ANTIGEN, ANTIBODY AND COMPLEMENT

Antigen
Antigens are simple or complex foreign substances, which may be organic, inorganic or biological agents and they
Enhances Provokes Elicits

the immune response when they enters in the body parentally.

Antigen
The word Parenteral (par- beyond) and (Enteron git) is used in definition because orally administered antigens are usually denatured by digestive enzymes and their antigenicity is destroyed. When given parenterally it do not undergo any inactivation and can initiate antibody production.
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Antigen
When a foreign bodies or substances enter the system or body, it mediates
Immediate or Delayed type of immune response.

Various events of defense mechanism will be taking place to remove the antigen. Humoral and cell mediated immunity will taken place at the last.
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Classification of antigen
Complete antigens or Immunogens :
When these antigens enters the body evokes the immune response without any assistant or carrier molecule .They possess both qualities
Immunogenicity and Antigenicity.

Incomplete antigens or Haptens :

These are the foreign substance they require carrier molecule to act as a complete antigen. Such antigens are called as incomplete antigens or Haptens.
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Haptens
Haptens are low molecular weight compounds which are having antigenic property but lacks immunogenic property.
The immunogenic property or production of antibody is governed by the carrier molecule. The carrier molecule is a non-antigenic component and helps in provoking the immune response.
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Haptens
A hapten is equipped with chemically reactive side chains such as
Azide, Sulphonates, Arsinate and Carboxyate etc.

Antibodies are also raised against these groups also. Normally the adjuvant are used as a carrier compound for haptens and making it as complete antigens.
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Adjuvants:
It is a chemical, which when administered with the antigens, enhances or provokes the immunity.

Adjuvants are chemical suspension or liquid suspension in which antigen or foreign proteins are dissolved. Since most of the antigens are proteineous in nature, they exhibit a maximum antigenicity if injected simultaneously with the suitable adjuvants.
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Adjuvants
The commonly used adjuvants are:
Freunds complete adjuvants with lipid suspension with mycobacterium in it. Freuds incomplete adjuvants are lipid suspension or alum suspension without any mycobacterium particles.

These adjuvants are enhances the activation of B and T lymphocytes and macrophages. Hence it has tremendous importance in the vaccine production and injection. ex: Hepa B vaccine yeast based (causes allergic reactions) adjuvants
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Adjuvants
Adjuvants can react in Several Ways: 1. Alter the distribution and persistence of antigen within the positive host. 2. Stimulate lymphocytes production nonspecifically. 3. Activate macrophages.

4. Alter traffic of circulating lymphocytes.

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Types of antigens
Exogenous antigens :
These antigens enters the body or system and start circulating in the body fluids and trapped by the APCs (Antigen processing cells such as macrophages, dendritic cells etc.)

The uptakes of these exogenous antigens by APCs are mainly mediated by the phagocytosis.
Ex: bacteria, viruses, Fungi etc.
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Types of antigens
Endogenous antigens :
These are bodys own cells or sub fragments or compounds or the antigenic products that are produced.

These are further classified into a. Autoantigens : These are synthesized by the body. Ex: nucleoproteins, nucleic acids etc. b. Alloantigens : Same set of molecules with the genetic variation. Ex: blood group antigens. HLA (Histocompatibility Leukocyte antigens) etc.
The endogenous antigens are processed by the macrophages which are later accepted by the cytotoxic T cells.
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Classification; according to its relationship to the host

1. Autologous within the same individual 2. Syngeneic within individual with inbred strain (identical twins) 3. Allogeneic or homologous within the same species 4. Xenogeneic or heterologous across species boundary
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Classification: according to its presence in the host

1. Sequestered autologous substance inaccessible to Ab forming tissues ex: CNS, cornea, sperm 2. Tissue type Ag genetically endowed, naturally present in the tissues 3. Tissue specific Ag unique to a particular organ ex: prostate specific Ag (tumor marker in prostate
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Classification: according to ability to stimulate response

1. Thymus dependent require of T helper -Activates both CMI and HMI, provokes memory 2. Thymus independent activate B cells directly -Only IgM Ab are produced Ab in Ag ogen

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Classification: according to serologic behaviour

1. Agglutination particulate 2. Precipitinogen soluble

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Chemical Nature of Immunogens/ Antigens

