Antigen

Subs that stimulate antibody formation

Has the ability to bind with antibody

Antigenic determinants or Epitopes

Part of an antigen(membrane or molecular) Structures recognized as foreign by the immune

system Reacts specifically with an antibody or Tlymphocyte receptor

Antigen
Histocompatibility Antigens
Autoantigens

Blood group Antigens

Histocompatibility Antigens
Constitute the MHC(Major

Histocompatibility Complex) or HLA( Human Leukocyte Antigen) Nucleated cells such as leukocytes and tissues possesses many cell

Chemical Nature of Antigens

Proteins
Excellent Antigens High MW and structural complexty

Lipids
Poor antigens Simplicity and lacks structural stability

Nucleic Acids
Poor antigens Simplicity, molecular flexibility and rapid degradation

Carbohydrates
Poor antigens Too small

Physical Nature of Antigens

Foreigness
Degradability Molecular Weight The higher the MW, the better

Hapten-tiny molecules that can bind to a larger molecule and behave as antigens

Structural Stability

Complexity The greater the complexity, the greater its effectiveness

Antibodies
Immunoglobulin
Found in blood plasma and in many body fluids Primary function is to combine with antigen Maybe enough to neutralize bacterial toxin or some viruses

Secondary interaction with another effector agent is usually required to dispose of larger antigen

Immunoglobulin Classes
Ig M
Ig G Ig A

Ig D
Ig E

Ig M
10% of the Ig pool
Largely confined to the intravascular pool because of its

large size Produced early in an immune response Function effectively in agglutination ad cytolytic reactions Normal values:

males 60-250 mg/dL Females 70-280 mg/dL

Ig M
Increased in: Infectious diseases such as subacute endocarditis, infectious mononucleosis, leprosy, trypanosomiasis, malaria and actinomycosis Collagen disorders such as scleroderma Hematological disorders such as polyconal gammopathies, monocytic leukemia and monocolonal gammopathies

Ig G
Major immunoglobulin found in normal serum
Diffuses into extravascular spaces where it can bind to

toxins and microorganism Can cross placenta When Ig G complexes are formed, complement can be activated Normal values:

Adult 800-1800 mg/dL Infants(3-4 months) 350-400 mg/dL 1st year 700-800 mg/dL

 Increased in: Infectious diseases such as hepatitis, rubella, and infectious mononucleosis Collagen disorders such as rheumatoid arthritis and systemic lupus erythematosus

Ig A
15-20%
Predominant Ig in secretions such as tears, saliva,

colostrum, milk and intestinal fluids Synthesized largely by plasma cells located on body surfaces If produced by cells in the intestinal wall, it may pass directly into the intestinal lumen or diffuse into the blood circulation As Ig A is transported through intestinal epithelial cells or hepatocytes, it binds to a glycoprotein called secretory component

 Secretory component Protects Ig A from digestion by gastrointestinal proteolytic enzymes Forms complex molecule called secretory Ig A
Secretory Ig A Protects body surfaces because of its presence in

seromucous secretions

Ig D
Accounting less than 1% of the total Ig pool
Susceptible to proteolysis and is primarily a cell

membrane

Ig E
Trace plasma protein found in the blood plasma protein

found in the plasma of unparasitized individuals Mediates some types of hyper sensitivity(allergic) reactions, allergies, and anaphylaxis and is generally responsible for an individuals immunity to invading parasites It binds strongly to a receptor on mast cells and basophils and together with antigen, mediates the release of histamines and heparin from these cells

 Increased in: Infectious disease such as tuberculosis and actinomycosis Collagen disorders such as rheumatoid arthritis Hematologic disorders such as Liver disease such as Laennecs cirrhosis and chronic active hepatitis

Ig M % in blood 10% confne

Ig G 70-75%

Ig A 15-20%

Ig D <1%

Ig E

Antibody Structure
Polypeptide with 3 dimensional

configuration Has 2 regions:

One region of the molecule involves

binding to antigen(Fab region) A diff region mediates binding of the Ig to host tissues including cells of the immune system and the first component (C1q) of the classic complement system (Fc portion)

TYPICAL IMMUNOGLOBULIN MOLECULE

Domain Basic unit of an antibody structure A typical molecule has 12 domains arranged in two heavy (H) and two light(L) chains linked through cystein residues by disulfide bonds so that domains lie in pairs

 Variable region(Fab) Antibody-binding region Constant region(Fc)

Composed of relatively constant amino acid sequences

Papain
Splits a monomer into three approximately equal-size

fragments:

2 Fab 1 Fc

Pepsin
Splits monomer differently: The Fc fragment is split into tiny fragments and is completely destroyed The 2 Fab fragment remain joined to produce a fragment called F(ab)2 which possess two antigen binding site

