Immunity

It is the ability of host body to prevent or overcome invasion by virulent microorganism. It is

known as resistance or immunity.

Types of immunity:

A. Natural immunity

B. Acquired immunity/Adaptive Immunity

A. Natural immunity
It is also called non-specific immunity because it exists in all humans and is present from earliest
time of life. It provides defense against infections by a number of chemical and mechanical
barriers. These natural barriers include skin, mucous membrane, secretions, and component of
blood, enzymes and often body fluids. These barriers form First line of defense against infections
and diseases.

Susceptibility:

Lack of such natural resistance is called susceptibility.

Components of innate immunity :

1.Skin layer:

It provides a protective covering to all body tissues.

2.Mucous membrane:

It secretes sticky mucous, that trapped the air born particles and sweep them out by ciliary
movement.

3.Acidity of stomach and vagina:

Acidic pH is toxic to most microorganisms.

4.Bile:

Bile is inhibitory to most microorganisms.

5.Lysozymes:

It is present in saliva and digest cell wall of gram positive bacteria.

6.Microbial flora:
The normal microbiota prevent pathogens from colonizing the host by competing with them for
nutrients (competitive exclusion)
Second line of defense:

Once microorganism succeed in passing first line of defense, they enter the deeper tissues and
are attached by specific cells of the body, which may ingest or destroy them. These cells are
called phagocytes and they form second line defense.

Types:

There are two types

a. Free phagocytes.

b. Fixed phagocytes.

a)Free phagocytes:

It includes neutrophils that are present in blood stream.

b)Fixed phagocytes:

These are the macrophages of reticular endothelial system. They are present in tissues.

Phagocytosis:

Greek words meaning eat and cell is the ingestion of a microorganism or other substances (such
as debris) by a cell. It is the process by which phagocytes engulf the foreign particles in the form
of phagosomes.It involves following steps.

1. Adequacy of blood flow.

2. Leukocytes adhesion to capillary walls and passing through it.
3. Chemotaxis is the chemical attraction between parasites and phagocytes mediated by a
substance released by parasite.
4. Amoeboid engulfment of parasite in the form of phagosome.
5. Fusion of phagosomes with lysozymes and final digestion.
6. Removal of undigested particles outside the phagocytes.
II. Opsonization(Enhanced phagocytosis):
Sometimes, the pathogens repels the phagocytes (-ve chemotaxis) in such situation the
component of complement systemopsonin bind the parasite to phagocyte it. This enhanced
phagocytosis called opsonization.

Opsonin:

They are serum protein that increases the susceptibility of parasite to phagocytosis.

III. Complement system:

It is a group of complex system of 30 serum proteins and other factors produced in liver and are
present in normal serum of all vertebrates .They play a major role in animal defense.
Inflammation

Damage to the body's tissues triggers a local defensive response called inflammation, another
component of the second line of defense. The damage can be caused by microbial infection
( lipopolysaccharide of microorganism may induce inflammatory response), physical agents
(such as heat, radiant energy, electricity, or sharp objects), or chemical agents (acids, bases, and
gases).

Inflammation has the following functions:

(1) To destroy the injurious agent, if possible, and to remove it and its by-products from the
Body. Influx of phagocytes from capillaries to tissues is facilitated by increased permeability of
capillaries.

(2) If destruction is not possible, to limit the effects on the body by confining or walling off the
injuriousagent and its by-products

(3) To repair or replace tissuedamaged by the injurious agent or its by-products

Fever

An abnormally high body temperature, a third componentof the second line of defense .The most
frequentcause of fever is infection from bacteria (and their toxins) orviruses.
Up to a certain point, fever is considered a defense against disease. Interleukin-I helps step up the
production o f T cells. High body temperature intensifies the effect of antiviral interferon and
increases production of transferrins that decrease the iron available to microbes. Also, because
the high temperature speeds up the body's reactions, it may help body tissues repair themselves
more quickly. However death results if temperature rises above 44-46C.