A. Proteins -The vast majority of immunogens are proteins. These may be pure proteins or they may be glycoproteins or lipoproteins.
In general, proteins are usually very good immunogens

B. Polysaccharides
Pure polysaccharides and lipopolysaccharides are good immunogens.
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Chemical Nature of Immunogens/ Antigens

C. Nucleic Acids - Nucleic acids are usually poorly immunogenic. However, they may become immunogenic when single stranded or when complexed with proteins.
D. Lipids - In general lipids are nonimmunogenic, although they may be haptens.
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Property of antigens/ Factors Influencing Immunogenicity

1. Molecular mass: Smaller molecules does not provoke immune system.
The antigens should possess an optimum molecular mass or large molecule which then binds with the receptors and provoke the immune response. The molecular weight should be between 1000 to 10,000.

2. Antigenic determinant size: Antigenic determinants or epitopes are the regions of antigen which specifically binds with the antibody molecule.
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Property of antigens/ Factors Influencing Immunogenicity

3.Foreignness - The immune system normally discriminates between self and non-self components such that only foreign molecules are immunogenic.
4. Chemical Composition - In general, the more complex the substance is chemically the more immunogenic it will be. 5. Physical form - In general particulate antigens are more immunogenic than soluble ones and denatured antigens more immunogenic than the native form.
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Property of antigens/ Factors Influencing Immunogenicity

6. Rigidity maintenance of the conformational structure of the antigen - Loss/alteration/denaturation/ leads to loss of reactivity 7. Insolubility more particulate and insoluble more immunogenic

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Property of antigens/ Factors Influencing Immunogenicity

8. Genetic Factors - Some substances are immunogenic in one species but not in another. Similarly, some substances are immunogenic in one individual but not in others (i.e. responders and non-responders). The species or individuals may lack or have altered genes that code for the receptors for antigen on B cells and T cells They may not have the appropriate genes needed for the APC to present antigen to the helper T cells.
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Property of antigens/ Factors Influencing Immunogenicity

9. Age - Age can also influence immunogenicity. Usually the very young and the very old have a diminished ability to elicit and immune response in response to an immunogen.

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Structure of Antigen

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Antigen molecule
1. Univalent, unideterminant 2. Multivalent, unideterminant 3. Multivalent, multideterminant

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Factors contributing to immune responsiveness

Dosage of the antigen Frequency of encounter Route of introduction to the host Age and gender Genetic endowment Underlying diseases and medication

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Antibodies (Ab)
Glycoproteins that bind antigens with high specificity and affinity. Produced in B cells in response to an antigen: Activated B cells => Plasma cells => secretion antibodies Found in plasma and various body fluids Sometimes are called gamma globulins or immunoglobulins Five classes: IgG, IgM, IgA, IgE, IgD Can be isolated in the gamma-globulin fraction of protein by electrophoresis

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Basic Structure
4 polypeptide chain unit (Y shaped) 2 heavy chains (five kinds: , , , , ) 2 light chains (two types: , ) Disulfide bonds - Inter-chain - Intra-chain
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 The secretions that make up the tips of the Ys arms vary greatly from one antibody to another, this is called the variable region. It is these unique contours in the antigen-binding site that allow the antibody to recognize matching antigen

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Basic Structure
Domains V L & CL VH & CH1, CH2, CH3 (or CH4) Functional Properties: VL & VH: Bind to Ag CL , CH1 : Genetic marker CH2 : Complement binding site; Placental transfer (IgG) CH2/ CH4 : binding to FcR complement binding site
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Valance and avidity

Valence: the maximum of antigenic determinants with which antibody can react (e.g. IgG: 2,Fab:1) Avidity: the firmness of association between a multideterminant antigen and antibodies produced against it Having multiple binding sites for antigen means multi-valence and can increase its avidity antigens on particles such as bacteria or virus

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Bifunctional Antigen binding funciton Effector functions

Specificity Binds to specific antigen

Cross - Reactivity Similar to the determinant of antigen

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Affinity Tightness of the FIT

Avidity Overall strength

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Human Ab Classes
IgG Gamma () heavy chains
4 subclasses (IgG1, IgG2, IgG3, IgG4) have slightly different sequences in Hchain and corresponding differences in functional activities.