Fd fragment
Fd fragment
contains a light chain and half of a heavy chain

Structure of Other Immunoglobulins

IMMUNOGLOBULIN VARIANTS
Antigenic determinant/Epitope
Specific chemical determinant group or molecular

configuration against which immune response is directed

3 principal categories :
Isotype Determinants -dominant type found on the Ig

of all animals of a specie Allotype Determinants -may be produced by injecting the Ig of one animal into another member of the same species Idiotype Determinants -result of the unique structures on light and heavy chains

ANTIBODY SYNTHESIS

Induced when the hosts lymphocytes come in contact with a foreign antigenic substance that binds to its receptor

Clonal selection

Activation and proliferation Clonal expansion of lymphocytes in response to infection necessary for an effective immune response Requires 3-5 days to produce sufficient number of clones and to differentiate into antibody-producing cells

Primary Antibody Response

1. Lag phase no antibody is detectable
antibody titer increases logarithmically antibody titer stabilizes antibody is catabolized 2. Log phase 3. Plateau phase 4. Decline phase

Secondary(Anamnestic) Response

Subsequent exposure to the same antigenic stimulus Differs from a primary response as follows:
has a shorter lag phases, longer plateau and a more gradual decline Ig G class is the predominant type formed antibody levels attain a higher titer

1. Time 2. Type of Antibody 3. Antibody titer

FUNCTION OF ANTIBODIES

Principal function is to bind to Antigen May also exhibit secondary effector functions and behave as antigens Significant secondary functions of antibodies are complement fixation and placental transfer
*Placental transfer is important in the etiology of Hemolytic Disease of the Newborn and Conferring passive immunity to the newborn during the first few months of life

Comparison of Properties of Immunoglobulins

Ig M Ig G
Complement Fixation

Ig A

Ig D
No No

Ig E
No No

3+ No

0-2+ No Yes No

Placental Transfer

ANTIGEN-ANTIBODY INTERACTION SPECIFICITY AND CROSS-REACTIVITY

Specificity

Ability of a particular antibody to combine with a particular antigen Exist when the binding sites of antibodies directed against determinants of one antigen are not complementary to determinants of another dissimilar antigen

 Combining site

Portion of the Fab molecule Cleft formed largely by the hypervariable regions of heavy and light chains **The closer the fit between this site and the antigenic determinant, the stronger are the noncovalent forces and the higher is the affinity between the antigen and the antibody **When more than one combining site interacts with the same antigen, the bond has greatly increased strength

 Cross-reactivity

When a proportion of the antibodies directed against onetype of antigen will also react with the other type of antigen *Heterophile antibodies

Antibody Affinity
Affinity

Initial force of attraction that exist between a single Fab site on an antibody molecule and a single epitope or determinant site on the corresponding antigen Antigen is univalent and is usually a hapten

Antibody Avidity
Avidity

Functional combining strength of an antibody to its antigen when a multivalent antigen combines with more than one of the antibodys combining sites, the strength of the bonding is significantly increased

Immune Complexes

Noncovalent combination of antigen with its respective specific antibody

Small Large

soluble precipitating

Once formed in the circulation, the immune complex is usually removed by phagocytic cells through the interaction of the Fc portion of the antibody with complement and cell surface receptors May be viewed as major host defense against the invasion of foreign antigens

 *If the immune complex becomes soluble and escapes

phagocytosis , it may cause the ff:

deposited in endothelial or vascular structures(causing inflammatory damages) Deposited in organs uch as the kidneys Inhibits useful immunity

MOLECULAR BASIS OF ANTIGEN-ANTIBODY REACTIONS

Noncovalent bond
Weak compared to covalent bonds, but formation of

multiple noncovalent bonds produces a considerable total binding energy

Four types of noncovalent bonds:

1. Hydrophobic bonds

Major bonds Hydrogen bridges between appropriate atoms Interaction between electron clouds and hydrophobic bonds Attraction of oppositely charged amino acids located on the side chains of two amino acid residues

2. Hydrogen bonds

3. Van der Waals Forces

4. Electrostatic Forces

Detection of Antigen-Antibody reactions

Agglutination

Process in which particulate antigen aggregate to form larger complexes in the presence of a specific antibody Widely used in immunology to detect and measure the consequences of antigen-antibody interaction

 Precipitation reactions Combine soluble antigen with soluble antibody to produce insoluble complexes that are visible Hemolysis Testing Involves the reaction of antigen and antibody with a cellular indicator Enzyme-linked Immunosorbent Assay(ELISA) Measures immune complexes formed in an in vitro system