Immunology

What is Immunology?

Immunology is the study of all aspects of the immune system, including its structure & function
(identifying & killing pathogens & tumor cells), disorders of the immune system (autoimmune
diseases, hypersensitivities, immune deficiency), blood banking, immunization, organ
transplantation & laboratory techniques based on antigen-antibody interaction.

Immunity

Immunity may be defined as the capacity of the body to resist or overcome infection.

The state of resistance may be manifested in either of two ways:

1. Not to develop infection despite exposure to a pathogen

2. To produce antibodies or immune-lymphoid cells against the pathogen.

Types of Immunity:

Immunity

Natural/ Innate/ Inherent/ Constitutional Acquired Immunity

Immunity

Active Immunity Passive Immunity

Species Race Individual

Natural Immunity:

This is the resistance to infection than can an individual processes inherently as part of the
genetic constitutional make up. It differs between species, races & individuals.

For Examples:

Species Differences: Men are susceptible to anthrax & tuberculosis but dogs are not. Typhoid
fever is a serious disease in human but mice are not affected.

Racial Differences: Negroes have a high resistance to yellow fever but white men are very
susceptible. The Caucasian race is more resistant to tuberculosis than
Negroes or American Indians.

Individual Differences: Some people are more resistant than others to cold & skin infections.

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Acquired Immunity:

This is the resistance to infection than an individual acquires during his life time.

Acquired Immunity is sub-divided into two types.

a) Active Immunity

b) Passive Immunity

a) Active Immunity: This is the immunity that an individual acquires through production of
antibodies and/or activated T-cells in response to an infectious agent or its antigen. This type of
immunity is relatively long-lasting. This can be naturally acquired or artificially stimulated.
Active immunity may further be sub-divided into two sub-types:

i. Naturally Acquired Active Immunity:

Following Clinical Infection:

With certain diseases, a single attack confers the life-long immunity. Examples include:
diphtheria, whooping cough, typhoid fever, scarlet fever, yellow fever & most viral diseases.

With some other disease, immunity is achieved for a short duration. Examples include:
Influenza, Pneumonia, Gonorrhea etc.

After Sub-clinical infection:

Because of the more frequent sub-clinical attack by the causative agents, children in slum areas
develop immunity to a variety of disease more quickly than children in affluent areas.

ii. Artificially Acquired Active Immunity:

This type of immunity is achieved by administration of antigenic preparations (containing killed

or attenuated microorganisms or their products)

Comparison between Active and Passive Immunity:

Active immunity Passive immunity

It is the immunity that develops when an
individual produces antibodies and/or activated
T-cells in response to an infectious agent or its
antigen.
It develops slowly and normally it takes
several months to be fully effective. (The
underlying reason is that training of the
antibody-producing cells takes considerable
time.)
It is long lasting and often gives protection for
many years.
It is the immunity that develops when an
individual receives antibodies of
immunolymphoid cells produced in an animal
or some other person.
It can be established relatively quickly. (The
time depends on the route of administration
may range from immediate for I/V injection to
perhaps 48 hours for S/C injection.)
It is very short lived. The protection is lost
within 2 to 3 weeks. (The underlying reason is
that the body recognizes the antibodies as

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 foreign and gradually eliminates them. Thus
when human antibodies are used, the rate of
elimination is much slower.)
Active immunization is used for the prevention Passive immunization is used both
of diseases prophylactically (to give immediate but
temporary protection to individuals exposed to
infection) and therapeutically (to treat an acute
infection).
Naturally stimulated by clinical or sub-clinical Naturally acquired from mother and artificially
infection and artificially produced by vaccines. produced by sera.

Immune system: Active immunity can be further divided into two types

 Innate (non-specific) Immune system

 Adaptive (specific) immune system

Innate Adaptive

1. Antigen independent 1. Antigen dependent

2. Immediate (hours) 2. Slower (days)

3. Can attack healthy cell 3. Wont attack Healthy cell

 Neutrophil  B cell

 Natural killer cell  T cell

 Macrophages

Adaptive immune system:

Humoral immunity is mediated by molecules in the blood and mucosal secretions, called
antibodies, that are produced by cells called B lymphocytes (also called B cells). Antibodies
recognize microbial antigens, neutralize the infectivity of the microbes, and target microbes for
elimination by various effector mechanisms.

