MODULE 3

IMMUNE SYSTEM IN HEALTH AND

DISEASE
IMMUNE SYSTEM IN HEALTH AND DISEASE:

Complement system and its pathways, activation and function.

Hypersenitivity: Gell and Coombs classification of Hypersensitivity reactions and Diagnosis
and treatment.
Autoimmune disorders, types, animal model and treatment.
Immune response to infections: immunity to viruses, bacteria, fungi and parasites,
Immunodeficiency disorders: Primary and secondary (AIDS).
Injury and inflammation,
Vaccines and their types, classification and
immunization schedule
Production of recombinant vaccine for hepatitis B surface antigen

Dr. Amulya Giridasappa, The Oxford College of Engineering

Inflammation – It is a complex sequence of events induced during a tissue damage caused by a wound or invasion
by a pathogenic micro-organism. Inflammation has four characteristic signs – redness, swelling, heat & pain.
These signs of inflammation reflect three major events which occur during the inflammatory
response.
➢ Vasodilation
➢ Increased capillary permeability
➢ Influx of phagocytic cells & serum proteins having antibacterial properties into tissue
1. Vasodilation is an increase in the capillary diameter. It occurs as the blood vessels which carry blood away
from the affected area constrict resulting in bulging of capillary network.
• The swollen capillaries are responsible for tissue redness & an increase in tissue temperature. As a result, an
increase in capillary permeability occurs, which helps in influx of fluid & cells from engorged capillaries into the
tissue.
• The fluid that accumulates in tissue is rich in protein content, which is higher than the protein content of the fluid
which is normally released from the vasculature. Accumulation of fluid results in the tissue swelling.

Dr. Amulya Giridasappa, The Oxford College of Engineering

The increased capillary permeability also helps in the migration of various white blood cells especially phagocytic
cells from blood capillaries into tissue. These proteins & phagocytic cells help to eliminate the foreign cells &
materials from the tissue, which can enter the body through the wound.

Complement
“Complement is a group of serum proteins which circulate in the serum in an inactive proenzyme form. These
inactive proteins can be activated by various specific & nonspecific immunologic mechanisms. The activation of
complement molecule helps in the formation of Membrane Attack Complex (MAC) & other smaller peptides”.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• These help in elimination of microorganisms from our body either by damaging the cell membrane of
pathogenic microorganisms or by facilitating their clearance. Complement acts as the effector of the humoral
branch of the immune system.
• The proteins and glycoproteins that compose the complement system are synthesized mainly by liver
hepatocytes, although significant amounts are also produced by blood monocytes, tissue macrophages, and
epithelial cells of the gastrointestinal and genitourinary tracts. These components constitute 5% (by weight) of
the serum globulin fraction.
• Complement components are designated by numerals (C1–C9), by letter symbols (e.g., factor D), or by trivial
names (e.g., homologous restriction factor). Peptide fragments formed by activation of a component are
denoted by small letters. In most cases, the smaller fragment resulting from cleavage of a component is
designated ―a‖ and the larger fragment designated ―b‖ (e.g., C3a, C3b; note that C2 is an exception: C2a is
the larger cleavage fragment). The larger fragments bind to the target near the site of activation, and the
smaller fragments diffuse from the site and can initiate localized inflammatory responses by binding to specific
receptors. The complement fragments interact with one another to form functional complexes. Those
complexes that have enzymatic activity are designated by a bar over the number or symbol (e.g., C4b2a,
C3bBb). Dr. Amulya Giridasappa, The Oxford College of Engineering
Complement Activation - Inactive proenzyme form of complement can be activated by two different pathways.
They are
• Classical pathway
• Alternative pathway
Both pathways follow different mechanism to produce the C5b. The final steps that lead to a membrane attack
complex are same in both the pathways.

Classical pathway
Complement activation by the classical pathway commonly begins with the formation of soluble antigen-antibody
complexes (immune complexes) or with the binding of antibody to antigen on a suitable target, such as a bacterial
cell. IgM and certain subclasses of IgG (human IgG1, IgG2, and IgG3) can activate the classical complement
pathway. The initial stage of activation involves C1, C2, C3, and C4, which are present in plasma in functionally
inactive forms.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Classical Pathway:
Initiation: Triggered by the binding of antibodies (IgM or IgG) to antigens on the surface of pathogens.
C1 Complex Activation: The C1 complex, composed of C1q, C1r, and C1s, binds to the antibody-antigen
complex, leading to its activation.
Cleavage Cascade: Activated C1 cleaves C4 and C2, generating C4a, C4b, C2a, and C2b.
C3 Convertase Formation: C4b and C2a combine to form C3 convertase, which cleaves C3 into C3a and C3b.
Amplification: C3b is an opsonin that coats the pathogen surface, enhancing phagocytosis. C3b also participates
in the formation of the C5 convertase.
***************************
The classical pathway of the complement system is one of the three main activation pathways (alongside the
alternative pathway and the lectin pathway). The classical pathway is primarily initiated by the binding of
antibodies (IgM or IgG) to antigens on the surface of pathogens. Here is an overview of the classical pathway:

1. Initiation: The classical pathway is initiated when antibodies (IgM or IgG) bind to specific antigens on the
surface of pathogens. This antibody-antigen interaction is a key event that marks the pathogen for destruction.

Dr. Amulya Giridasappa, The Oxford College of Engineering

2. C1 Complex Activation:
• The C1 complex is composed of three subunits: C1q, C1r, and C1s.
• Binding of the C1q component to the Fc region of antibodies in the antibody-antigen complex activates C1r
and C1s. This activation occurs through conformational changes in the C1q structure.
3. Cleavage Cascade:
• Activated C1s protease cleaves complement proteins C4 and C2, generating smaller fragments.
• C4 is cleaved into C4a and C4b.
• C2 is cleaved into C2a and C2b.
4. C3 Convertase Formation:
• The C4b and C2a fragments combine to form the C3 convertase (C4b2a). This enzyme is capable of cleaving
the central complement component, C3.
5. C3 Cleavage:
• The C3 convertase (C4b2a) cleaves C3 into two fragments: C3a and C3b.
• C3a is an anaphylatoxin that promotes inflammation.
• C3b plays a crucial role in opsonization.
Dr. Amulya Giridasappa, The Oxford College of Engineering
6. Amplification:
• C3b, the product of C3 cleavage, participates in the formation of the C5 convertase. It binds to C4b2a,
creating the C4b2a3b complex, which functions as the C5 convertase.
7. C5 Convertase Formation:
• The C5 convertase (C4b2a3b) cleaves the C5 complement protein into C5a and C5b.
8. Terminal Steps:
• The C5b fragment initiates the assembly of the membrane attack complex (MAC).
• The MAC consists of C5b, C6, C7, C8, and multiple C9 molecules. It forms a pore in the membrane of the
target cell, leading to cell lysis.
The classical pathway is particularly effective against pathogens that have been tagged with antibodies, and it
provides a rapid and specific response to infections. It is a crucial component of the overall complement system,
contributing to the elimination of pathogens and the modulation of immune responses.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Process 1. C1 in serum is a macromolecular complex consisting of C1q and two molecules each of C1r and C1s,
held together in a complex (C1qr2s2) stabilized by Ca2+ ions.
2. Each C1 molecule must bind by its C1q globular heads to at least two Fc sites for a stable C1-antibody
interaction to occur. The formation of an antigen-antibody complex induces conformational changes in the Fc
portion of the IgM molecule that expose a binding site for the C1 component of the complement system. Whereas
two IgG molecules bound close enough to each other on the cell surface can initiate C1q binding.
3. Binding of C1q to Fc binding sites induces a conformational
change in C1r that converts C1r to an active serine protease enzyme,
C1r, which then cleaves C1s to a similar active enzyme; C1s.
C1s has two substrates,C4 and C2. The C4 component is a
glycoprotein containing three subunits α, β, and γ. C4 is activated
when C1s hydrolyzes a small fragment (C4a) from the amino
terminus of the α chain, exposing a binding site on the larger
fragment (C4b).

