I.

Definition of antigen
Antigen is substance which when introduced parentally into the body stimulates the production of an antibody with which it reacts specifically and in an observable manner Antigen: Immmunogen Tolerogen Allergen Vaccine

1. Immunogen: the antigen that induce specific immune

response

Microbes; bacteria ,virus; fungi and parasites xenoantigenei or allogeneic tissues or organs: grafted skin , bone marrow

2.Tolerogen: antigen that induce Immunologic tolerance

Immunologic tolerance is unresponsiveness to an antigen that is induced by prior exposure to that antigen.

tolerogen

3. Allergen: antigen that induce Anaphylaxis (severe immediate hypersensitivity reaction occurring as a result of rapid generalized mastcell granulation)

Allergen: some medicine, flower powder, seafood

4. Vaccine: antigens that induce a protection immune response against microbes and are used to prevent diseases Killed vaccine: Rubella virus Attenuated vaccine: Measles Toxoid :Tetanus

Based on Immunogenicity
Complete antigen : substances with both immunogenecity and immunoreactivity
By convention , we call complete antigen as antigen. Incomplete antigen (hapten): substances only with immunoreactivity

Hapten +carrier

complete antigen (immunogens)

Hapten: Only possess immunoreactivity Carrier: Make hapten obtain the immunogenicity

Based on Chemical nature

Proteins
Majority of immunogens are proteins (pure proteins or they may be glycoproteins or lipoproteins). Proteins are usually very good immunogens.

Polysaccharides
Pure polysaccharides and lipopolysaccharides are good immunogens.

Nucleic Acids
Nucleic acids are usually poorly immunogenic. However, they may become immunogenic when single stranded or when complexed with proteins.

Lipids
In general lipids are non-immunogenic, although they may be haptens.

According to source of antigens Xenoantigen Alloantigen Autoantigen Heterophile antigen

(1) Xenoantigen
An antigen that is found in more than one species. An antigen is something that is capable of inducing an immune response. The prefix "xeno-" means foreign or other. It comes from the Greek "xenos" meaning stranger, guest, or host. Pathogens: bacteria, virus , fungi, parasite Exotoxin and toxoid Exotoxin

Produced by G+ bacteria
Strong antigenicity and pathogenicity Toxoid : exotoxin that loses its toxicity but maintains its antigenicity under suitable conditions (low concentration of formaldehyde ) Such as tetanus toxoid , diphtheria toxoid

bacteria

Pathogens

Fungi

HIV

Heterophile Ag (forssman Ag)

Common Ags shared by different species ( between human and animal or microbes, between different species of microbe) (eg) M protein of streptococus bears common antigen determinant with basement membrane of kidney
(This common between bacteria and human being can causes poststreptococcal glomerulonephritis)

No specificity of species

Significance . immunopathology
. Diagnosis

(2) Alloantigen
Antigens of red blood cell ABO system (blood typing) - very important in transfusion Rh system (Han race :>99%Rh+)----haemolytic disease of

the newborn (HDNB)

HLA system (Human leukocyte antigen) - relate to transplantation - very important in immune regulation

ABO system
Blood typing A B AB O antigen of RBC A B A,B antibody in serum anti-B anti-A anti-A, anti-B

(3) Autoantigen
Release of sequestered Ag

Lens protein is released into blood to induce immune response

to induce inflammation of lens Change of molecular structure of auto-tissues Denatured IgG becomes antigen to induce production of antibody ( rheumotoid factor)

In patient suffering from rheumotoid arthritis

II. According to whether need the help of T cells when B cells produce Ab

TD-Ag (thymus dependent Ag ) TI-Ag (thymus independent Ag)

1.TD-Ag (thymus dependent Ag )

TD-Ag can stimulate B cell to produce Ab with the help of T cell The most of TD-Ag belong to protein many kinds of determinants stimulate B cell to produce :IgG, IgM, IgA capable of inducing CMI immune memory