Interferones

Interferons (IFNs) are a class of similar antiviral proteins produced by certain animal cells, such
as lymphocytes and macrophages, after viral stimulation. One of the principal functions of
interferons is to interfere with viral multiplication.
 IMMUNITY

NATURAL IMMUNITY(NON-SPECIFIC) ACQUIRED IMMUNITY(SPECIFIC)

ACTIVE PASSIVE
IMMUNITY IMMUNITY

NATURAL ARTIFICIAL NATURAL ARTIFICIAL

ACQUIRED ACQUIRED ACQUIRED ACQUIRED
IMMUNITY IMMUNITY IMMUNITY IMMUNITY

Acquired immunity(specific defense):

It involves the formation of antibodies(third line of defense) as a result of stimulation by foreign

particles (Antigen). Acquired immunity is developed during individuals life time.

Adaptive immunity displays four characteristic attributes:

■ Antigenic specificity

The antigenic specificity of the immune system permits it to distinguish subtle differences
among antigens. Antibodies can distinguish between two protein molecules that differ in only a
single amino acid.

■ Diversity

The immune system is capable of generating tremendous diversity in its recognition molecules,
allowing it to recognize billions of unique structures on foreign antigens

■ Immunologic memory

Once the immune system has recognized and responded to an antigen, it exhibits immunologic
memory; that is, a second encounter with the same antigen induces a heightened state of immune
reactivity.

■ Self/nonself recognition

Finally, the immune system normally responds only to foreign antigens, indicating that it is
capable of self/nonself recognition
It is of two types.

A. Active immunity:
It is the immunity that develops from exposure to antigen that lead to production
of antibodies. Exposure to antigen may be intentional or unintentional.

Types of active immunity

It is of two types

a)Natural acquired Active immunity:

It results due to unintentional exposure of antigen e.g. Immunity that develops
after an infection such as mumps and measles etc.

b)Artificial acquired Active immunity:

It develops as a result of intentional exposure to antigen e.g.vaccine

administration.

B. Passive immunity:
It is the immunity that develops from infusion of antibodies from outside source.

Types of passive immunity:

It is of two types.

a)Artificial acquired Passive immunity:

It is due to intentional injection of antibodies rich serum into circulation, e.g. Hepatitis
A ,antiserum, chicken pox antiserum .

b)Natural acquired Passive immunity(congenital immunity):

This immunity develops when antibodies pass into fetal circulation from mother blood through
placenta and also form colostrums.
Difference between active and passive immunity

Active immunity Passive immunity

1 Active immunity is the 1. immunity develops from anti-bodies

. immunity that develops from
exposure to antigen that lead to
production.

2 It develop slowly not become 2. It is the fast process because we directly

. fully effective even for several introduce anti-bodies. Established
weeks. quickly the time depends upon route of
administration.

3 Once developed it is long 3. It is very short lived because body

. lasting. it after gives protection recognize the antibodies as foreign
for many years even when it substance and gradually eliminate them.
begins to fade a booster dose of Protection is only for 2-3
antigen quickly restore the weeks.exception is, when human anti-
effective anti-bodies level. bodies are given, the rate of elimination
is slower in this case.

4 Active immunity is mainly 4. It is both for prophylaxis therapeutic

. used for prevention of diseases. purpose.
(prophylaxis)

5 Examples: polio vaccine, 5. Examples: Tetanus anti toxin to prevent

tuberculosis vaccine. tetanus after road accident, Diphtheria
anti toxin.
IV. Duality of immune system:

In vertebrates two parallel immune responces have been award namely cellular or cell mediated
immune response and humoral or anti-body mediated immune response to cope with widely
different categories of invaders to defend some classes of organism both humoral orcellular
responsees are needed. Antigens, which are generally very large and complex, are not recognized
in their entirety by lymphocytes. Instead, both B and T lymphocytes recognize discrete sites on
the antigen called antigenic determinants, or epitopes

DUALITY OF IMMUNE SYSTEM

HUMORAL OR ANTI-BODY
CELL MEDIATED IMMUNITY
MEDIATED IMMUNITY

A. Cell mediated immune response:

This response is not due to antibodies but it is due to sensitized thymus derived lymphocytes
that is T-lymphocytes’-cells develop from stem cells in the bone marrow. Mature in thymus
gland where they are ready to in counter antigen.The cell-mediated branch (T cells) recognizes
protein epitopes displayed together with MHC molecules on self-cells, including altered self-
cells such as virus-infected self-cells and cancerous cells. T-lymphocytes recognize invading
organism as foreign and initiate chain reaction as follows.