Dr. Amulya Giridasappa, The Oxford College of Engineering

4. The C4b fragment attaches to the target surface in the vicinity of C1, and the C2 proenzyme then attaches to
the exposed binding site on C4b, where the C2 is then cleaved by the neighboring C1s; the smaller fragment
(C2b) diffuses away. The resulting C4b2a complex is called C3 convertase. C3 convertase converts the
inactive C3 component into active form.
5. The native C3 component consists of two polypeptide chains, α and β. Hydrolysis of a short fragment (C3a)
from the amino terminus of α chain by the C3 convertase generates C3b.
6. A single C3 convertase molecule can generate over 200 molecules of C3b, resulting in tremendous
amplification at this step of the sequence. Some of the C3b binds to C4b2a to form a trimolecular complex
C4b2a3b, called C5 convertase.
7. The C3b component of this complex binds C5 and alters its conformation, so that the C5 convertase‘s C4b2a
component can cleave C5 into C5a smaller fragment, which diffuses away, and C5b larger fragment.
8. The terminal sequence of complement activation involves C5b, C6, C7, C8, and C9, which interact
sequentially to form a macromolecular structure called the membrane-attack complex (MAC).
9. The C5b component binds to C6. Binding of C6 to C5b produces C5b6 bimolecular complex. C5b6 binds to
C7 to form C5b67. C5b67 undergoes a hydrophilic-amphiphilic structural transition that exposes hydrophobic
regions C5b67 complex, thus can insert into the phospholipid
Dr.bilayer.
Amulya Giridasappa, The Oxford College of Engineering
10. Binding of C8 to membrane-bound C5b67 induces a conformational change in C8, so that it too
undergoes same structural transition, exposing a hydrophobic region, which interacts with the plasma
membrane. The C5b678 complex creates a small pore, 10 Å in diameter.
11. The final step in formation of the MAC is the binding and polymerization of C9, a perforin-like molecule, to
the C5b678 complex.
12. As many as 10–17 molecules of C9 can be bound and polymerized by a single C5b678 complex.
13. During polymerization C9 molecules undergo a hydrophilic-amphiphilic transition, so that they too can
insert into the membrane. The completed MAC, which has a tubular form and functional pore size of 70–
100 Å, consists of a C5b678 complex surrounded by a poly-C9 complex.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Alternative Pathway:
•Initiation: Triggered by spontaneous hydrolysis of C3 in the absence of antibodies.
•C3 Cleavage: C3 undergoes spontaneous hydrolysis, producing C3a and C3b.
•Factor B and Properdin: C3b binds to factor B, and the complex is stabilized by properdin.
•C3 Convertase Formation: The stabilized C3bBb complex acts as a C3 convertase, leading to further cleavage
of C3 into C3a and C3b.
•Amplification: Similar to the classical pathway, C3b participates in the formation of the C5 convertase.
***************************
The alternative pathway is one of the three main pathways of the complement system, along with the classical
pathway and the lectin pathway. The alternative pathway is unique because it can be activated spontaneously
without the need for antibodies. It serves as a crucial component of the innate immune response. Here is an
overview of the alternative pathway:
1. Initiation (Spontaneous Hydrolysis):
• The alternative pathway is initiated by the spontaneous hydrolysis of C3 (complement protein 3) in the
bloodstream.
• A small fraction of C3 molecules undergo spontaneous hydrolysis, resulting in the formation of C3(H2O).
Dr. Amulya Giridasappa, The Oxford College of Engineering
2. Factor B Binding:
• Once C3 is hydrolyzed to C3(H2O), it can bind to complement factor B.
3. Stabilization by Properdin:
• The C3bB complex (formed by the binding of C3(H2O) to factor B) is stabilized by the protein properdin (P).
• Properdin helps prevent the rapid decay of the C3 convertase.
4. C3 Convertase Formation:
• The stabilized C3bB complex can recruit and bind another molecule of factor B, leading to the formation of the
alternative pathway C3 convertase, known as C3bBb.
5. C3 Cleavage:
• The alternative pathway C3 convertase (C3bBb) cleaves additional C3 molecules into C3a and C3b.
6. Amplification:
• Similar to the classical pathway, C3b produced by the alternative pathway can participate in the formation of
the C5 convertase.
7. C5 Convertase Formation:
• C3b, in conjunction with other complement proteins, forms the alternative pathway C5 convertase, known as
C3bBbC3b. Dr. Amulya Giridasappa, The Oxford College of Engineering
8. Terminal Steps:
1. The alternative pathway C5 convertase (C3bBbC3b) cleaves the C5 complement protein into C5a and C5b.
9. Membrane Attack Complex (MAC):
• The C5b fragment initiates the assembly of the membrane attack complex (MAC).
• The MAC consists of C5b, C6, C7, C8, and multiple C9 molecules. It forms a pore in the membrane of the
target cell, leading to cell lysis.
The alternative pathway provides a rapid and broad-spectrum response to infections, as it can be activated
spontaneously by the presence of pathogens. It plays a critical role in the elimination of microorganisms and contributes
to the overall defense mechanisms of the immune system.
Alterntative pathway (Indetail) -
The alternative pathway generates bound C5b, the same product that the classical pathway generates, but it does so
without the need for antigen-antibody complexes for initiation. Because no antibody is required, the alternative
pathway is a component of the innate immune system. This major pathway of complement activation involves four
serum proteins: C3, factor B, factor D, and properdin. The alternative pathway is initiated in most cases by cell-
surface constituents that are foreign to the host. For example, both gram-negative and gram-positive bacterial cell-
wall constituents can activate the alternative pathway.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Process: 1. In the alternative pathway, serum C3, which contains an unstable thioester bond, is subject to slow
spontaneous hydrolysis to yield C3a and C3b. The C3b component can bind to foreign surface antigens (such as
those on bacterial cells or viral particles) or even to the host‘s own cells. The membranes of most mammalian cells
have high levels of sialic acid, which contributes to the rapid inactivation of bound C3b molecules on host cells;
consequently this binding rarely leads to further reactions on the host cell membrane.
2. Because many foreign antigenic surfaces (e.g bacterial cell walls, yeast cell walls, and certain viral envelopes)
have only low levels of sialic acid, C3b bound to these surfaces remains active for a longer time. The C3b present on
the surface of the foreign cells can bind another serum protein called factor B to form a complex stabilized by Mg2+.
3. Binding to C3b exposes a site on factor B that serves as the substrate for an enzymatically active serum protein
called factor D. Factor D cleaves the C3b-bound factor B, releasing a small fragment (Ba) that diffuses away and
generating C3bBb.
4. The C3bBb complex has C3 convertase activity and thus is analogous to the C4b2a complex in the classical
pathway. The C3 convertase activity of C3bBb has a half-life of only 5 minutes unless the serum protein properdin
binds to it, stabilizing it and extending the half-life of this convertase activity to 30 minutes. The C3 convertase can
activate unhydrolyzed C3 to generate more C3b autocatalytically.
5. As a result, the initial steps are repeated and amplified, so that more than 2 X 10^6 molecules of C3b can be
deposited on an antigenic surface in less than 5 minutes. Dr. Amulya Giridasappa, The Oxford College of Engineering
6. The C3 convertase activity of C3bBb generates the
C3bBb3b complex, which exhibits C5 convertase activity,
analogous to the C4b2a3b complex in the classical pathway.
The nonenzymatic C3b component binds C5, and the Bb
component subsequently hydrolyzes the bound C5 to
generate C5a and C5b; the latter binds to the antigenic
surface.
7. The terminal sequence of complement activation involves
C5b, C6, C7, C8, and C9, which interact sequentially to form
a macromolecular structure called the membrane-attack
complex (MAC).
8. The C5b component binds to C6. Binding of C6 to C5b
produces C5b6 bimolecular complex. C5b6 binds to C7 to
form C5b67. 9. C5b67 undergoes a hydrophilic-amphiphilic
structural transition that exposes hydrophobic regions C5b67
complex, thus can insert into the phospholipid bilayer.
Dr. Amulya Giridasappa, The Oxford College of Engineering
10. Binding of C8 to membrane-bound C5b67 induces a conformational change in C8, so that it too undergoes same
structural transition, exposing a hydrophobic region, which interacts with the plasma membrane. The C5b678
complex creates a small pore, 10 Å in diameter.
11. The final step in formation of the MAC is the binding and polymerization of C9, a perforin-like molecule, to the
C5b678 complex. As many as 10–17 molecules of C9 can be bound and polymerized by a single C5b678 complex.
During polymerization C9 molecules undergo a hydrophilic-amphiphilic transition, so that they too can insert into the
membrane. The completed MAC, which has a tubular form and functional pore size of 70–100 Å, consists of a
C5b678 complex surrounded by a poly-C9 complex.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Functions of complement
After initial activation, the various complement components interact, in a highly regulated cascade, to carry out a
number of basic functions, they are:
1. Lysis of cells, bacteria, and viruses
2. Opsonization, which promotes phagocytosis of particulate antigens
3. Binding to specific complement receptors on cells of the immune system, triggering specific cell functions,
inflammation, and secretion of immunoregulatory molecules
4. Immune clearance, which removes immune complexes from the circulation and deposits them in the spleen and
liver.
5. Chemotaxis- Complement fragments act as chemoattractants, guiding immune cells to the site of infection.