2. TI-Ag (thymus independent Ag)

TI-Ag can stimulate B cells to produce Ab without the help of T cell
most are polysaccharide

more ,same, repeat determinant only induce B cell to produce IgM can not induce CMI no immune memory

SUPERANTIGENS
When the immune system encounters a conventional T-dependent antigen, only a small fraction (1 in 104 -105) of the T cell population is able to recognize the antigen and become activated (monoclonal/oligoclonal response). However, there are some antigens which polyclonally activate a large fraction of the T cells (up to 25%). These antigens are called superantigens

Eg: Staphylococcal enterotoxins (food poisoning), Staphylococcal toxic shock toxin (toxic shock syndrome), Staphylococcal exfoliating toxins (scalded skin syndrome) and Streptococcal pyrogenic exotoxins (shock). The diseases associated with exposure to superantigens are, in part, due to hyper activation of the immune system and subsequent release of biologically active cytokines by activated T cells.

 Tumor specific Ag ( TSA)

Only expressed on the tumor cells but normal cells The tumor antigens encoded by genomes oncogenic virus EB virus ---B cell lymphoma HPV-cervical carcinoma Tumor associated Ag (TAA) Highly expressed on tumor cells but lowly normal cells, such as AFP CEA expressed on

AFP (alpha-fetoprotein): over-expression in liver cancer CEA (carcinoembryonic antigen): over-expression in carcinoma of colon , pancreas, stomach ,and breast

DEPEND ON SIZE
Very high molecule haemocyanin (MW 6.75million) highly antigenic Low molecular weight molecule (<10,000) non antigenic or feeble They render antigenicity by absorbing inert particles like bentonite or kaolin

1. Antigen must be foreignness to immune system:

. What substances are foreignness to immune system ?

According to Burnnets clone selection theory ,

foreignness ( non-self) means substances that never contact with lymphocytes during embryo period.

What kinds of substances can be foreignness to immune system? (1) Heterogeneous substances Various pathogens, xenoantigeneic tissues

(2) Allogeneic substance

grafted allogeneic tissues or organs (3)Autoantigenic components that never contact with lymphocytes during embryo period

2. Two properties of immunogen

1. Immunogenicity
An ability of antigen which can stimulate the body to evoke a specific immune response. 2. Immunoreactivity An ability of antigen which can combine with

corresponding Ab or sensitized lymphocyte

III. Specificity and cross reaction of antigen Specificity is a cardinal feature of the adaptive immune
system

Specificity is referred to that immune responses are

directed toward and able to distinguish between distinct antigen or small parts of macromolecular antigens. This fine specificity is attributed to lymphocyte antigen receptors that may bind to one molecule but not to another with only minor structural differences from the first

Specificity of Ag

Ab1

Ab2

Ab3

 Specificity exists in both immunogenecity and immunoreactivity Specificity is the basis of immunologic diagnosis and immunologic therapy as well as basic feature of adaptive immunity

 Species specificity:
Tissues of all individual in a species contain species specific antigen Cross reactivity is seen between antigen from related species.

Isopsecificity :
Isoantigen are antigen found in some but not in all members of species. On the basis of isoantigens a species may be divided into different groups Eg. Blood group antigen

 Autospecificity :
A number of tissue antigen may act as a auto or self antigen. Eg cellular nucleic acid, corneal component, thyroglobulin.

Organ specificity:
Some organ of different species share the same antigens are k/a organ specific antigen eg. Brain, spinal cord of one species share specificity with another species

 Heterophile (heterogenetic) specificity:

The same or closely related antigen may sometime occure in different biological species, classes and kingdom are k/a heterophile antigen.

Degradability
Antigens that are easily phagocytosed are generally more immunogenic. This is because for most antigens (T-dependant antigens) the development of an immune response requires that the antigen be phagocytosed, processed and presented to helper T cells by an antigen presenting cell (APC).

