produced by lymphocytes that are cytotoxic for a wide range of tumor cells in vitro & in vivo.

SSCP, TUMKUR

34

 Sometimes referred to as Tumor Growth Factor.  Two classes of polypeptide growth factors, TGF & TGF .  The two classes of TGFs are not structurally or genetically related to one another & they act through different receptor mechanisms.  They dont always induce cellular transformation, and are not the only growth factors that induce cellular transformation.
SSCP, TUMKUR

35

 upregulated in some human cancers. It is produced in macrophages, brain cells, and keratinocytes, & induces epithelial development.

SSCP, TUMKUR

36

 Exists in three known subtypes in humans, TGF 1, TGF 2, and TGF 3.  These are upregulated in Marfan's syndrome and some human cancers.  Play crucial roles in tissue regeneration, cell differentiation, embryonic development, and regulation of the immune system.
SSCP, TUMKUR

37

 colony-stimulating factors are soluble, membrane-bound substances of the hematopoietic microenvironment.  secreted glycoproteins that bind to receptor proteins on the surfaces of hemopoietic stem cells.

SSCP, TUMKUR

38

 Thereby activating intracellular signaling pathways that can cause the cells to proliferate and differentiate into a specific kind of blood cell (usually white blood cells).  They may be synthesized & administered exogenously.
SSCP, TUMKUR

39

 A protein secreted by macrophages, T cells, mast cells ,endothelial cells, and fibroblasts.  Functions as a WBC growth factor.  Stimulates stem cells to produce granulocytes & monocytes.
SSCP, TUMKUR

40

 A naturally occurring substance capable of stimulating cellular growth, proliferation & cellular differentiation.  Usually it is a protein or a steroid hormone.  Growth factors are important for regulating a variety of cellular processes.
SSCP, TUMKUR

41

 Growth factors typically act as signaling molecules between cells. Ex :- Cytokines & hormones which bind to specific receptors on the surface of their target cells.  They often promote cell differentiation and maturation, which varies between growth factors. Ex :- bone morphogenic proteins stimulate bone cell differentiation, while fibroblast growth factors & vascular endothelial growth factors stimulate blood vessel differentiation (angiogenesis)
SSCP, TUMKUR

42

 Individual growth factor proteins tend to occur as members of larger families of structurally & evolutionarily related proteins.  There are many families, like Bone morphogenetic proteins (BMPs) Epidermal growth factor (EGF) Erythropoietin (EPO) Thrombopoietin (TPO) Platelet derived growth factor (PDGF) etc.
SSCP, TUMKUR

43

 Family of chemoattractants for inflammatory cells & include; IL-8 chemotactic for neutrophils Platelet factor 4 chemotactic for neutrophils, monocytes & eosinophils MCP 1 chemotactic for monocytes Eotaxin chemotactic for eosinophils
SSCP, TUMKUR

44

 Chemokines are a family of small (8 to 10 kDa) proteins which act primarily as chemoattractants for specific types of leukocytes.  About 40 different chemokines & 20 different receptors for chemokines are identified.

SSCP, TUMKUR

45

 Classified into four major groups, according to the arrangement of the conserved cysteine (C) residues in the mature proteins. 1. C-X-C chemokines ( chemokines) 2. C-C chemokines ( chemokines) 3. C chemokines ( chemokines) 4. CX3C chemokines
SSCP, TUMKUR

46

 Have one amino acid residue separating the first two conserved cysteine residues.  Act primarily on neutrophils.  IL-8 is typical of this group.  Secreted by activated macrophages, endothelial cells, neutrophils, with limited activity on monocytes & eosinophils.  Inducers are microbial products & other cytokines, mainly IL-1 & TNF.
SSCP, TUMKUR

47

 Have the first two conserved cysteine residues adjacent.  Induces monocyte chemoattractant protein (MCP1), eotaxin, macrophage inflammatory protein - 1 (MIP - 1 ) & RANTES ( regulated & normal T cell expressed & secreted).  Attract monocytes, eosinophils, basophils & lymphocytes but not neutrophils.  Eotaxin selectively recruits eosinophils.
SSCP, TUMKUR

48

 Lack two (first & third) of the four conserved cysteines.  C chemokines (lymphotactin) are relatively specific for lymphocytes.

SSCP, TUMKUR

49

 Contain 3 amino acids between the two cysteines.  Only know member of this class is called fractalkine, which exists in two forms.

SSCP, TUMKUR

50

) Interferon in treatment of viral diseases, cancer.

2) Several cytokines are used to enhance T-cell activation in immunofideficincy diseases, e.g. IL-2, IFN-K,TNF-E. 3) IL-2 and lymphokine activating killer cells (LAK) in treatment of cancer. 4) GM-CSF induces increase in white cell count, it is used: a- To restore leukocytic count after cytotoxic chemotherapy induced neutropenia b- After bon marrow transplantation C- To correct AIDS-associated leukopenia
SSCP, TUMKUR

51

5) Anti-cytokines antibodies in management of autoimmune diseases and transplant rejection: a- Anti-TNF in treatment rheumatoid arthritis & Crohns disease. b- Anti-IL2R to reduce graft rejection 6) Anti-TNF antibodies in treating septic shock 7) Anti-IL-2R E in treating adult T-cell leukemia 8) Anti-IL-4 is under trial for treatment of allergies
SSCP, TUMKUR

52

 ALDESLEUKIN Recombinant human IL-2 (rIL-2), known generically as aldesleukin, is available as Proleukin. Chemical name :- des-alanyl-I, serine-125-human interleukin-2. First approved indication was in 1992 for the treatment of metastatic renal cell carcinoma.
SSCP, TUMKUR

53

 OPRELVEKIN Recombinant human IL-11, available as the product Neumega. Indicated for the prevention of severe thrombocytopenia. Mechanism stimulation & proliferation of both haematopoietic stem cells & megakaryocyte progenitor cells, & the induction of megakaryocyte maturation.
SSCP, TUMKUR

54

 OTHERS IL-3 (muplestim) support of haematopoiesis after high-dose chemotherapy or bone marrow failure, & for myelodysplastic syndromes. IL-6 (sigosix) potential platelet stimulator TNF inhibitors Etanercept, Infliximab etc IL-1 antagonist Anakinra
SSCP, TUMKUR

55

SSCP, TUMKUR

56

 Pathological Basis of Disease. Robbins & Cotran; 7th edition: Pg no. 71-72.  Pharmaceutical Biotechnology. J. A. Crommelin & Robert D Sindelar; 2nd edition: Pg no. 217 to 230.  Text book of Pathology. Harsh Mohan; 5th edition: Pg no. 31-32, 143.
SSCP, TUMKUR

57

 Pharmacology. H. P. Rang, M. M. Dale, J. M. Ritter & P. K. Moore; 5th edition: Pg no. 239-40.  Essential of medical pharmacology. K. D. Tripathi; 6th edition: Pg no. 204-05.  http://en.wikipedia.org/wiki/Cytokine  http://www.cytokines.com/
SSCP, TUMKUR

58

SSCP, TUMKUR

59