Dr. Amulya Giridasappa,

The Oxford College of
Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
Hypersensitivity Reactions
An immune response produces various effector molecules that act to remove antigen by various
mechanisms. Generally, these effector molecules induce a localized inflammatory response that eliminates antigen
without extensively damaging the host‘s tissue by producing subclinical manifestations. Under certain circumstances,
however, this inflammatory response can have deleterious effects and produce clinical manifestations associated
with significant tissue damage or even death. This inappropriate immune response is termed hypersensitivity or
allergy.
Hypersensitivity can be defined as an exaggerated immune response produced by the immune system
having clinical manifestations. Hypersensitive reactions may develop in the course of either humoral or cell-mediated
responses.
The hypersensitivity reactions initiated by humoral branch by antibody or antigen-antibody complexes are
called immediate hypersensitivity, because the symptoms are manifest within minutes or hours after a sensitized
recipient encounters antigen. The hypersensitivity reactions induced by cell mediated immunity is called Delayed-
type hypersensitivity (DTH) is so named in recognition of the delay of symptoms until days after exposure.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Gell and Coombs Classification
Philip. G. H. Gell and Robin. R. A. Coombs based on the mechanism of immune reaction proposed a classification
scheme in which hypersensitive reactions are divided into four types. They are:
• Type I Hypersensitivity Reactions
• Type II Hypersensitivity Reactions
• Type III Hypersensitivity Reactions
• Type IV Hypersensitivity Reactions
Among these Type I, Type II & Type III hypersensitivity reactions are induced by the humoral branch and are
mediated by antibody or antigen-antibody complexes. Type IV hypersensitivity reactions are caused by the cell-
mediated branch, and are termed delayed-type hypersensitivity, or DTH.

Type I Hypersensitivity: Involves an IgE-mediated immune response. Upon initial exposure to an allergen, B cells
produce IgE antibodies, which bind to mast cells and basophils. Subsequent exposure to the allergen leads to cross-
linking of IgE antibodies, triggering the release of histamine and other inflammatory mediators. It is rapid onset,
occurs typically within minutes of exposure.
Examples: Allergic rhinitis (hay fever), asthma, anaphylaxis, and some types of food allergies.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Type II Hypersensitivity (Cytotoxic Hypersensitivity): Antibodies, often IgG or IgM, target antigens on the surface
of cells, leading to cell destruction through complement activation or phagocytosis. This type is often associated with
autoimmune reactions. It can vary in onset.
Examples: Autoimmune hemolytic anemia, blood transfusion reactions, and certain drug-induced reactions.

Type III Hypersensitivity (Immune Complex-Mediated Hypersensitivity): Immune complexes (antigen-antibody

complexes) form in the bloodstream and deposit in tissues, leading to complement activation and an inflammatory
response. It is typically delayed, occurring hours to days after exposure to the antigen.
Examples: Systemic lupus erythematosus (SLE), rheumatoid arthritis, and certain forms of vasculitis.

Type IV Hypersensitivity (Delayed-Type Hypersensitivity): Involves activation of T cells, particularly CD4+ and
CD8+ T cells. T cell-mediated inflammation and tissue damage occur, usually 24 to 48 hours after exposure to the
antigen. I t has delayed onset, usually occurring 24 to 48 hours after exposure.
Examples: Contact dermatitis (e.g., poison ivy), tuberculin skin test reactions, and certain autoimmune diseases.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Dr. Amulya Giridasappa,
The Oxford College of
Engineering
Treatment of Type I hypersensitivity reactions

• Immunotherapy with repeated injections of increasing doses of allergens (hyposensitization) has been known for
some time to reduce the severity of type I reactions, or even eliminate them completely, in a significant number of
individuals suffering from allergic rhinitis. Such repeated introduction of allergen by subcutaneous injections
appears to cause a shift toward IgG production or to induce T-cell–mediated suppression (possibly by a shift
to the TH1 subset and IFN-γ production) that turns off the IgE response.
• Another form of immunotherapy is the use of humanized monoclonal anti-IgE. These antibodies bind to IgE, but
only if IgE is not already bound to FcεRI; the latter would lead to histamine release.
• Another approach for treating allergies stems from finding that soluble antigens tend to induce a state of allergy by
activating T cells in the absence of the necessary co-stimulatory signal.
• Several drugs block release of allergic mediators by interfering with various biochemical steps in mast-cell
activation and degranulation. Disodium cromoglycate (cromolyn sodium) prevents Ca2+ influx into mast cells.
Theophylline, which is commonly administered to asthmatics orally or through inhalers, blocks phosphodiesterase,
which catalyzes the breakdown of cAMP to 5′-AMP. The resulting prolonged increase in cAMP levels blocks
degranulation. Dr. Amulya Giridasappa, The Oxford College of Engineering
• A number of drugs stimulate the β-adrenergic system by stimulating β-adrenergic receptors. Epinephrine (also
known as adrenaline) & its analogs are commonly administered during anaphylactic shock. It acts by binding to β-
adrenergic receptors on bronchial smooth muscles and mast cells, elevating the cAMP levels within these cells.
• The increased levels of cAMP promote relaxation of the bronchial muscles and decreased mast-cell
degranulation.
• Cortisone and various other anti-inflammatory drugs also have been used to reduce type I reactions.

Treatment of Type II hypersensitivity reactions

There is no cure for these diseases, the treatment aims at symptom control only. Because of the pathogenesis of
these diseases are antibody in origin, a lot of treatment options are aimed at that.
Depending on the severity of the hypersensitivity reaction, different treatment approaches are applied. Treatment
options, either given alone or in combination, include the following:
• steroids: these drugs include prednisolone, dexamethasone, etc. In type II hypersensitivity diseases, sometimes
high dose steroids are used. Depending on the diseases, steroid could become a long-term medication. In such
cases, long term use will need medical supervision for monitoring of potential side effects.
Dr. Amulya Giridasappa, The Oxford College of Engineering
•intragam infusion: this is infusing the body with antibodies. There are many potentially severe side effects due to
this, hence it must be administered under specialist supervision.
•plasmaphoresis: this is removing the blood autoantibodies.
•other drugs: interferon, cyclophosphamide, cyclosporin.

Treatment of Type III hypersensitivity reactions

•Removal of the offending agent is the mainstay of treatment of type III hypersensitivity reaction.
•Antihistamines and nonsteroidal anti-inflammatory drugs can provide symptomatic relief.
•Corticosteroids are used in severe cases to suppress inflammation. They are also used as premedication to prevent
hypersensitivity from happening.
•SLE is treated based on the individual patient's disease condition. Hydroxychloroquine is essential for long-term
treatment in all SLE patients. Antimalarials, corticosteroids, nonbiologic DMARDS, nonsteroidal anti-inflammatory,
and biologic DMARDs are other medications used to treat SLE.
•Treatment of autoimmune disorders (e.g., SLE) includes one or a combination of hydroxychloroquine, NSAIDs,
azathioprine, cyclophosphamide, methotrexate, mycophenolate, and tacrolimus
Dr. Amulya Giridasappa, The Oxford College of Engineering
Treatment of Type IV hypersensitivity reactions
• Some common causes of type 4 hypersensitivity reactions include exposure to poison ivy, certain metals, and
drugs such as antibiotics or anticonvulsants.
• Treatment varies from case to case. With contact dermatitis, for example, a doctor may prescribe topical steroids.
However, with tuberculin-type hypersensitivity, the doctor will use a normal procedure for tuberculosis. Common
treatments for tuberculin-type hypersensitivity include:
•rifampin
•isoniazid
•pyrazinamide
•ethambutol

