cytokines, surface molecules, antibodies jan novák
TRANSCRIPT
CYTOKINES, SURFACE MOLECULES, ANTIBODIES
Jan Novák
The Immune System is the Third Line of Defense Against
Infection
Clonal Selection of B Cells is Caused by Antigenic
Stimulation
• Humoral Immunity - apoptosis
Programmed cell death (“Falling away”). Human body makes 100 million lymphocytes
every day. If an equivalent number doesn’t die, will develop leukemia.
B cells that do not encounter stimulating antigen will self-destruct and send signals to phagocytes to dispose of their remains.
Many virus infected cells will undergo apoptosis, to help prevent spread of the infection.
• Immunological Memory– Antibody Titer: The amount of antibody in the
serum.– Pattern of Antibody Levels During Infection– Primary Response: After initial exposure to antigen, no antibodies
are found in serum for several days. A gradual increase in titer, first of IgM and then
of IgG is observed. Most B cells become plasma cells, but some B
cells become long living memory cells. Gradual decline of antibodies follows.
Monomer: A flexible Y-shaped molecule with four protein chains: 2 identical light chains 2 identical heavy chains
Variable Regions: Two sections at the end of Y’s arms. Contain the antigen binding sites (Fab). Identical on the same antibody, but vary from one antibody to another.
Constant Regions: Stem of monomer and lower parts of Y arms.
Fc region: Stem of monomer only. Important because they can bind to complement or cells.
•Antibody Structure
Antibodies are Proteins that Recognize Specific Antigens
• Consequences of Antigen-Antibody
Binding– 1. Agglutination: Antibodies cause antigens
(microbes) to clump together. IgM (decavalent) is more effective that IgG (bivalent). Hemagglutination: Agglutination of red blood cells.
Used to determine ABO blood types and to detect influenza and measles viruses.
2. Opsonization: Antigen (microbe) is covered with antibodies that enhances its ingestion and lysis by phagocytic cells.
• Humoral Immunity• 3. Neutralization: IgG inactivates viruses by
binding to their surface and neutralize toxins by blocking their active sites.– 4. Antibody-dependent cell-mediated
cytotoxicity: Used to destroy large organisms (e.g.: worms). Target organism is coated with antibodies and bombarded with chemicals from nonspecific immune cells.
– 5. Complement Activation: Both IgG and IgM trigger the complement system which results in cell lysis and inflammation.
Consequences of Antibody Binding
• Immunoglobulin Classes• I. IgG Structure: Monomer Percentage serum antibodies: 80% Location: Blood, lymph, intestine Half-life in serum: 23 days Complement Fixation: Yes Placental Transfer: Yes Known Functions: Enhances phagocytosis, neutralizes toxins
and viruses, protects fetus and newborn.
• Immunoglobulin Classes• II. IgM Structure: Pentamer Percentage serum antibodies: 5-10% Location: Blood, lymph, B cell surface (monomer) Half-life in serum: 5 days Complement Fixation: Yes Placental Transfer: No Known Functions: First antibodies produced during an
infection. Effective against microbes and agglutinating antigens.
• Immunoglobulin Classes• III. IgA Structure: Dimer Percentage serum antibodies: 10-15% Location: Secretions (tears, saliva, intestine, milk), blood and
lymph. Half-life in serum: 6 days Complement Fixation: No Placental Transfer: No Known Functions: Localized protection of mucosal surfaces.
Provides immunity to infant digestive tract.
• Immunoglobulin Classes• IV. IgD Structure: Monomer Percentage serum antibodies: 0.2% Location: B-cell surface, blood, and lymph Half-life in serum: 3 days Complement Fixation: No Placental Transfer: No Known Functions: In serum function is unknown. On B cell
surface, initiate immune response.
• Immunoglobulin Classes• V. IgE Structure: Monomer Percentage serum antibodies: 0.002% Location: Bound to mast cells and basophils throughout
body. Blood. Half-life in serum: 2 days Complement Fixation: No Placental Transfer: No Known Functions: Allergic reactions. Possibly lysis of
worms.
