IMMUNOGENS AND ANTIGENS - DELFIN, RMT

Immunogen: Any agent capable of inducing an immune response

Antigen: Any agent capable of interacting specifically to components of the immune response
such as lymphocytes and antibodies.
Properties of Immunogen
Immunogenecity: Inherent ability of a substance (immunogen) to induce as specific
response resulting in the formation of antibodies or immune lymphocytes:
Antigenecity: Property of a substance (antigen) to react specifically with the antibody.
Immunogenic substances are always antigenic whereas antigens are not necessary
immunogenic.
Antigens/ Immunogens are found on

RBC surface (Group A and B)

WBC surface(Human Leukocyte antigens)
Platelets
Tissue cells
Semen
Saliva

Antigens/Immunogens can be introduced by:

Blood transfusion
Tissue grafting and organ transplantation
Transplacental leakage
Ingestion

Antigenic Determinant (Epitope)

The portion that binds specifically with the binding site of an antibody (paratope) or to a
lymphocyte receptor.

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Antigenic Determinant (Epitopes)

Consists of 4-6 amino acids or 5-7 monosaccharides in length.

Aromatic amino acids (as tyrosine) provide more immunogenecity that non-aromatic
amines
Polysaccharides are better immunogens than oligosaccahrides because of their
complexity.
Hydrophobic molecules are more immunogenic than hydrophobic molecules.

Configurations of epitopes

Linear: found within the amino acid sequence of the molecule.

Conformational: formed by amino acids that appear on the same area of the protein but
not adjacent to the peptide chain.

Classes of Immunogens:
1. Thymic dependent antigens:

Require the help of T cells for the formation of antibody.

They activate B cells with the help of T cells
Most immunogens are thymic dependent

2. Thymic independent antigens:

Stimulate antibody without interacting with T cells

Response to these antigens is of the IgM class with little or no immunologic memory
generated.
T independent antigens are usually carbohydrates which are composed of
monotonously repeating epitopes.

T-Independent Antigens

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Epitopes, T-Independent Antigen

According to epitopes:

Unideterminant, univalent antigen:

Unideterminant, multivalent antigen:

There is only one kind of epitope but many such epitopes on each molecule such as
many polysaccharides and homopolymers

Multideterminant, univalent antigen:

There is only 1 epitope on the molecule like hapten

There are many different kinds of epitopes but only one of each kind such as protein

Multideterminant, multivalent antigen:

There are many kinds of determinants and many of each kind such as polymerized
proteins

Hapten and Carrier

Haptens:
Low molecular weight compounds which by itself cannot elicit the formation of antibodies but
can combine with an antibody
Also called incomplete or partial antigens
Has the chemical configurations which gives the antigen its antibody provoking activity
Carriers

Usually proteins which give the hapten the required size

Combines with a hapten so that it can elicit formation of an antibody
The hapten temporarily loses it immunogenecity when detached from a carrier and restored if it
combines with the carrier.

Requirements of Immunogenecity

The degree of immunogenecity of a molecule is influenced by several factors. The relationship

is expressed as:

Immunogenecity = (Foreigness) (chemical complexity) (molecular size)

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1. Foreigness
An antigen must be foreign or non self to the host to which it is administered.
o Autologous Antigens: found within the same individual
Autograft
o Syngeneic antigens: found in genetically identical individuals
Syngeneic graft or isograft
o Allogenic or Homologous antigens: Found between different individuals with the
same specie
Homograft/allogragft
o Xenogeneic or heterologous antigens: Found between different individuals of
different species
Xenograft
o Sequestered Antigens: Antigens not exposed to antibody producing cells and become
immunogenic when exposed to antibody forming tissues.
o Heterogenetic/Heterophilic antigens: Occur in unrelated animal and plant species
2. Chemical Complexity
Proteins:
o Majority of immunogens
o Strongest immunogens because of the diversity of amino acids which imparts epitopes of
different specificities to the molecules
o The total immune response will be the sum of all the antibodies produced by the epitopes
Glycoproteins
o Immunogens
o Best illustrates by RBC blood group antigens
o HLA antigens found on the surface of nucleated body cells comprising both solid tissue
and most circulating blood cells.
Polysaccharides:
o Most are haptens
o Easily degraded when injected
Pure polysaccharides substance
Pneumococcal capsule which are responsible for the protective immune
response to pneumococcus
Lipopolysaccharide substances (endtotoxins)
Nucleic Acids:
o Non immunogenic because of relative simplicity, molecular flexibility and rapid
degradation
o Nucleoproteins are stronger immunogens because the nucleic acids are coupled to
proteins.
Lipids
o Non immunogenic
Relatively simple, lack structural stability
3. Molecular Size
As a general rule, molecules below 5,000 daltons are non-immunogenic.
Reasonable immune responses will be induced by molecules like
o serum albumin (40,000 daltons)
o Gamma globulin (160,000 dlatons)
o Hemocyanin (1,000,000)
Size is important since the number of epitopes increases proportionately with the size or
protein.
Larger molecules from antibodies move rapidly since they are processes and phagocytized by a
macrophage.
Soluble antigen is difficult or impossible to phagocytized and therefore is not immunogenic at
times.