Dr. Amulya Giridasappa, The Oxford College of Engineering

Autoimmune disorders
Autoimmune disorders are conditions in which the immune system mistakenly targets and attacks the body's own
cells, tissues, and organs. Normally, the immune system is designed to identify and destroy foreign substances like
bacteria and viruses. However, in autoimmune disorders, the immune system becomes confused and starts
recognizing the body's own cells as foreign, leading to immune-mediated damage. Autoimmune disorders can affect
various organs and tissues, and they can range from mild to severe.
1.Rheumatoid Arthritis (RA):
• Affected Tissues: Joints
• Characteristics: Chronic inflammation of the joints, leading to pain, swelling, and joint damage.
2.Systemic Lupus Erythematosus (SLE):
• Affected Tissues: Multiple organs, including skin, joints, kidneys, heart, lungs, and blood cells.
• Characteristics: Affects various organs, leading to a range of symptoms such as skin rash, joint pain, kidney
dysfunction, and fatigue.
3.Type 1 Diabetes: Affected Tissues: Pancreatic beta cells
• Characteristics: Destruction of insulin-producing cells in the pancreas, resulting in insulin deficiency and high
blood sugar levels. Dr. Amulya Giridasappa, The Oxford College of Engineering
4. Multiple Sclerosis (MS):
• Affected Tissues: Central nervous system (brain and spinal cord)
• Characteristics: Demyelination of nerve fibers, leading to problems with coordination, balance, and cognitive
function.
5. Hashimoto's Thyroiditis:
• Affected Tissues: Thyroid gland
• Characteristics: Autoimmune inflammation of the thyroid gland, resulting in hypothyroidism (underactive
thyroid).
6. Graves' Disease:
• Affected Tissues: Thyroid gland
• Characteristics: Autoimmune stimulation of the thyroid gland, leading to hyperthyroidism (overactive thyroid).
7. Inflammatory Bowel Disease (IBD):
• Affected Tissues: Gastrointestinal tract (Crohn's disease and ulcerative colitis)
• Characteristics: Chronic inflammation of the digestive tract, causing symptoms such as abdominal pain,
diarrhea, and weight loss.
Dr. Amulya Giridasappa, The Oxford College of Engineering
8. Psoriasis:
• Affected Tissues: Skin
• Characteristics: Chronic autoimmune skin condition, resulting in the formation of thick, red, and scaly
patches.
9. Celiac Disease:
• Affected Tissues: Small intestine
• Characteristics: Immune reaction triggered by gluten consumption, leading to damage to the lining of the
small intestine.
10. Rheumatic Fever:
• Affected Tissues: Heart, joints, skin, and central nervous system
• Characteristics: Occurs as a complication of untreated streptococcal infections, leading to inflammatory damage
to various tissues.
11. Sjögren's Syndrome:
• Affected Tissues: Salivary and lacrimal glands
• Characteristics: Chronic inflammation that primarily affects the moisture-producing glands, leading to dry eyes
and dry mouth. Dr. Amulya Giridasappa, The Oxford College of Engineering
12. Ankylosing Spondylitis:
• Affected Tissues: Spine and sacroiliac joints
• Characteristics: Inflammatory arthritis primarily affecting the spine and causing stiffness and pain.
13. Autoimmune Hemolytic Anemia:
• Affected Tissues: Red blood cells
• Characteristics: Immune system attacks and destroys red blood cells, leading to anemia.
14. Vitiligo:
• Affected Tissues: Skin
• Characteristics: Loss of pigmentation in the skin due to the immune system attacking melanocytes.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Autoimmunity

• Paul Ehrlich, the German Immunologist at the turn of the 20th Century coined a term ―horror autotoxicus‖ and
described that the immune system could do mistake and, instead of reacting against foreign antigens, could focus
its attack on self-antigens.
• Nowadays this term ―horror autotoxicus‖ is called as autoimmunity. During the development of immune system,
every individual show an important phenomena called immune tolerance. During immune tolerance the
individual‘s self reacting clones of lymphocytes undergo apoptosis without selection and proliferation. These
clones of lymphocytes which react to our own body molecules (self antigens) are selected and deleted from the
body. Thus an individual‘s immune system doesn‘t produce any immune response to his own body cells or
molecules.
• The immune system must effectively defend against diverse types of pathogenic microbes while simultaneously
maintaining tolerance to self-antigens. But under certain circumstances immune system fails to produce immune
tolerance. This results in an inappropriate response of the immune system against self-components. This
phenomenon is known as autoimmunity.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• Autoimmunity is defined as an immune response produced against self antigen, (i.e., any molecule that is a
normal body constituent of the individual) leading to destruction or impaired function of a cell, tissue or organ.
Such diseases affect 5%–7% of the human population, often causing chronic debilitating illnesses.
• Autoimmune diseases can be classified based on the following criteria
✓ Extent of organ involvement
✓ Innate immune system requirements, and
✓ Effector mechanisms involved
• However, each type of autoimmune disease has unique pathophysiologic characteristics. Based on the organ
involvement, autoimmune disorder can be classified into
• i) Organ-specific and ii) Systemic autoimmune disease
Organ-specific autoimmune disease - In an organ-specific autoimmune disease, the immune response is directed
to target antigen, that is unique to a single organ or gland, so that the manifestations are largely limited to that organ.
• The cells of the target organs may be damaged directly by humoral or cell- mediated effector mechanisms or in
some organ specific autoimmunity the antibodies may over-stimulate or block the normal function of the target
organ. Dr. Amulya Giridasappa, The Oxford College of Engineering
• Some organ specific autoimmune diseases mediated by direct cellular damage.
• Autoimmune diseases involving direct cellular damage occur when lymphocytes or antibodies bind to cell-
membrane antigens, causing cellular lysis and/or an inflammatory response in the affected organ.
• Gradually, the damaged cellular structure is replaced by connective tissue (scar tissue), and the function of the
organ declines. Eg., Hashimoto thyroiditis, autoimmune anemias, Goodpasture‘s syndrome, insulin-dependent
diabetes mellitus, Addison’s disease, vitiligo, etc.

• It is noninfectious form of inflammation of the thyroid

gland.
• Its onset is insidious, with gradual enlargement of the
thyroid gland (called goitre) and gradual decrease in
thyroid hormone production.
• The natural course of the disease often includes
gradually increasing thyroid enlargement, as well as
increasing thyroid deficiency.
• Patients with Hashimoto thyroiditis have antibodies
against several components of thyroid tissue in their
Dr. Amulya Giridasappa, The Oxford
serum.
• Among them are antibodies against the enzyme thyroid peroxidase and the unique thyroid protein
thyroglobulin.
• These antibodies are often measured as a diagnostic test for the disease, but they do not alter thyroid
function or damage the thyroid. Some patients with the disease produce antibodies that block the action of
thyrotropin (thyroid-stimulating hormone, TSH), the anterior pituitary hormone that maintains normal thyroid
function.
• In most patients the thyroid is gradually destroyed, either by antibodies that damage the thyroid cells
(cytotoxic antibodies) or, more likely, by lymphocytes that have been sensitized to migrate to the thyroid,
where they interact with thyroid cells and produce substances (cytokines) that damage thyroid cells.
• The factors that initiate this process or the particular substances that cause the thyroid damage are not
known.
• There is no treatment for Hashimoto disease itself. Hypothyroidism is treated with thyroid hormone, usually
thyroxine, which may also reduce the size of a goitre, if present.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Some other autoimmune diseases are mediated by stimulating or blocking auto-antibodies.
• In case of Graves‘ disease by stimulating auto-antibodies are produced. These antibodies act as agonists, binding
to hormone receptors instead of the normal ligand and stimulating inappropriate activity. This usually leads to an
overproduction of mediators or an increase in cell growth.
• The production of thyroid hormones is carefully regulated by thyroid-stimulating hormone (TSH), which is
produced by the pituitary gland. Binding of TSH to a receptor on thyroid cells activates adenylate cyclase and
stimulates the synthesis of two thyroid hormones, thyroxine and triiodothyronine.
• A patient with Graves‘ disease produces auto-antibodies that bind the receptor for TSH and mimic the normal
action of TSH, activating adenylate cyclase and resulting in production of the thyroid hormones. Unlike TSH,
however, the autoantibodies are not regulated, and consequently they overstimulate the thyroid. For this reason
these auto-antibodies are called long-acting thyroid-stimulating (LATS) antibodies.
• In case of Myasthenia gravis, auto-antibodies may act as antagonists, these antibodies binds to acetylcholine
receptor, binding of these antibodies to receptors causes the blocking of receptor function. This generally causes
impaired secretion of mediators and gradual atrophy of the affected organ. Eg., Myasthenia gravis