CYTOKINES
a diverse group of non-antibody proteins released by cells that act as intercellular mediators, especially in immune processes
Low molecular weight soluble proteins (polypeptides) produced in response to microbes and other antigens
They act via cell surface receptors to mediate and regulate the amplitude and duration of the immune-inflammatory responses, through activation of macrophages, controlling growth and differentiation of T and B cells
CYTOKINES
Naming of Cytokines
1. Monokines - produced by mononuclear phagocytes (monocytes)
2. Lymphokines - produced by activated T cells, primarily helper T cells
3. Interleukins - cytokines made by one leukocyte and acting on other leukocytes 4.Chemokines-cytokines with chemotactic activities
Classification of Cytokines Interleukins - cytokines made by one leukocyte and
acting on other leukocytes Chemokines - cytokines with chemotactic activities Interferons – tissue antiviral cytokines Transforming growth factors – growth, regulation Colony stimulating factors – growth, stimulation Tumor necrosis factors – induction of apoptosis
Properties 1.Produced by cells involved in both natural and specific
immunity 2. Mediate and regulate immune and inflammatory
responses 3. Secretion is brief and limited not stored as pre-formed
molecules. Synthesis is initiated by new short-lived gene transcription mRNA is short-lived
5. Redundancy -similar functions can be stimulated by different cytokines. Receptors for cytokines are heterodimers (sometimes heterotrimers) that can be grouped into families in which one subunit is common to all members of a given family. (hard clinical diagnostics)
6. Often influence the synthesis of other cytokines. They can produce cascades, or enhance or suppress production of other cytokines. They exert positive or negative regulatory mechanisms for immune inflammatory responses
7. Often influence the action of other cytokines. antagonistic -cytokines causing opposing activities additive synergistic -two or more cytokines acting together (greater than
additive)
Properties
8. Bind to specific receptors on target cells with high affinity.. 9. Cellular responses to cytokines are generally slow (hours),
require new mRNA and protein synthesis
Properties
Cytokine action can be
Functional Categories of Cytokines1) Proinflammatory cytokines
- Produced by activated microphages and NK cells in response to microbial infection
- chemotaxis- killing- IL1, IL-6, IL-8, IL-12, IL-18, TNF-a
Functional Categories of Cytokines2) Anti-inflammatory cytokines
- Produced mainly by T cells- IL1Ra, IL-4, IL-10, TGF-b
Functional Categories of Cytokines3) Growth factors
- IL2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-11, IL-14, IL-15, G-CSF, EPO…
Functional Categories of Cytokines4) Cytokines of humoral immunity
- IL-4, IL-5, IL-9, IL-13
Functional Categories of Cytokines5) Cytokines of cellular immunity
- IL-2, IL-12, IFN-g
Functional Categories of Cytokines6) Cytokines with antiviral potential
- IFN-a,b,g,
Cytokine Receptors
Divided into several families based on their structure and activities
Hematopoietin family receptors are dimers or trimers conserved cysteines in their extracellular
domains and a conserved Trp-Ser-X-Trp-Ser sequence.
Examples are receptors for IL-2 through IL-7 and GM-CSF.
Interferon family receptors have the conserved cysteine
residues but not the Trp-Ser-X-Trp-Ser sequence,
Eg. the receptors for IFNa, IFNb, and IFNg.
Tumor Necrosis Factor family receptors have four extracellular domains;
they include receptors for soluble TNFa and TNFb as well as membrane-bound CD40 (important for B cell and macrophage activation) and Fas (which signals the cell to undergo apoptosis).
Chemokine family receptors have seven transmembrane helices
and interact with G protein. This family includes receptors for IL-8, MIP-1 and RANTES.
Chemokine receptors CCR5 and CXCR4 are used by HIV to preferentially enter either macrophages or T cells.
Therapeutic Uses of Cytokines1) 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-,TNF-
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
5) Anti-cytokines antibodies in management of
autoimmune diseases and transplant rejection:
a- Anti-TNF in treatment rheumatoid arthritis
b- Anti-IL2R to reduce graft rejection
c- Anti-TNF antibodies in treating septic shock
d- Anti-IL-2R in treating adult T-cell leukemia
e- Anti-IL-4 is under trial for treatment of allergies
Adhesive molecules
Adhesion of cells Signal transduction
Groups of adhesive molecules Integrins
Structures of intercellular matrix Formed by two subunites (a and b) Quiscent and activated conformation LFA-1, VLA-1-5, CR3, CR4
Groups of adhesive molecules Molecules of immunoglobulin structure
ICAM, CD80/86, CD2
Groups of adhesive molecules Selectins and Lectins
L (leucotytes) E (endothelial) P (platelet) Interactions between leukocytes and endothelium Receptors of NK cells (Lektins of C type)
Groups of adhesive molecules Mucins
CD43 Supression of interlekcocyte contact
FC receptors
Bind FC parts of immunoglobulins Divided according to the class of antibody
they bind CD16, CD32, CD64
Complement receptors
Phagocytosis of opsonized particles (CR3,4) Clearance of immunocomplexes Receptors for chemotaxis (C3aR, C5aR)
CD – cluster of differentiation system was intended for the classification of
the many monoclonal antibodies generated by different laboratories around the world against epitopes on the surface molecules of leukocytes
proposed surface molecule is assigned a CD number once two specific monoclonal antibodies are shown to bind to the molecule
+/- or hi, mid or low (alternatively bright, mid or dim
CD – cluster of differentiation CD1, CD2, CD3, CD4, CD5, CD8, CD11,
CD14, CD16, CD19, CD20, CD25, CD28, CD34, CD38, CD45, CD56, CD69, CD96, CD161