Genetic Composition
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Major Histocompatibility Complex

o Exerts genetic influence of an individuals ability to respond to an immunogen.
o It has been found that if a particular MHC antigen is not present for an immunogen, there
is no specific response.

Route Dosage and Timing

Intravenous and intraperitoneal routes are effective

Intradermal route offers a stronger stimulus than the subcutaneous or intramuscular route,
Generally, the smaller the dose, the less likely an immune response

Cross Reactivity

Denotes a situation in which two or more compounds that may have various degree of
dissimilarity, share antigenic determinants/epitope and would therefore react within immune
components induced against any one of the compounds.
A toxoid is a modified form of toxin may have one more antigenic determinants in common with
the native toxin
Immunization with a toxoid leads to an immune response capable of reacting only with the
toxoid but also with the native toxin
Cross reactivity is illustrated by:
o Treponema pallidum and cardiolipin
o Epstein-Barr Virus and sheep red blood cells
o Ricketssia and Proteus

Immunologic Adjuvant

Adjuvant: Substance which when mixed with an Immunogen enhances the immune response
against the Immunogen

Types of Adjuvant
1. CFA (Complete Freunds Adjuvant)

Consist of water-in-oil emulsion and killed Mycobacterium tuberculosis or butyricum

The antigen in in the water phase.
o Other examples:
o BCG
o Corynebacterium parvum
o Bordetella pertusis
These types of adjuvants release the antigen slowly but continously and stimulate certain
subpopulation of T lymphocytes and macrophages.

2. Toxins and polysaccharide

a. LPS (lipopolysaccharide or endotoxin)
b. Synthetic MDP (muramyldipeptide)
LPS enhances antibody response by stimujlating subpopulation of B cells while MDP stimulates Tcells
3. Alum precipitate

Consist of alumunum hydroxide on which antigen is absorb

Has a slight irritant effect which enhances ingestion and processing of antigen by macrophages.

Hapten is different from adjuvant

Hapten becomes immunogenic when conjugated covalently to a carrier but will not become
immunogenic when conjugated with an adjuvant.
An adjuvant enhances immune response to Immunogen but does not confer immunogenecity to
Haptens

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Second Line of Defense

1.
2.
3.
4.

Phagocytosis
Inflammation
Complement System
Acute phase reactants

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1. Phagocytosis
(engulfment and destruction) of microorganism and damaged cells
literally means eating cell process
Phagocytosis MEMORIZE (ICED)
o Initiation is caused by damage to the tissues, either by trauma or as a result of
microbial multiplication.
o Chemotaxis, attraction of leukocytes or other cells by chemicals.
Opsonization - Opsonization coating a pathogen by substances so as to
enhance phagocytosis.
Adherence - firm contact between phagocyte and microorganism.
o Engulfment into cytoplasm and enclosed in a vacuole.
o Digestion enzymatic contents in vacuole destroy the microorganism.
INITIATION is caused by damage to the tissues, either by trauma or as a result of
microbial multiplication.
activated phagocyte has increased surface receptors that allow for adherence
(adhesive factor)
o CR3

o Lamin receptor
o Leucyformyl-methionyl
phenylalanine
CHEMOTAXIS attraction of leukocytes or other cells by chemicals. ( chemotaxin)
Chemotaxin
o Interleukin 1 - released by macrophage
o Histamine - released by basophils, tissue mast
cells
o Complement (C5a)
2 kinds
o Positive movement towards the stimulate
o Negative movement away from the stimulate
Opsonization - a process of coating a pathogen by substances so as to enhance
phagocytosis.
o to prepare for eating.
o Opsonins are serum proteins that attach to a foreign substance and help
prepare it for phagocytosis
o Opsonin:
antibody
Complement component
C- reactive protein
Adherence - firm contact between phagocyte and microorganism.