Cell-mediated immunity, also called cellular immunity, is mediated by T lymphocytes (also

called T cells). Intracellular microbes, such as viruses and some bacteria, survive and proliferate
inside phagocytes and other host cells, where they are inaccessible to circulating antibodies.
Defense against such infections is a function of cell-mediated immunity, which promotes the
destruction of microbes residing in phagocytes or the killing of infected cells to eliminate
reservoirs of infection.

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Humoral Cell mediated

1. B lymphocytes mediated 1. T cell mediated

 Helper T cell- activated macrophage

 Cytotoxic cell- CD8 cell mediated cell killing

2. Action produced through antibody 2. Action produced through Cytokine production

production

3. Extracellular microbe. E.g. bacteria 3. Intracellular microbe. E.g. virus

Markers of self:

Major histocompatibility complex (MHC), group of genes that code for proteins found on the
surfaces of cells that help the immune system recognize foreign substances. MHC proteins are
found in all higher vertebrates. In human beings the complex is also called the human leukocyte
antigen (HLA) system. There are two classes- MHC class I proteins and MHC class II proteins.

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MHC class I protein MHC class II protein

1. Present on all nucleated cell 1. Only on specialized cell. E.g. Macrophage.

Lymphocytes

2. Present endogenous antigen 2. Present exogenous antigen

3. Recognized by CD8 cell 3. Recognized by CD4 cells

Structure of the Immune System:

Two Components:

a. Organs of the immune system

b. Cells of the immune system

a. Organs of the immune system:

i. Bone marrow:

 It is the initial generation site of all the cells of the immune system.

 It produces β-cells (B-lymphocytes), natural killer cells, granulocytes & immature

thymocytes (pro-thymocytes)

ii. Thymus:

 It produces mature T cells from immature thymocytes

iii. Spleen:

 Act as an immunologic filter of the blood.

 It is made up of β cells, T cells, macrophages, dendritic cells, natural killer cells & red
blood cells.

iv. Lymph Nodes:

 An immunologic filter for the lymph.

 Are composed mostly of T cells, B cells, dendritic cells & macrophages.

b. Cells of the immune system:

i. T cells:

T Cell: Two types- CD4 & CD8 cells.

 The differentiation of CD4 and CD8 occurs during their development in thymus

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CD4 helper T cell and accessory molecule:

 Recognize MHC class II protein

 Two types- T Helper Cell 1 & T Helper cell 2

Th 1 Th 2

 Bind with APC release IFN
and IL2
 IFN switch on cellular mode
by activating macrophages.
 Release IFN which send halt
signal to proliferation and
maturation of B cell and thereby
shut down humoral pathway.
 Binds with B cell and release
IL4 and IL5, IL13 and cause
proliferation and maturation of
B cell.
 Release antibody and activates
humoral pathway.
 Release IL10 which send halt
signal that down regulate
cellular pathway.

CD8 cytotoxic T cell:

 Recognize MHC class I protein.

 Can kill cell like other cytotoxic cell by death receptor (Ras TNFR) or by
granzymes and perforins

 Release cytokines usually TH1 types.

Perforins- punch holes in target cell followed by Granzymes that cause the cell to undergo
apoptosis

ii. Natural Killer cells (NK cells):

 Similar to the killer T cells (CD8 + T cells) directly kill certain tumors & viral infected
cells.

iii. β cells:

 Produce antibodies in response to foreign protein of bacteria, viruses & tumor cells.

iv. Granulocytes/ Polymorphonuclear (PMN) leukocytes:

 Engulf & enzymatically degrade bacteria & parasites.

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 v. Macrophages:

 Scavengers or antigen-presenting cells (APC) pick up & ingest foreign materials &
present their antigens to T cells & B cells.

vi. Dendritic cells:

 Function as antigen- presenting cells (APC).

Antigen presenting cells:

 Dendritic cells  B lymphocytes

 Natural killer cell  Monocytes

 Macrophage

A classification of microorganism on the basis of Host-Microbe Relationship:

Microorganism

Pathogens (Microorganism capable of Non-pathogens (Microorganisms

producing disease) unable to produce any disease)

Saprophytes (Microorganisms Commensals (Microorganisms living

growing on dead/organic matter) in association of a host without
causing any harm to the host)

INFECTION

What is Infection?