1. Cytotoxic destruction of invading cells.

2. Activation of phagocytic macrophages. This system is primarily responder to
transplanted tissues such as foreign skin graft. It response is to reject foreign
tissue. It is also an important factor in our defense against cancer.
 3. The cell mediated immune response is most effective against bacteria, viruses
located within phagocytic and infected host cells and against fungi, protozoa, and
helminth.

B. Humoral or anti-body mediated immunity:

It involves the production of anti-bodies that act against foreign organism and substances. Cells
called B-cells or B- lymphocytes are responsible for production of anti-bodies. These anti-bodies
are found in extra-cellular fluid such as plasma, lymph and mucous secretion. The humoral
branch (B cells) recognizes an enormous variety of epitopes: those displayed on the surfaces of
bacteria or viral particles, as well as those displayed on soluble proteins, glycoproteins,
polysaccharides, or lipopolysaccharides that have been released from invading pathogens. The
humoral immune response defend primarily against bacterial toxin, bacteria and virus that are
circulating freely in the body fluids. It is also a factor in some reactions against transplanted
tissues.

The lymphatic system

The lymphatic system consists of a fluid called lymph, vessels called lymphatic vessels, a number
of structures and organs containing lymphoid tissue, and red bone marrow, where stem cells
develop into blood cells, including lymphocytes

Lymphocytes:
Lymphocytes are basic cell responsible for both humoral and cellular immunity. Bone marrow
stem cells are ultimate organ of erythrocytes and all leucocytes including lymphocytes
production. Many lymphocytes pass through thymus where they become processed by hormonal
micro environment try to release. These lymphocytes are now called thymus derived or
lymphocytes T-cells. During its maturation within the thymus, the T cell comes to express a
unique antigen-binding molecule, called the T-cell receptor, on its membrane. Unlike membrane-
bound antibodies on B cells, which can recognize antigen alone, T-cell receptors can recognize
only antigen that is bound to cell-membrane proteins called major histocompatibility complex
(MHC) molecules. There are two well-defined subpopulations of T cells: T helper (TH) and T
cytotoxic (TC) cells. T helper and T cytotoxic cells can be distinguished from one another by the
presence of either CD4 or CD8 membrane glycoproteins on their surfaces.T cells displaying CD4
generally function as TH cells, whereas those displaying CD8 generally function as TC cell

By majority of bone marrow derived lymphocytes which do not enter or become processed by
thymus are called B-lymphocytes or B-cells.
Lymphatic system consist of

A. Primary lymphoid system

B. Secondary lymphoid system

C. Primary lymphoid system:

Primary lymphoid organ provide environment for development, differentiation and maturation of
cells. Thymus and bone marrow.

D. Secondary lymphoid system:

Those organ where mature B and T lymphocytes settle and initiate immune response e.g. lymph
nodes,spleen, liver etc

ANTIGEN:
ANTIGEN is derivedfrom two words.ANTI means antibody and GEN means generation.Antigen
are substances which help in generation of antibody.

DEFINITION:
Substances which when inoculated into animal body provoke new response by stimulating the
cells of immune system to produce modified globulin called anti bodies.
PROPERTIES OF ANTIGEN:
Antigen exhibit following properties.

IMMUNOGENECITY:

Antigen should have ability to stimulate immune system.

REACTIVITY:
Antigen should have ability to react with immune components

CHEMICAL NATURE OF ANTIGEN

Most antigens are either protein or large polysaccharide.Lipids and nucleic acids are antigen only
when combined with proteins and polysaccharides

TYPES

There are following types of antigen.