ENGULFMENT
achieved through amoeboid motion

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 Phagosome (vacuole) - contained the engulfed materials

Phagolysosome - fusion of phagosome and lysosome
DIGESTION
o enzymatic contents in vacuole destroy the microorganism.
Number of killing mechanisms operating in the vacuoles of phagocytic cells.
One of the major mechanisms involves hydrogen peroxide which, acting along with
an intracellular enzyme, is rapidly lethal to many bacteria.
Destruction of antigen
1. Formation of the phagolysosome leading to the release of lysosomal contents including
defensins,
lactoferrin
lysozyme.
2. Nitric oxide produce by macrophage which is toxic to microorganism
3. Activation of the NADPH oxidase, which is present in the phagosome membrane, leading to the
production of reactive oxygen intermediates, superoxide anion ,H2O2 and hyrogen radicals
which are cytotoxic for microorganism.
Second Line of Defense
Phagocytosis
Inflammation
Complement System
Acute phase reactant

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INFLAMMATION
A nonspecific exaggerated physiological response by phagocytic cells to infection/injury.

Rubor (redness) is caused by increased circulation and vasodilation in the injured tissues;
Calor (warmth) is the heat given off by the increased flow of blood;
Tumor (swelling) is caused by increased extracelluar fluid accumulating in the tissues
Dolor (pain) is caused by the stimulation of nerve endings from the pressure of swelling or
by chemical mediators
Functio laesa (loss of function)

STAGES

Vascular response
Cellular response

Vascular response

dilation of blood vessels to increase blood flow in the area of injury

hyperemia
redness/ heat
increase capillary permeability
plasma leakage to tissue

Cellular response

Diapedesis squeezing out of WBC from endothelial space

Neutrophils

first to migrate
short lived
acute inflammation

Monocyte/ Macrophage

2nd to migrate
long lived
chronic inflammation
act as APC (antigen presenting cell)
APC (antigen presenting cell)
o release interleukin 1 (IL1)
o Effects of IL1
Act in hypothalamus (FEVER)
Increase acute phase reactants

ACUTE PHASE REACTANTS

REPAIR AND RESOLUTION

Initiated by fibroblast proliferation

affected area may be totally repaired

injury may lead to formation of abscess
granuloma may be formed

Antimicrobial substances

Complement
Properdin
Interferon
Tumor necrosis factor
Betalysin

1. Interferon - glycoproteins that has virus- nonspecific antiviral activity

Type 1 non- immune, produced primarily in the initial innate response to viral infection
o

alpha IFN
aka leukocyte interferon
produce by virus induced leukocyte culture (NULL lymphocyte)
beta IFN
aka fibroblast & epithelial/ fibroepithelial IFN
produced by fibroblast cells

Type 2 immune, primarily produced as a component of the specific immune response to viral
and other pathogens
Gamma IFN
aka immune IFN
produced by immunologically stimulated lymphocyte
Tumor Necrotic factor
cytotoxic against tumor cells and virally infected cells
TNF alpha
o aka cachetin
o produced by macrophage
TNF beta
o aka lymphotoxin
o produced by CD4 and CD8 cells
Complement - complex series of more than 30 soluble and cell-bound proteins that interact in
a very specific way to enhance host defense mechanisms against foreign cells.
Function
o mediate inflammation
o activate immune system
o opsonization
o cell lysis
Properdin - serum protein that exerts bactericidal and viricidal effect in the presence of C3 and
Mg
Betalysin
released by platelets during coagulation
substance with bactericidal activity found in serum (not in plasma).
o

2.

3.

4.
5.