Dr. Amulya Giridasappa, The Oxford College of Engineering

Systemic Autoimmune Diseases
• Systemic autoimmune diseases are a broad range of related diseases characterized by dysregulation of
immune system which give rise to activation of immune cells to attack autoantigens which results in
inappropriate inflammation and multi tissue damages.
• In systemic autoimmune diseases, the immune response is directed toward a broad range of target antigens and
involves a number of organs and tissues. These diseases reflect a general defect in immune regulation that results
in hyperactive T cells and B cells.
• Tissue damage is widespread, both from cell-mediated immune responses and from direct cellular damage
caused by auto-antibodies or by accumulation of immune complexes.
• Eg., Systemic Lupus Erythematosus attacks many tissues, Multiple Sclerosis attacks the central nervous system,
Rheumatoid Arthritis attacks joints.
• The mechanism of pathogenesis of systemic autoimmune diseases is still not very clear. It is now considered that
genetic factors, infection, endocrine, and environmental exposure are involved in the pathogenesis of these
diseases.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Dr. Amulya Giridasappa, The Oxford College of Engineering
• A variety of mechanisms have been proposed in order explain the production of immune response during
autoimmune diseases. Evidence exists for each of these mechanisms, and it is likely that autoimmunity does not
develop from a single event but rather from a number of different events.
• Normal thymic selection appears to generate some self-reactive TH cells; abnormalities in this process may
generate even more self-reactive TH cells. Activation of these self-reactive T cells in various ways, as well as
polyclonal activation of B cells, is thought to induce an systemic autoimmune response, in this case
resulting in tissue damage.
• The induction of self-tolerance in T cells resulting from exposure of immature thymocytes to self- antigens and
the subsequent clonal deletion of those that are self-reactive. Any tissue antigens that are sequestered from the
circulation, and are therefore not seen by the developing T cells in the thymus, will not induce self-tolerance.
• Exposure of mature T cells to such normally sequestered antigens at a later time might result in their activation.
Examples for these type of sequestered antigens are sperm antigens and lens proteins of eye.
• Molecular mimicry - Numbers of viruses and bacteria have been shown to possess antigenic determinants that
are identical or similar to normal host-cell components. These antigenic determinants induce the production of
cross reacting antibodies. These antibodies may damage the organ or tissue. Example – Rheumatic fever. In this
case, the antibodies are produced against Streptococcal antigens, but they cross-react with the heart muscle.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• Rheumatoid Arthritis – It is a common autoimmune disorder most often affecting women from 40 to 60 years
old. The major symptoms of RA are - chronic inflammation of the joints, although the hematologic,
cardiovascular, and respiratory systems are also frequently affected.
• Many individuals with rheumatoid arthritis produce a group of auto-antibodies called rheumatoid factors which
react with certain antigenic determinants in the Fc region of IgG. Although, the classic rheumatoid factor is an
IgM antibody with that reactivity.
• These auto-antibodies bind to normal circulating IgG, forming IgM-IgG complexes that are deposited in the
joints. These immune complexes can activate the complement cascade, resulting in a type III hypersensitive
reaction, which leads to chronic inflammation of the joints.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Some other systemic autoimmune diseases
• The pancreatic beta cells of individuals with insulin-dependent diabetes mellitus (IDDM) express high levels of
both class I and class II MHC molecules, whereas healthy beta cells express lower levels of class I and do not
express class II MHCs at all.
• Similarly, thyroid acinar cells from those with Graves‘ disease have been shown to express class II MHC
molecules on their membranes. This inappropriate expression of MHC molecules may cause activation of TH cells
allowing activation of B cells or TC cells or sensitization of TH1 cells against self-antigens.
• A number of viruses and bacteria can induce nonspecific polyclonal B-cell activation. Gram- negative bacteria,
cytomegalovirus, and Epstein-Barr virus (EBV) causes nonspecific polyclonal activation of B cells that express IgM
in the absence of TH cells. If B cells reactive to self- antigens are activated by this mechanism, auto-antibodies
can appear. Example - infectious mononucleosis, which is caused by EB.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Animal Models for Autoimmune Diseases
• The present knowledge about the immune system and autoimmunity has been derived from studies in animals,
particularly the mouse, which has an immune system and genome composition similar to that of humans. Many
well-characterized autoimmune models exist in which investigators can manipulate genomes and immune
systems, test interventions, and modify environments.
• Animal models of autoimmune disease can be divided into three main types based on their derivation:
(1) Spontaneous
(2) Genetically modified and
(3) Experimentally induced

• Spontaneous animal models- A number of autoimmune diseases that develop spontaneously in certain
animals exhibit important clinical and pathologic similarities to certain autoimmune diseases in humans. Certain
inbred mouse strains are particularly valuable models for studying the immunologic defects involved in the
development of autoimmunity.

Dr. Amulya Giridasappa, The Oxford College of Engineering

✓ New Zealand Black (NZB) mice and F1 hybrids of NZB and New Zealand White (NZW) mice spontaneously
develop autoimmune diseases that closely resemble systemic lupus erythematosus (SLE).
✓ The SKG arthritis model is a spontaneous, inflammatory, and erosive arthritis caused by a ZAP70 mutation
that, similar to RA, is associated with rheumatoid factor (RF) and antibodies to citrullinated proteins.
✓ T1DM develops in non-obese diabetic (NOD) mice and biobreeding (BB) rats as a result of T cell–mediated
destruction of β-islet cell
Genetically modified group
• Transgenic, site-directed genetic replacement (gene knockout or knockin) and N-ethyl N- nitrosourea
(ENU)-mutagenized mice are used for the study of lupus and organ-specific diseases, particularly type 1
Diabetes millitus (T1DM) and multiple sclerosis (MS).
• The lupus models, primarily single gene knockout or transgenic mice, have provided information related to
immune tolerance and disease pathogenesis.
• Genetically modified mouse K/BxN and B6 × NOD hybrid expressing a transgenic T cell receptor, named
KRN, that recognizes a bovine ribonuclease peptide on H-2K are used as arthritis model.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Experimentally induced models
• These models are used for the study of both systemic and organ-specific diseases. More commonly studied
models of systemic disease include tetramethylpentadecane (TMPD; also called pristane)-induced
autoimmunity, mercury-induced autoimmunity, and chronic graft- versus-host disease for the study of
human SLE.
• For the induced models of organ specific diseases, a common approach is to immunize rodents with a self-
antigen or closely related peptide or foreign antigen, plus a strong adjuvant, usually complete Freund‘s.
This approach makes it possible to induce autoimmunity in virtually all organ systems and to produce
diseases mediated by cellular as well as humoral mechanisms.
• Some of the more commonly studied organ-specific models developed by this approach include collagen-
induced arthritis (CIA), proteoglycan-induced arthritis (PGIA), and experimental autoimmune
encephalomyelitis (EAE)

Dr. Amulya Giridasappa, The Oxford College of Engineering

Treatment to autoimmune Disorders
The treatment for autoimmune disorders aims to manage symptoms, control inflammation, and modulate the
immune response. The specific approach may vary depending on the type of autoimmune disorder and the organs
involved. Here are some general strategies and therapeutic options commonly used in the treatment of autoimmune
disorders:
1.Immunosuppressive Medications:
• Corticosteroids: Such as prednisone, which have potent anti-inflammatory effects.
• Disease-Modifying Anti-Rheumatic Drugs (DMARDs): Examples include methotrexate, azathioprine, and
hydroxychloroquine. These drugs can help modulate the immune system and slow disease progression in
conditions like rheumatoid arthritis.
2.Biologic Therapies:
• TNF-alpha Inhibitors: Drugs like infliximab, adalimumab, and etanercept target tumor necrosis factor-alpha and
are used in conditions such as rheumatoid arthritis and inflammatory bowel disease.
• Interleukin Inhibitors: Targeting specific interleukins (ILs), such as IL-6 (tocilizumab) or IL-17 (secukinumab), is
used in diseases like rheumatoid arthritis and psoriasis.
• B Cell Depletion: Rituximab is a monoclonal antibody that depletes B cells and is used in conditions like
rheumatoid arthritis and certain types of vasculitis. Dr. Amulya Giridasappa, The Oxford College of Engineering
3. Immunomodulators:
• Cyclosporine and Tacrolimus: These drugs suppress the immune system and are used in conditions
like psoriasis and rheumatoid arthritis.
• Mycophenolate Mofetil: Used to reduce immune activity and prevent organ rejection after
transplantation. It may also be used in certain autoimmune disorders.
4.Monoclonal Antibodies:
• Infliximab, Adalimumab, Golimumab: Targeting TNF-alpha, these drugs are used in various
autoimmune disorders.
• Rituximab: Targets CD20 on B cells and is used in diseases like rheumatoid arthritis and certain forms of
vasculitis.
5.Cytokine Inhibitors:
• Anakinra: An IL-1 receptor antagonist used in conditions like rheumatoid arthritis.
• Tocilizumab: An IL-6 receptor antagonist used in conditions like rheumatoid arthritis and systemic
juvenile idiopathic arthritis.