 Solubility and susceptibility:

Insoluble do not have effect of tissue enzymes non antigenic Soluble easily metabolized, & susceptible to tissue enzyme antigen

Chemical nature & complexity:

Protein & polysaccharides are more antigenic than lipids and nucleic acid Antigenicity of lipids and nucleic acid may be enhanced by combined with protein. Certain degree of structural complexity is require for antigenicity.

1. Antigen determinants (epitope)

(1) The portion of antigen molecules which can be specifically recognized by antibody or antigenic receptor of lymphocytes.

Protein antigen----5-15 amino acid residues Polysaccharide antigen----5-7 polysaccharide residues

Three dimension figure of Angiotensin(Ag II binding to antibody

Chicken lysozyme bound to an antibody

(2) A change of antigenic determinant (characteristics,

number and conformation) can influence the

specificity of Ag.

Antigen determinant is the sites of Ag combining with Ab

2. Antigenic valence
Total number of determinants which can be bound by

antibody or antigenic receptor of lymphocytes is

called antigenic valence. Most natural antigens are polyvalence antigen.

3. Classification of antigenic determinant

(1)According to the structure of Ag determinants Conformational determinants Sequential (or linear) determinants

Conformational determinants
Conformational determinants are formed by amino acid residues that arent in a sequence but become spatially juxtaposed in the folded protein.

Sequential (or linear) determinants

Epitopes formed by several adjacent amino acid residues are called linear determinants.

(2)According to types of cells recognizing antigenic determinants T cell determinants (T cell epitopes)
B cell determinants (B cell epitopes)

Difference between T cell epitope and B cell epitope

T cell epitope Receptor TCR Nature short peptide Size 8-17 amino acid residues B cell epitope

BCR proteins, polysaccharides 5-15 amino acid residues or 5-7 monosaccharides Types linear epitope conformational epitope or linear epitope Position any position in antigen mostly exist on the surface of antigen

Antibodies
Proteins that recognize and bind to a particular antigen with very high specificity. Made in response to exposure to the antigen. One virus or microbe may have several antigenic determinant sites, to which different antibodies may bind. Each antibody has at least two identical sites that bind antigen: Antigen binding sites. Valence of an antibody: Number of antigen binding sites. Most are bivalent. Belong to a group of serum proteins called immunoglobulins (Igs).

Antibody Structure
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 Ys 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

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.

How Do B Cells Produce Antibodies?

B cells develop from stem cells in the bone marrow of adults (liver of fetuses). After maturation B cells migrate to lymphoid organs (lymph node or spleen). Clonal Selection: When a B cell encounters an antigen it recognizes, it is stimulated and divides into many clones called plasma cells, which actively secrete antibodies.

Each B cell produces antibodies that will recognize only one antigenic determinant.

Clonal Selection of B Cells is Caused by Antigenic Stimulation

Humoral Immunity (Continued) Apoptosis

Programmed cell death (Falling away). Human body makes 100 million lymphocytes every day. If an equivalent number doesnt 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.

Humoral Immunity (Continued) Clonal Selection

Clonal Selection: B cells (and T cells) that encounter stimulating antigen will proliferate into a large group of cells. Why dont we produce antibodies against our own antigens? We have developed tolerance to them. Clonal Deletion: B and T cells that react against self antigens appear to be destroyed during fetal development. Process is poorly understood.

Consequences of Antigen-Antibody Binding

Antigen-Antibody Complex: Formed when an antibody binds to an antigen it recognizes. Affinity: A measure of binding strength. 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.

Consequences of Antibody Binding

Humoral Immunity (Continued)

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

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.

Immunological Memory (Continued)

Secondary Response: Subsequent exposure to the same antigen displays a faster and more intense antibody response. Increased antibody response is due to the existence of memory cells, which rapidly produce plasma cells upon antigen stimulation.

Antibody Response After Exposure to Antigen