An infection is said to have occurred when a pathogenic microorganism invades a body part or
tissue of a host and starts multiplying and in turn causes tissue injury & overt disease through a
variety of cellular or toxic mechanism.

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Infectious Disease:

Diseases caused by pathogenic invasion, subsequent growth & multiplication of microorganisms

in the body are known as infectious diseases.

Modes of Transmission of Infection:

Infection may take place by:

1. Physical Contact with a Diseased Person or Animal:

 Means direct contact without intervention of any other objects.

 For example: Transmission of Rabies through the bite of an infected dog, transmission of
sexual /venereal disease like syphilis & gonorrhea through sexual contact.

2. Contact with Contaminated Articles:

 Clothes, bedding, handkerchiefs, towels, toys and other items recently contaminated by a
diseased person are sources of infection, either through contact or via the microbe-laden
particles discharged from them, into the air, by movement.

 Examples: Transmission of diphtheria from a child to another by means of a doll.

3. Hand Infection:

 The hands are a major potential means by which micro-organisms from saliva, nasal
mucus, skin infections, and even feces are transferred to other individuals and to food.

4. Droplet Infection:

 Coughing, sneezing and even quiet talking cause expulsion from the respiratory tract of
fine droplets containing micro-organisms which may be inhaled by individuals nearby.

 By this mechanism respiratory diseases are often transmitted. For example: whooping
cough, tuberculosis, diphtheria, cerebral meningitis, common cold, measles etc.

5. Dust-borne Infection:

 Some disease-producing bacteria, e.g. Mycobacterium tuberculosis, remain alive for long
periods in the dried condition, and contaminated dust stirred up by cleaning processes
may infect wounds or be inhaled.

6. Arthropod Vectors:

 Some disease-producing organisms are transmitted by insects (e.g. flies and mosquitoes)
and other arthropods (e.g. fleas and ticks).

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  By Arthropods feeding on fecal matter & food: In certain cases the vectors simply act
as accidental carriers of the organisms from filth to food or to the human host. In this
way, plague and typhoid fever are transferred by fleas and house-flies respectively.

 By Blood sucking Arthropods: In other cases, the micro-organisms are transmitted by

blood-sucking arthropods and spend part of their life cycle in the vector. Examples are
the transmission of malaria by the Anopheles mosquito, yellow fever by the Aedes
mosquito, sleeping sickness by the tsetse fly, and murine typhus by the rat flea.

Routes of Infection/Portals of entry:

The routes through which microorganism gain entrance to the body are as follows:

1. Ingestion: Ingestion of contaminated foods and drinks may cause infection. Examples
include typhoid fever, food poisoning, dysentery, cholera etc.

2. Inhalation: Inhalation of contaminated dusts, droplets, etc. may cause infection.

Examples include tuberculosis, cold, whooping etc.

3. Inoculation: (Introduction of microorganisms, infective material, serum or other

substances into tissues of living organisms or culture media) Intact skin offers resistance
against bacterial infections.

 Microorganisms usually enter through some break in continuity of skin e.g.

minute abrasions, lacerations, punctures etc.

 However, some organisms may penetrate intact skin. Examples include:

Treponema pallidum and Leptospira icterohaemorrhagia.

 Some other organisms enter across the skin through insect bites.

Factors Influencing Infection:

Pathogenicity: The ability of a microorganism to cause infection is called pathogenicity. The

factor influencing pathogenicity includes:

1. Virulence:

Virulence is the degree of pathogenicity of a pathogen. It has got two aspects:

i. Invasiveness: Invasiveness/aggressiveness is the ability of a microorganism to

invade- that is to enter the body and spread in the tissues.

ii. Toxigenicity: Toxigenicity is the capacity of a microorganism to produce toxins.

 Organisms possess these two qualities to varying degrees. For example: Clostridium
tetani can enter the body only through an injury. This means that its invasiveness is low.
But, once established, it produces a very powerful toxin, i.e. it has very high toxigenicity.

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  On the other hand, Sterptococcus pneumoniae enter the body easily and multiply readily,
but it has very limited ability to produce toxin.