1-Microbial antigen

2-Non microbial antigen

MICROBIAL ANTIGEN:
Antigenic compounds are often comprises of invading microorganism such as
capsules,cellwall,flagella,pilli,and toxins of bacteria,Coats of viruses or surface molecule of
other types of microbes.

NON MICROBIAL ANTIGEN:

They include pollens,egg white ,blood cell surface molecules,Serum proteins from other
individual or species and surface molecules of transplanted tissue and organs.

ANTIGENIC DETERMINENT

Generally antibodies recognize and interact with specific region on antigen called antigenic
determinant or epitope.
Most antigen have a molecular weight of 10000 Dalton or higher.A foreign substance that has a
low molecular weight is often non antigenic unless attached to a corner molecule .These small
molecules are hapten.

e.g. Penicillin is a good example of hapten.

This drug is non antigenic by itself but some people develop an allergic reaction to it.In the
people penicillin combine with serum proteins.The resulting combined molecule initiate immune
response.

ANTIBODIES

Antibodies are proteins that are made in response to antigen and can recognize and bind to that
antigen.Antibodies can therefore help to neutralize or destroy that antigen.Antibodies are highly
specific in recognizing the antigen and antigens such as bacterium or virus may have several
antigenic determinents or epitopes that cause production of different anti bodies.Each antibody
has at least two identical sites that bind to antigenic determinants.These sites are known as
antigen binding sites.

VALENCY:
No. of antigen binding sites is called valency of that antibody.

e.g. Most human antibodies have two binding sites therefore are Bivalent.

ANTIBODY’S STRUCTURE:

Because of bivalent antibody have simple molecular structure,it is called as monomer.A typically
monomer have 4 proteins chains.Two identical light chains[L CHAIN],two identical heavy
chain[H CHAIN].Light and heavy chains refer to their relative molecular weight .These chain
are joined by disulphide links to form a ‘’Y’’shapedstructure.Constant and variable region are
exist in each chain.
VARIABLE REGION :
Variable region structure reflects the nature of antigen for which they are specific.

CONSTANT REGION:
There are 5 major types of constant for 5 major classes of immunoglobulins.Stem of ‘Y’shaped
monomer antibodies called Fc region Fc fragment.So named because in the early days when
antibody structure was being identified,it was afragment that crystallized in cold storage.Fc
region are often important in immunological reactions.
ANTIGEN ANTIBODY REACTIONS

Immunology serves as diagnostic tool, used for the detection of diseases by interaction
ofantibodies with antigens. These reactions include:
1: Agglutination reaction

2: Precipitation reaction

3: Neutralization reaction

1: Agglutination reaction:

Serological test in which antigen molecules are attached to large particles when combined with
patient’s serum containing complementary antibodies result in the visible clumps.

Types:

A: Direct agglutination test

B: Indirect agglutination test

C: Heme agglutination test

Direct agglutination reaction:

Antigens are part of large molecules such as bacterium.

Indirect agglutination reaction:

Antigens or antibodies are absorbed on the surface of latex beads.

Heme agglutination reaction:

Agglutination reaction in which clumping of RBC’s is carried out is called heme agglutination
reaction.

Examples of agglutination reactions:

I: Widal test:

Purpose. It is used for diagnosis of typhoid.

TEST OBSERVATION RESULT

Typhoid bacilli Agglutination Positive

Person serum No agglutination Negative

II: Weil-Fliex Test:

Purpose. It is used fordiagnosis ofRickettsial infection.

 TEST OBSERVATION RESULT

Proteus org. + patient serum Agglutination Serumhas complementary

antibodies so person has
rickettsial infection.

Proteus org. patient serum No agglutination Serum has no complementary

antibodies so person doesn’t
have rickettsial infection.

III: TPA Test: (TreponemaPallidium Agglutination Test).

TEST OBSERVATION RESULT

Living or dead treponema + Agglutination Positive

patient serum.

Living or dead treponema + No agglutination Negative

patient serum.