Dr. Amulya Giridasappa, The Oxford College of Engineering

6. Plasma Exchange (Plasmapheresis):
• Involves removing and replacing blood plasma to remove harmful antibodies or immune complexes.
Used in certain autoimmune disorders with severe symptoms.
7. Corticosteroid Injections:
• Local injections of corticosteroids into affected joints or tissues can help reduce inflammation and
alleviate symptoms.
8. Physical Therapy:
• Physical therapy and exercise programs may be recommended to improve joint function and reduce
pain in conditions affecting the musculoskeletal system.
9. Supportive Therapies:
• Supportive care, including pain management, nutritional support, and counseling, may be important
components of overall treatment.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Immune response to infection
• Infection is a phenomenon in which invasion and multiplication of pathogenic microorganisms such as bacteria,
virus and parasites occur in the body of an individual.
• An infection may be subclinical or clinical. A subclinical infection do not produce any symptoms. Whereas,
symptoms are very common in clinically apparent infection.
• An infection may remain localized affecting a specific tissue or organ in which invasion of the pathogenic
organism occurs. Whereas some infection may be systemic where infection may spread through the blood or
lymphatic vessel and affect many tissue or organs.
• In order to establish infection in susceptible host, a series of coordinated events must be shown by the
pathogenic microorganisms to avoid both innate and adaptive immunity.
• Generally, pathogens use a variety of strategies to escape destruction by the adaptive immune system.
➢ Many pathogens reduce their own antigenicity either by growing within host cells, where they are
sequestered from immune attack, or by shedding their membrane antigens.
➢ Other pathogens hide themselves by mimicking the surfaces of host cells, either by expressing molecules
with amino acid sequences similar to those of host cell-membrane molecules or by acquiring a covering of
host membrane molecules. Dr. Amulya Giridasappa, The Oxford College of Engineering
 ➢ Some pathogens are able to suppress the immune response selectively or to regulate it so that a branch of
the immune system is activated that is ineffective against the pathogen. Continual variation in surface
antigens is another strategy that enables a pathogen to escape from the immune system. This antigenic
variation may be due to the gradual accumulation of mutations, or it may involve an abrupt change in
surface antigens.
Viral Infections
• Viral infection is the proliferation of the pathogenic virus in the body. The fundamental process of viral infection is
the expression of the viral replicative cycle in a host cell. A number of specific immune effector mechanisms,
together with nonspecific defense mechanisms, are produced by the host immune system to eliminate an
infecting virus.
• Specific immune effector mechanism produced during viral infection involves humoral and cell mediated immune
response. The nonspecific defense mechanism is mediated by interferons and NK cells.
• Innate immunity to viral infection - The innate immune response to viral infection is produced by the
generation of type I interferons (IFN-α and IFN-β) and the activation of NK cells.
➢ Double stranded RNA (dsRNA) produced during the viral life cycle can induce the expression of IFN-α and
IFN-β by the infected cell.
Dr. Amulya Giridasappa, The Oxford College of Engineering
 ➢ Macrophages, monocytes, and fibroblasts also are capable of synthesizing these cytokines during the viral
infection.
➢ IFN -α and IFN-β can induce an antiviral response or resistance to viral replication by binding to the IFN α/β
receptor. Once bound, IFN –α and IFN-β activates the JAK-STAT pathway, which in turn induces the
transcription of several genes. One of these genes encodes an enzyme known as 2-‫׳‬5-‫׳‬oligo- adenylate
synthetase [2-5(A) synthetase], which activates a ribonuclease (RNAse L) that degrades viral RNA.
➢ Other genes activated by IFN-α/β binding to its receptor also contribute to the inhibition of viral replication. For
example, IFN- α/β binding induces a specific protein kinase called dsRNA-dependent protein kinase (PKR),
which inactivates protein synthesis, thus blocking viral replication in infected cells.
➢ The binding of IFN-α and IFN-β to NK cells induces lytic activity, making them very effective in killing virally
infected cells. The activity of NK cells is also greatly enhanced by IL-12, a cytokine that is produced very early
in a response to viral infection.
• Acquired immunity to viral infection - Specific immune effector mechanisms produced during the viral infection
are antibody mediated humoral immunity and cell mediated immunity.
➢ B Cells: Produce antibodies (immunoglobulins) that can neutralize viruses, prevent their entry into cells, and tag them for
destruction by other immune cells.
➢ T Cells: Cytotoxic T cells directly kill infected cells, while helper T cells coordinate the immune response and stimulate B cells.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
Immunity to bacteria
• Bacterial infection has enormous impact on human society, which exerts major threat to public health.
• Bacteria enter the body either through a number of natural routes (e.g., the respiratory tract, the gastrointestinal
tract, and the genitourinary tract) or through normally inaccessible routes opened up by breaks in mucous
membranes or skin.
• Type of immune response produced depends on the number of organisms entering and their virulence. If the
inoculum size and if the virulence of the pathogenic bacteria are low, then nonspecific innate immunity involving
localized tissue phagocytes will eliminate the invading pathogenic bacteria. Larger inoculums or organisms with
greater virulence tend to induce an adaptive, specific immune response.
• Humoral Immune Responses to Extracellular bacteria - The extracellular bacteria are capable of replicating
outside the host cells such as in the circulation, connective tissues, the tissue space, the lumen of airways and
gut. They may induce localized inflammatory response and tissue destruction, pus formation.
• Some of them produce toxins (endotoxin or exotoxon). The endotoxins of Gram negative bacteria (LPS) are a
potent activator of macrophages. Exotoxin, can be cytotoxic but also may cause pathogenesis in other ways.
• The innate immunity and humoral immune response is the main protective response against extracellular
bacteria.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Innate immunity to extracellular bacteria
• Skin and exposed epithelial surfaces have nonspecific innate protective system. Skin prevents entry of
pathogens; it prevents growth of bacterium due to their relative dryness (keratin) and by the effect of sebum.
• Commensals can limit bacterial evasion by producing antibacterial protein called colicin.
• Peptidoglycan of Gram positive bacteria and LPS on the cell walls of Gram negative bacteria leads to
activation of complement system by alternative pathway which leads to lysis of bacterial membrane by MAC.
The pathway also produces opsonins (C3b) which helps in phagocytosis of bacteria by opsonization.
• Many cells such as phagocytes, dendritic cells, epithelial cells and B-cells have certain non specific receptors
called Toll-like receptors (TLRs). These TLRs recognize non specific antigens present on the bacteria called
pathogen-associated molecular patterns (PAMPs). These toll like receptors helps in phagocytosis of the
antigens, activation of T cells and induce inflammatory response.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Acquired Immunity to extracellular bacteria - Humoral immunity is the major type of specific immune
response produced by immune system against the extracellular bacteria.
i. Antibodies (IgG and IgA) produced by plasma cells bind to epitopes on the surface of toxins and neutralize
their action by preventing its binding to host cell surface. The antibody-toxin complexes are then cleared by
phagocytic cells in the same manner as any other antigen-antibody complex.
ii. Complement activation by classical pathway (by IgG and IgM) can lead to lysis of the organism by MAC.
iii. Antibody (IgG) that binds to accessible antigens on the surface of a bacterium and C3b component of
complement, can act as an opsonin that increases phagocytosis and thus clearance of the bacterium.
iv. The complement split products C3a, C4a, and C5a act as anaphylatoxins, inducing local mast-cell
degranulation and thus vasodilation and the extravasation of lymphocytes and neutrophils from the blood into
tissue space.
v. Other complement split products such as C5a, C3a, C5b67 serve as chemotactic factors for neutrophils and
macrophages, thereby contributing to the buildup of phagocytic cells at the site of infection.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Cell mediated immunity to extracellular bacteria - The protein antigens of extracellular bacteria activate TH
cells with the help of APC and causes following effect
• Secrets cytokines that help in activation of B-cells
• Induce local inflammation and enhance the microbicidal and phagocytic activities by neutrophils and
macrophages
• Secret IFN-γ responsible for macrophage activation which ensures phagocytosis and killing of bacteria
• Secret TNF and lymphotoxin which trigger inflammation.