Enzymes Associated with Virulence:

Some bacteria may liberate one or more enzymes or similar substances that facilitate invasion by
the organism or increase the damage caused by the toxin.

1. Hyaluronidase: This enzyme, which is also known as the spreading factor, reversibly
catalyses the breakdown of hyaluronic acid, a major component of the cement between
tissue cells. As a result, the viscous cement becomes fluid and bacteria and their toxins
can more easily spread throughout the body.

2. Collagenase: Collagenase disintegrates collagen, a constituent of muscle, cartilage and

bone.

3. Lecithinase: An important constituent of cell membranes is lecithin. Lecithinase activity

results in the haemolysis of erythrocytes and the necrosis of other cells.

4. Coagulase: Certain highly pathogenic strains of Staphylococcus aureus produce this

enzyme which clots plasma and surrounds the bacteria with a barrier of fibrin that
protects them against phagocytosis.

5. Haemolysins: Many bacteria, e.g. streptococci and clostridia, produce enzymes or other
substances that destroy red blood cells.

6. Leucocidins: The body has a number of lines of defence against infection. Leucocytes
(white blood cells) have important roles in the body's defences against bacteria.
Leucocidins inactivate or destroy them and so aid bacterial invasion.

Changes in Virulence:

Since natural differences in virulence are found in different strains of bacteria it is essential to
use strains of the correct virulence for the manufacture of immunological products.

Virulence may be artificially altered thus:

1. Increased (exalted): By successive fairly rapid transfers from one susceptible host to another.

2. Decreased (attenuated):

(a) By growing the organism under artificial conditions in laboratory culture media. Usually an
organism is most virulent when first isolated from its host, and unless it is grown on a medium
that closely simulates its natural environment (e.g. a medium well enriched with sources of
animal protein) it will often lose virulence (e.g. BCG vaccine).

(b) By growing the organism at an unfavourable temperature. (e.g. Pasteur's work on anthrax
vaccine.)

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(c) By growing the organism in an unnatural host - (e.g. Yellow fever vaccine). Virulence for
man may be lost after repeated passages through laboratory animals.

(d) By drying (e.g. Rabies vaccine).

2. The Number of Organisms:

The number of organisms required to produce disease varies with-

 The species and resistance of the host

 The species or strain and virulence of the infecting organisms.

However, in general, the larger the number of organisms, the greater the chance of establishment
of infection.

[The number of organisms required to cause death or a characteristic sign of infection is

sometimes used as an indication of virulence in immunological preparations.]

3. The Route of Entry:

In many diseases, the route of entry is important for development of infection. That is, infection
will not occur unless the organisms enter the body by particular route such as alimentary tract for
typhoid, genitourinary treat for gonorrhea and respiratory tract for diphtheria.

4. Resistance of the Host:

The body has a number of lines of defense against infection including:

 Prevention of entry

 Phagocytosis

 Production of antibodies and activated T lymphocytes

a. Prevention of Entry:

Our body has a number of devices by which it prevents the entry of microorganisms. These
include:

 The design of the body structures. For example, the nasal passage can effectively trap
microorganisms.

 The impermeability of the intact and healthy skin and mucosae to microorganisms.

 Competition from the natural microbial flora of the host.

 The antimicrobial activities of the secretions of the body surfaces. For instance:

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  The mucus covering of mucus membranes collects and retains microorganisms,
which are then expelled by ciliary action aided by coughing and sneezing in the
respiratory tract or by peristaltic activity in the gut.

 The pH of the gastric juice

 The fatty acids in the skin

 The enzyme lysozome in tears and sweat

All of these can inhibit or destroy microorganisms.

b. Phagocytosis:

Phagocytosis is nonspecific defense mechanism that is directed against all microorganisms. It

involves engulfment of microorganism by white blood cells. The engulfing cells are known as
phagocytes. There are two types of phagocytes: fixed and wandering.

 Fixed phagocytes include macrophages (lining blood spaces in different parts of body
specially spleen and liver).