2: Precipitation Reaction:
Antibody and soluble antigen interacting in aqueous solution form a lattice that eventually
develops into a visible precipitate. Antibodies that aggregate soluble antigens are called
precipitins. Although formation of the soluble Ag-Ab complex occurs within minutes, formation
of the visible precipitate occurs more slowly and often takes a day or two to reach completion.

Precipitation reaction should be used in following ways:

A: Tube precipitation test

B: Slide precipitation test

C: Gel effusion test

Diagnostic application:

It is used for diagnosis of anthrax by Ascoli Test. Although various modifications of the
precipitation reaction were at one time the major types of assay used in immunology, they have
been largely replaced by methods that are faster and, because they are far more sensitive, require
only very small quantities of antigen or antibody. Also, these modern assay methods are not
limited to antigen-antibody reactions that produce a precipitate.

3: Neutralization Test:

It is an antigen-antibody reaction in which the harmful effects of bacterial exotoxin or virus are
blocked by specific antibodies. The antitoxin combines with the exotoxin to neutralize
it.Antitoxins produced in animals can be injected in humans to provide passive immunity against
the toxin. A more frequently used neutralization test is the viral heme agglutination inhibition
test. This test is used in the diagnosis of influenza, measles, mumps and a number of other
infections caused by viruses that can agglutinate RBC’s.

If a person serum contains antibodies against these viruses, antibodies will react with the viruses
and neutralize them e.g. if heme agglutination occurs in a mixture of measles virus and red blood
cell but does not occur when the patient serum is added in the mixture. Result indicate that serum
contain antibody that have bound to and neutralize the measles virus.
Complement-Fixation Reactionsnhnkrnaa

During most antigen- antibody reactions, complement binds to the antigen-antibody complex and
is used up, or fixed. This process of complement fixation can be used to detect very small
amounts of antibody. Antibodies that do not produce a visible reaction, such as precipitation or
agglutination, can be demonstrated by the fixing of complement during the antigen- antibody
reaction. Complement fixation was once used in the diagnosis of syphilis (Wassermann test) and
is still used to diagnose certain viral, fungal, and rickettsial diseases. The complement-fixation
test requires great care and good controls, one reason the trend is to replace it with newer,
simpler tests.
The test is performed in two stages: complement fixation and indicator
ELISA

The enzyme-linked immunosorbent assay (ELISA) is the most widely used of a group of tests

known as enzyme immunoassay (EIA ). There are two basic methods. The direct ELISA detects

antigens, and the illdirect ELISA detects antibodies. A microtiter plate with numerous shallow

wells is used in both procedures Variations of the test exist; for example, the reagents can be
bound to tiny latex particles rather than to the surfaces of the microtiter plates. ELISA

procedures are popular primarily because they require little interpretive skill to read; the results

tend to be clearly positive or clearly negative

VACCINATION

Vaccine is derived from Latin word VACCA means Cow. Vaccination is artificial introduction
of active immunity by administration of non-pathogenic form of disease causing agent.
Edward Jenner was founder of vaccination. It is very easy and cost affecting way of preventing
illness. It is also called prophylactic immunization.

VACCINE

Vaccine is a formulation containing pathogenic agents modified to make them non-pathogenic

and administration of which induce immune response in the recipient sufficient to prevent
susceptible disease.

OR

Vaccine is the suspension of attenuated or killed micro-organisms or their antigenic portion

presenting to a potential host to induce and immunologically mediated resistance to disease.

PRINCIPLE OF VACCINATION

To induce in an individual a prime state that on contact with relevant infection a more rapid and
effective immune response could be generated leaving toprevention of disease.

Vaccination depends on the ability of both B & T lymphocyte to respond to specific antigen and
develop into memory cell and this represent a form of actively enhanced adopted immunity. The
primary aim is to eliminate the disease.

PROPERTIES OF AN IDEAL VACCINE

1: Non-pathogenic:

A vaccine should not cause disease.

2: Immunogenicity:

A vaccine should be strongly immunogenic to induce high concentration of antibodies.

3: Immunogenic specificity:

It should produce antibodies that react specifically with disease producing agent.