Dr. Amulya Giridasappa, The Oxford College of Engineering

 Acquired Immunity to extracellular
bacteria

Dr. Amulya Giridasappa, The Oxford College of Engineering

Immunity to intracellular bacteria - When a bacterium is intracellular, it survives and replicates within the cell.
These bacteria are inaccessible to circulating antibodies. Their elimination requires the mechanism of cell
mediated immunity.
• Innate immunity is not very effective against intracellular bacterial pathogens. Intracellular bacteria can
activate NK cells, which in turn provide an early defense against these bacteria. Intracellular bacterial
infections tend to induce a cell-mediated immune response, specifically, delayed type hypersensitivity (DTH).
• During DTH, cytokines secreted by CD4+ T cells are important—especially IFN-γ, which activates
macrophages to kill ingested pathogens more effectively by the production of several antimicrobial substance
such as reactive oxygen species, nitric oxide and lysosomal enzymes.
• Phagocytosed bacteria stimulate CD8+ T cell responses if the antigens are transported from phagosomes into
the cytosol or escape from the phagosome or the phagocytes cannot destroy the bacteria. These CD8+ T
cells produce CTLs which will help in the killing of infected cells.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Immunity to fungal infection
• Fungal infections are normally only a superficial nuisance (e.g. ringworm: top), but a few fungi can cause serious
systemic disease, usually entering via the lung in the form of spores.
• The outcome depends on the degree and type of immune response, and may range from an unnoticed respiratory
episode to rapid fatal dissemination or a violent hypersensitivity reaction.
• Predominant defense mechanisms differ depending on the specific causative agent.

• Immune response to fungi consist mainly of :

1) Innate immunity is mediated by
❑ Neutrophils and macrophages
❑ Fungi are readily eliminated by phagocytes
❑Activated neutrophils are critical in the defense against disseminated candidiasis and aspergillosis
2) Acquired immunity (cell mediated immunity)
❑ CMI acts in a manner similar to its action against intracellular bacteria
• Cell-mediated immunity predominates in protection against cryptococcosis, histoplasmosis and mucosal C.
albicans infection. Dr. Amulya Giridasappa, The Oxford College of Engineering
◼ In general, the survival mechanisms of successful fungi are similar to those of bacteria: antiphagocytic capsules
(e.g. Cryptococcus), resistance to digestion within macrophages (e.g. Histoplasma, etc.), and destruction of
polymorphs (e.g. Coccidioides).
◼ Some yeasts activate complement via the alternative pathway, but it is not known if this has any effect on survival.

Immunity to protozan infection

• Protozoans are unicellular eukaryotic organisms. They are responsible for several serious diseases in humans,
including amoebiasis, Chagas‘ disease, malaria, leishmaniasis, and toxoplasmosis.
• The type of immune response that develops to protozoan infection and the effectiveness of the response partly
depends on the location of the parasite within the host.
• Many protozoans have life-cycle stages in which they are free within the bloodstream, and it is during these stages
that humoral antibody is most effective.
• Many of these same pathogens are also capable of intracellular growth; during these stages, cell-mediated
immune reactions are effective in host defense.
• Helminths are large parasites that normally do not multiply within cells.
• Immune-system exposure to helminths is limited; consequently, only a low level of immunity is induced.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• A cell-mediated response by CD4+ T cells plays a critical role in the response to Schistosoma.
• CMI in response to helminthic parasites is mediated by TH2 cells that stimulate the production of IgE and
activation of eosinophils.

Innate immunity
➢ The innate immune system involves phagocytosis and cytotoxicity mediated by NK cells.
➢ The phagocytes has Pattern Recognition Receptors (PRR) (Toll-like receptors, mannose receptors and scavenger
receptors) that recognizes Pathogen associated molecular patterns (PAMPs) and causes phagocytosis.
➢ Ficolins, collectins and pentraxins etc. act as soluble opsonin by binding to pathogen surface and enhancing their
phagocytosis by phagocytes.
Humoral immunity – here, the Abs produced by plasma cells can act directly on protozoa to damage them either by
itself of by activating the complement pathways.
➢ Antibody can neutralize the parasites directly by blocking its attachment to a new host cell. Antibody can enhance
phagocytois by macrophages, the effects are mediated by Fc and complement receptors on macrophages.
➢ Antibody eliminates the protozoans by ADCC.
Dr. Amulya Giridasappa, The Oxford College of Engineering
To sumup…..

Dr. Amulya Giridasappa, The Oxford College of Engineering

IMMUNODEFICIENCIES

It is a condition in which the immune system is unable to produce an immune response against any infectious
agent or self transformed cells. Thus it fails to protect the host from disease- causing agents or from malignant
cells. Immunodeficiency can be classified into two types:
i) Primary immunodeficiency - A condition resulting from a genetic or developmental defect in immune system is
called as primary immunodeficiency. In these conditions, the defect is present at the time of birth but it may not
manifest itself until later in life.
ii) Secondary immunodeficiency - Also called acquired immunodeficiency, it is a condition in which the loss of
immune function occurs as a result of extrinsic factors such as disorders of blood or bone marrow, metabolic
disorders, drugs, treatment of cancers and infectious agents such as HIV.

Dr. Amulya Giridasappa, The Oxford College of Engineering

i. Primary immunodeficiency - A condition resulting from a genetic or developmental defect in the immune
system is called a primary immunodeficiency. In these conditions, the defect is present at the time of birth but it
may not manifest itself until later in life. A primary immunodeficiency may affect either adaptive or innate
immune functions. It is further subdivided into-
a) Lymphoid immunodeficiency– Involves components of adaptive immunity such as T or B cells or both. Thus it
may affect humoral or cell mediated immune system or both. Example – Severe Combined Immunodeficiency
Syndrome (SCID), Wiskott-Aldrich Syndrome, Bare lymphocyte Syndrome, DiGeorge syndrome, X-linked
agammaglobulinemia.
b) Myeloid immunodeficiency - defects in the myeloid cell lineage affect the innate immune functions. Most of
these defects result in impaired phagocytic processes that are manifested by recurrent microbial infection of
greater or lesser severity. There are several stages at which the phagocytic processes may be faulty; these
include cell motility, adherence to and phagocytosis of organisms, and killing by macrophages. These include
diseases like congenital agranulocytosis, chronic granulomatous disease and Leukocyte- adhesion deficiency.
c) Immunodeficiency/Immune complex disease – They arise due to defects in the complement system. Many
complement deficiencies are associated with increased susceptibility to bacterial infections and/or immune-
complex diseases. Dr. Amulya Giridasappa, The Oxford College of Engineering
• The consequences of primary immunodeficiency depend on the number and type of immune system
components which are involved.
• Defects in components early in the hematopoietic developmental scheme affect the entire immune system. In
this category lies the reticular dysgenesis, a stem-cell defect that affects the maturation of all leukocytes; the
resulting general failure of immunity leads to susceptibility to infection by a variety of microorganisms.
• Without aggressive treatment, the affected individual usually dies young from severe infection. In the more
restricted case of defective phagocytic function, the major consequence is susceptibility to bacterial infection.
• Defects in more highly differentiated compartments of the immune system have consequences that are more
specific and usually less severe. For example, an individual with selective IgA deficiency may enjoy a full life
span, troubled only by a greater than normal susceptibility to infections of the respiratory and genitourinary
tracts.
• In next figure shows congenital defects that interrupt hematopoiesis or impair functioning of immune-system
cells. These defects result in various immunodeficiency diseases.

Dr. Amulya Giridasappa, The Oxford College of Engineering

 (Orange boxes- phagocytic deficiencies,
green - humoral deficiencies, red- cell
mediated deficiencies, and
purple- combined immunodeficiencies,
defects that affect more than one cell
lineage.)