 The wandering phagocytes include neutrophils. (These agents pass from the blood
across the capillary wall to the site of infection in response to inflammatory changes
caused by microorganisms.)

c. Production of Antibodies and Activated T-cells:

This is a specific defense mechanism. That is, antibodies or activated T cells produced in
response to a certain microorganism will specifically act against that microorganism and no
other. There are a number or mechanisms by which antibodies can destroy or inhibit
microorganisms.

Antigen:

Antigens are proteins or large polysaccharides that on entering to the body, initiate development
of acquired immunity (i.e. production of specific antibodies or activated T-cells, which react with
antigenic substances triggering their production).

Usually for a substance to be antigenic it must have a higher molecular weight (8000 or more).
Antigenic substance also contains regularly recurring molecular groups called epitopes on their
surface.

Anitbody:

Antibodies are a group of proteins, which are produced in the body in response to introduction of
antigenic substance and which interact in a specific, antagonistic manner only with the antigens
inducing their production. Antibodies are mostly Ƴ-globulins and are also known as
immunoglubulins, abbreviated as Ig. On the basis of structure and biologic activity, they are
divided into five general classes: IgM, IgG, IgA, IgD and IgE.

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IgG→ Constitute 75% of antibodies of a normal individual.

IgE (Reaginic antibodies)→ Constitute only a small percentage of antibodies, but they mediate
allergic reactions.

IgM→ Have 10 binding sites and is highly effective in protecting body against invaders.
Constitute a large share of antibodies formed during primary response.

Direct action of Antibodies on the Invading Agent

Antibodies can directly inactivate invading agents in a number of ways:

1. Agglutination: In this case, the antibodies bind together multiple antigen containing large
particles (such as bacteria or red blood cells) into a clump.

Agglutination delays disintegration, and consequent release of endotoxins from cells. It also
facilitates phagocytosis of the cells by decreasing their motility.

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The antigen involved in this reaction is known as agglutinogen and the antibody agglutinin.

The antibody involved in this reaction is usually of IgM type.

2. Precipitation: In this case, the antibody forms a large insoluble complex with a small,
soluble antigen (e.g. tetanus toxin) and thereby precipitates it. Liberated endotoxins are
inactivated by this reaction.

The antigen involved in this reaction is known as precipitinogen and the antibody as precipitin.

3. Neutralization: In this case, the antibody covers the toxic sites of antigenic agent. Bacterial
exotoxins are rendered harmless by this mechanism.

The antibody participating in this reaction is known as antitoxin.

4. Lysis: Some potent antibodies can directly attack the cell membrane causing rupture of the
cell.

Antigen binding
site

Bacterial Toxins:

There are two types of bacterial toxins: Exotoxins and Endotoxins.

Difference between Exotoxins and Endotoxins:

Exotoxins Endotoxins

Exotoxins are the bacterial toxins that diffuse Endotoxins are the bacterial toxins that are
freely through the bacterial cell wall into the retained within the bacteria and are not
surrounding media (and can be separated from released until the cells die and start to
the cells by bacteria-proof filter) disintegrate.

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They are metabolic products of actively They are structural elements of bacteria.
growing bacteria

They are produced mainly by gram +ve They are produced more or less by all bacterial
bacteria cells, but found in large amount only in gram

–ve bacteria.

They are water-soluble and can pass into the They are not excreted into the surrounding
surrounding medium. This provides a means of medium and are liberated only when the cells
separating the toxin from the cells, since the die or disintegrate.
latter can be removed on a bacteria-proof filter
through which the toxin-containing medium
will pass.

Chemically they are high molecular weight Chemically, they are complexes of
proteins and some are enzymes. Thus they are phospholipid, polysaccharide and protein, most
relatively thermolabile and lost their activity at of which are thermostable.
about 600C

Usually they are extremely toxic to the body They are much less toxic.

Examples: Clostridium tetani (Tetanus), Examples: Vibrio cholerae (Cholera),

Clostridium botulinum (Food poisoning) Salmonella typhi (Thyphoid)

Comparison between Vaccines and Sera:

Vaccines Sera

Antigenic preparations. Antibody containing preparations.

Stimulate active immunity. Give passive immunity.

Patient produces antibodies. Patient receives antibodies.

Immunity develops slowly. Immunity is produced quickly.

Give lasting effect. Give temporary effect.

Used for long-term prophylaxis. Used for short-term prophylaxis and

therapeutically.

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