4: Effectiveness:

It should be effective after a single dose.

5: Long lived immunity.

6: Low cost.

7: Compatibility to co-administer vaccine.

8: Stability:
It should be stable (genetically and thermal stability).

9: Response:

It should induce a wide range of appropriate responses (humoral, cellular and mucosal
immunity).

Types of Vaccine:

There are three types of vaccine.

1: First Generation Vaccine.

2: Second Generation vaccine.

3: Third Generation Vaccine.

First Generation Vaccine:

Those vaccines which are prepared from whole organisms whether live attenuated or killed
(bacterial or viral) are first generation vaccines.

A: Live attenuated vaccines:

Attenuation is a process of reducing virulence or toxic properties of micro-organism. Aim of

attenuation is to decrease toxicity and maintain antigenicity.

Methods to achieve attenuation:

1: Growing the organism at low temperature e.g. attenuation of influenza virus by growing at
low temperature.

2: By passing through unnatural host e.g. polio virus was attenuated by passage through monkey
kidney cell.

3: By treatment with mutagenic.

DISADVANTAGES:

1: Probability of reversal of attenuation.

2: They should not be given to immune-compromised persons.

3: Difficult and expensive to develop due to need to prove that micro-organism is non-
pathogenic.

ADVANTAGES:
 It gives strong and long lasting immunity due to amplification of immunogenic stimulant by
growth of micro-organism in the body.

B: Killed Vaccines:

Here pathogens are killed in such a way that the pathogenicity is reduced and terminated and
antigenicity is maintained.

METHODS:

1: By heating at 560C for 1 hour.

2: By treatment with chemical such as phenol 0.5% for bacterial vaccine.

3: β-propiolactone for viral vaccine.

ADVANTAGES:

1: These are relatively safe.

2: Relatively easy and cheap to produce.

3: Their safety testing is simpler.

DISADVANTAGES:

1: Immunity is weak and short lived as these organisms have no capacity to multiply so boaster
dose is essential for killed vaccine.

2: Allergic reactions may occur in some patient.

Second Generation Vaccines

Those vaccines which consist of defined naked or recombinant component derived from
organisams by biochemical purification or Genetic engineering.

1)Purified macromolecules as vaccines

In this case specific molecules are used as vaccines Forms of such vaccine include

1)Inactivated Exotoxins (Toxoid Vccine)

1)Capsular polysaccharide or polypeptide

Toxoid Vaccine
Some bacterial pathogens produce exotoxins such as Cornebacteriumdiptheriaeand Clostridium
tetani causing diphtheria and tetnus. These exotoxins can be inactivated with formaldehyde to
form toxoid. In the production of antisera conditions are so adjusted to achieve detoxification
without excessive modification of epitope structure.

Vaccination with toxoid inducesantitoxoid antibodies which are capable of binding to toxin and
neutralizing the toxic effects.

Advantage

Reduced pathogenesity as specific purified macromolecules derived from pathogens are used

ii) Recombinant antigen

Recombinant DNA technology has paved the way for development of new generation vaccine.

ANumber of genes encoding surface antigen from bacteria,virus and protozon pathogens have
been successfully cloned in bacteria, yeast, insect or mammalian expression system and
expressed antigens are used for vaccine development . Ist recombinant antigen vaccine produced
is Hepatitis B vaccine .Now a number of vaccines have been developed by this technology

Third Generation Vaccine

This include Nucleic acid vaccine. Introduction of Naked nucleic acid in vivo as a plasmid for
the purpose of generating an immune response is called a naked DNA vaccine. I t is the recent
technology in the field of vaccine development. It involves

 Isolation of DNA from infectious agent bacteria, virus, fungi and protozoa
 Induce eukaryotic promoters in the form of gene to DNA
 Introduction in Plasmid (Vector)
 Injection into host cell
 Intramuscular injection into skeletal muscle cardiac or diaphgram

Advantages

1) Easy to produce large quantity of pure DNA within short period of time

2) Safest no chances of reversal

3) Highly stable