Interleukin - cytokines (chemical messengers) produced by leukocytes that affect the inflammatory
process through an increase in soluble factors or cells
Cellular versus Humoral Immunity

Cellular - foreign substances were removed by specialized cells in a process known as

phagocytosis.
Humoral - substances in the blood provided protection from microorganisms, humoral
immunity

ANTIBODIES - specific proteins referred to as immunoglobulins

Many are isolated in the gamma globulin fraction of protein (electrophoresis separation)
Found in the plasma and many body fluids (eg. Tears, saliva, colostrums)
Immunoglobulin (Ig)
Function
o 1 - body defense: combine w/ antigen, enough to neutralize bacterial toxins or some
viruses

Immunoglobulins
Classes differ from characteristics such as MW and sedimentation coefficients
1. IgM (SC: , of 19)
10 % of Ig pool
Largely confined to intravascular pool and blood due to its large size
Produced early in immune response
Effective in Agglutination & Cytolytic reactions
5 individual HEAVY chains (65,000 Da, whole: 900,000 Da)
Decreased in 1 (genetically determined) Ig disorders as well as 2 Ig deficiencies
Increased in
o Infectious diseases, such as subacute bacterial endocarditis, infectious
mononucleosis, leprosy, trypanosomiasis, malaria, and actinomycosis
o Collagen disorders, such as scleroderma
o Hematologic disorders, such as polyclonal gammopathies, monocytic leukemia,
and monoclonal gammopathies(e.g., Waldenstrms macroglobulinemia)
PENTAMER
2. IgG (SC: 7S)
Major Ig in normal serum
Diffuses more readily than other Igs into extravascular spaces and neutralizes toxins or
binds to microorganisms in extravascular spaces
Only Ig that cross placenta except IgG2
MW: 150,000 Da
Cord blood and CSF
Decrease in 1 (genetically) or 2 (acquired) Ig deficiencies
Significant Increase in
o Infectious diseases, such as hepatitis, rubella, and infectious mononucleosis
o Collagen disorders, such as rheumatoid arthritis and systemic lupus
erythematosus
o Hematologic disorders, such as polyclonal gammopathies, monoclonal
gammopathies, monocytic leukemia, and Hodgkins disease
MONOMER
3. IgA

4. IgD

15&-20% total circulatory Ig pool

Predominant Ig in secretion of tears, saliva, colostrums, milk and intestinal fluids
Synthesized largely by plasma cells on BODY SURFACES
May pass directly into intestinal lumen or diffuse into blood circulation
Transported through intestinal epithelial cells (hepatocytes) = binds to glycoprotein called
SECRETORY COMPONENT
o Protects Ig from digestion by GI proteolytic enzymes
o Comple IgA - critical in protecting body surfaces against invading miciroorganisms
o Presence of seromucous secretion (tears, saliva, nasal fluids, colostrums)
Monomer and Dimer
Decrease in 1 (genetically) or 2 Ig deficiencies
Significant increase in serum
o Infectious diseases, such as tuberculosis and actinomycosis
o Collagen disorders, such as rheumatoid arthritis
o Hematologic disorders, such as polyclonal gammopathies, monocytic leukemia, and
monoclonal gammopathy (e.g., IgA myeloma)
o Liver disease, such as Laennecs cirrhosis and chronic active hepatitis

Low concentration in plasma, <1% total Ig pool

Extremely susceptible to proteolysis
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5. IgE

Primary a cell membrane found on the SURFACE of B lymphocytes in association w/ IgM

MONOMER
Trace plasma protein found in blood plasma of unparasitized individuals
MW: 188,000 Da
Mediates some types of hypersensitivity (allergic reactions, allergies, and anaphylaxis
Binds strongly to receptor on mast cells and basophils
w/ antigen, mediates release of histamines and heparin
MONOMER

Antibody Structure

Homology unit or Domain

o 12 Domains
o 2 Heavy Chains and 2 Light Chains linked through cysteine residues by disulfide bonds
o Variable Region N-terminal end (heterogeneity); antigen-binding portion
o Constant Region constant amino acid sequences
Globular Regions
o 2 Fab and Fc region linked by Hinge Region
o 2 Fab fragments retain ability to bind antigen
o Fc portion homogenous, crystallizable

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