IgM Mu () heavy chains IgA Alpha () heavy chains

2 subclassses (IgA1, IgA2)

IgD Delta () heavy chains IgE epsilon () heavy chains

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IgG
Distribution Vascular intra and extra

IgM
Vascular intra

IgA
Vascular (intra)++ Mucosa

IgD

IgE

Membrane FccR on of (naive mast cells, cells) basophils

Subclasses

2
+++

Breast Milk/ + Colostrum Placental transfer ++

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IgG
Predominant Immunoglobulin in humans 70-75% of total Ig pool High diffusion coefficient allowing it to enter extravascular spaces more rapidly 7S Molecule 150,000 MW

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IgG
Properties: Found in vascular and extravascular spaces, secretions Provide immunity to most blood borne infectious agents Major antibody if the secondary immune response The only Ab class that can cross placenta *Ig3 more disulfide bonds

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IgG

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IgA
Structure
Serum: monomer Secretions (secretory IgA, sIgA):
Dimer J chain (joining chain) SP (secretory piece) (involved in the transepithelial transport of exocrine IgA and stabilizes IgA againt proteolytic degradation)

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IgA
Properties
Found mainly in serum, external secretions (such as: colostrum, milk, saliva) 2nd highest serum Ig Major secretory Ig A first line of defence against microbial invaders at mucosal surfaces

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 Structure

IgM

Pentamer Extra domain (CH4) J chain (joining chain)

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IgM
Properties
Found primarily in plasma, the surface of B cell 3rd highest serum Ig, <10% of serum Ig First Ab produced by fetus and B cells Agglutinating Ig Binds Fc receptor B cell surface Ig

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IgD
Structure Monomer Tail piece Propeties 4th highest serum Ig B cell surface IG

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IgE
Structure: Monomer Extra domain (CH4) Properties Least common serum Ig Associated with acute inflammation, parasite infection and allergic reactions (such as asthma)

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Monoclonal antibodies (mAb)

Polyclonal antibodies Monoclonal antibodies:
Monospecific Are standard research reagents and have significant clinical utility

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Innate Immune System

Immune System
Adaptive Immune System

Complement

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Complement
Definition: Complement is a group of serum proteins at least 20 in number, which normally remain inactive form and when activated they act in concert in an orderly sequence to exert their biological effects

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Production of complement components:

Complement are synthesized by the liver, spleen, macrophage and intestinal epithelial cells.

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Inactive Complement

C1, INH, Factor 1, H, c4-bp

Bacterial Invasion

Activation of Complement
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PROTEINS OF THE COMPLEMENT SYSTEM

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C1
C1 in blood serum as a molecular complex containing:
6 molecules of C1q 2 molecules of C1r 2 molecules of C1s

The constant regions of mu chains (IgM) and some gamma chains (IgG) contain a binding site for C1q
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C3
C3 is the most abundant protein of the complement system (~1.3 mg/ml) Ability to activate itself

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C5
Unique to the terminal complement pathway Present in plasma at 70 mg/L Together with the other proteins of the terminal pathway, C5b forming the MAC (C5b6789)

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Properdin
Constituent of normal serum with a concentration of approx. 5-15 ug/ml Stabilizes the C3 convertase

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Factor D
Plasma protein that circulates in active enzyme form Cleaves Factor B

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Alternative Pathway
Serum Protein MW, kD Factor B 93 Concentration Function ug/ml 200 Binds to C3b to form C3 convertase 1-2 Cleaves Factor B 25 Stabilizes C3bBb-C3 convertase

Factor D

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Properdin

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MBL
Binds to mannose or related sugars to initiate the pathway An acute phase protein due to its presence in the serum but increases during initial inflammatory responses

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Fluid Phase Regulators

C1 inhibitor or C11NH Molecular weight of 150,000 Inactivate C1 by binding to the active site of C1r and C1s Inhibits clot formation essentials Factor H MW of 160,000 Prevents binding of Factor B by binding C3b Allows factor 1 to break down C3b
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Factor I A serine protease that inactivates C3b and C4b when they are associated with Factor H C4 binding protein or C4BP Cofactor for Factor I in the activation of C4b Abundant in plasma Causing cessation of the classical pathway

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S protein Control protein that acts at a further level complement activation Also known as VITRONECTIN

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