Dr. Amulya Giridasappa, The Oxford College of Engineering

ii. Secondary immunodeficiency Also called acquired immunodeficiency. It is a condition in which the loss of
immune function occurs as a result of extrinsic factors like disorders of blood or bone marrow, metabolic disorders,
drugs, treatment of cancers and infectious agents such as HIV.
• AIDS-Acquired Immunodeficiency syndrome is a type of acquired immunodeficiency that results from an infection
caused by a retrovirus, Human Immunodeficiency Virus I (HIV-I).
• HIV-1 infection is spread mainly by sexual contact, passage of blood, and from HIV-infected mother to infant.
Infection with HIV-1 results in severe impairment of immune function marked by depletion of CD4+ T cells and
death from opportunistic infection, usually within 10 years of infection.
Structure of HIV - Each virion expresses 72 glycoprotein projections composed of gp120 and gp41.
• The gp41 molecule is a transmembrane molecule that crosses the lipid bilayer of the viral envelope. Gp120 is
associated with gp41 and serves as the viral receptor for CD4+ on host cells.
• The viral envelope is derived from the host cell and contains some host-cell membrane proteins, including class I
and class II MHC molecules.
• Within the envelope is the viral core, or nucleocapsid, which includes a layer of a protein called p17 and an inner
layer of a protein, called p24.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• The HIV genome consists of two copies of single-stranded RNA, which are associated with two molecules of
reverse transcriptase (p64), Nucleoid proteins p10 which is a protease and p32 an integrase.

• HIV infection of target cells – a). HIV gp120 binds to CD4+ on target cell. Fusogenic domain in gp41 and
CXCR4, a G-protein–linked receptor in the target-cell membrane, mediate fusion.
Dr. Amulya Giridasappa, The Oxford College of Engineering
b) Nucleocapsid containing viral genome and enzymes enters cells.
c) Viral genome and enzymes are released following removal of core proteins.
d) Viral reverse transcriptase catalyzes reverse transcription of ssRNA, forming RNA–DNA hybrids.
e) Original RNA template is partially degraded by ribonuclease H, followed by synthesis of second DNA strand to
yield HIV dsDNA.
f) The viral dsDNA is then translocated to the nucleus and integrated into the host chromosomal DNA by the viral
integrase enzyme p32.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Activation of provirus - Transcription factors stimulate transcription of proviral DNA into genomic ssRNA and, after
processing, several mRNAs.
• Viral RNA is exported to cytoplasm. Host-cell ribosomes catalyze synthesis of viral precursor proteins.
• Viral protease cleaves precursors into viral proteins.
• HIV ssRNA and proteins assemble beneath the host-cell membrane, into which gp41 and gp120 are inserted.
• The membrane buds out, forming the viral envelope.
• Released viral particles complete maturation; incorporated precursor proteins are cleaved by viral protease
present in viral particles.

Injury and inflammation

• Injury and inflammation are closely linked physiological responses that the body employs to protect itself, repair
damage, and maintain tissue homeostasis. Inflammation is a complex biological response triggered by harmful
stimuli, such as pathogens, tissue injury, or irritants.
• The process involves a coordinated series of events mediated by cells, blood vessels, and soluble mediators.
• The inflammatory process is tightly regulated, involving a balance between pro-inflammatory and anti-
inflammatory signals. Dr. Amulya Giridasappa, The Oxford College of Engineering
• While acute inflammation is a normal and beneficial response, chronic inflammation can contribute to various
diseases.
• Understanding the mechanisms of inflammation is essential for developing therapeutic strategies to modulate
immune responses and promote tissue repair.
Below events indicates how injury is related to inflammation-
1. Initiation of Inflammation: Tissue injury can result from various causes, including physical trauma, infections,
or chemical exposure. When cells are damaged, they release signaling molecules, known as damage-associated
molecular patterns (DAMPs), that activate the immune system.
2. Vascular Changes:
•Vasodilation: Blood vessels near the site of injury dilate to increase blood flow, allowing immune cells and
nutrients to reach the affected area.
•Increased Permeability: The permeability of blood vessel walls increases, facilitating the movement of immune
cells, proteins, and fluids from the bloodstream to the injured tissue.
3. Immune Cell Recruitment: In response to chemical signals, white blood cells, particularly neutrophils and later
monocytes, migrate to the site of injury. Immune cells engulf and digest debris, pathogens, and damaged cells to
eliminate threats.
Dr. Amulya Giridasappa, The Oxford College of Engineering
4. Cytokine Release: Immune cells release cytokines, such as tumor necrosis factor (TNF), interleukins (ILs), and
chemokines, to regulate inflammation and immune responses.
5. Tissue Repair and Regeneration:
•Fibroplasia: Fibroblasts proliferate and produce collagen to repair damaged tissue.
•Angiogenesis: New blood vessels form to support tissue repair and deliver oxygen and nutrients.
6. Chronic Inflammation: In some cases, inflammation may persist, leading to chronic inflammation. This can
contribute to various diseases, including rheumatoid arthritis, atherosclerosis, and inflammatory bowel diseases.
7. Systemic Effects includes acute-Phase Response here inflammation can trigger systemic responses, such as
fever, increased production of acute-phase proteins (e.g., C-reactive protein), and changes in metabolism.
8. Pain and Sensitization: - Nociceptor Activation: Inflammation may stimulate pain receptors (nociceptors),
contributing to pain and sensitivity in the affected area.
VACCINES
• Prevention of disease is the most desirable, convenient and highly effective approach to a good health. The
disease can be prevented by vaccination or immunization using biological preparations called vaccines.
Vaccines were used by Edward Jenner in 1796 in order to stimulate immunity against the small pox virus.
• The term ‗vaccine‘ is derived from the Latin term ‗vacca‘ which means ‗cow‘.
Dr. Amulya Giridasappa, The Oxford College of Engineering
• Vaccines are antigenic preparations which administered to an individual intentionally to provide immunity against
a disease.
• The process of introduction of vaccine into an individual to provide protection against a disease is called
vaccination.
• Vaccines may be composed of attenuated or dead causative agents or products of their life activity.
• These antigens generate the primary immune response, and the memory В and T cells.
• When the vaccinated person is attacked by the same pathogen, the existing memory T or В cells recognize the
antigen quickly and attack the invaders with a massive production of lymphocytes and antibodies.
• Properties of an ideal vaccine – An ideal vaccine should have the following characteristic features:
➢ It should be safe - it should not be tumorogenic, toxic or pathogenic in nature.
➢ It should have very low levels of side effects, e.g., fever after immunization etc., in normal individuals.
➢ It should not cause problems in individuals with an impaired immune system.
➢ It should not contaminate the environment.
➢ It should be effective in producing long-lasting humoral and cellular immunities.
➢ The technique of vaccination should be simple.
➢ The vaccine should be cheap so that it is generally affordable.
Dr. Amulya Giridasappa, The Oxford College of Engineering
1. Conventional vaccines
• These consist of whole pathogenic organisms which may either be killed or live microorganisms. These are
further sub classified into –
i. Live vaccines and ii. inactivated vaccines
i. Live vaccines are also called as attenuated vaccines. These vaccines are composed of whole pathogenic
agents whose virulence has lost or reduced. Attenuation of the virus may be achieved by passage of the virus
through a non specific host. Such as – tissue culture, embryonated eggs, live animals. e.g., most live vaccines are
viral vaccines - Sabin vaccines for polio, measles vaccines, mumps vaccines, rubella vaccine, chicken pox
vaccines, rota virus vaccines
ii. Inactivated vaccines are also called as killed vaccines. These vaccines are composed of whole pathogenic
microbes which are killed by physical or chemical method. Physical method – Heating, Chemical method –
Formalin or phenol treatment e.g., most inactivated vaccines are composed of bacterial vaccines and some viral
vaccines – Salk vaccines for polio, typhoid vaccine, cholera vaccine etc.

Dr. Amulya Giridasappa, The Oxford College of Engineering

The conventional vaccines have several drawbacks:
• It is not possible to develop vaccines for the organisms which do not grow in culture
• The yield of vaccines is very low
• Cell cultures are costly to maintain
• There is a danger of non-virulent organisms getting converted to virulent ones. Vaccinations by such organisms
may cause the disease itself
• It is not possible to prevent all the disease by use of traditional vaccines e.g., AIDS.

2. Purified antigen vaccines These are based on purified antigens isolated from concerned pathogens. Since
these vaccine preparations do not contain organism, therefore the pathogenicity is avoided. These includes-
i) Purified polysaccharide antigens- vaccines based on polysaccharide antigens from the bacterial cell wall
capsules of N. meningitis, S. pneumonia. These antigens will induce humoral immune response in the host.
ii) Toxoid vaccines - Toxoids are inactivated toxins. These are vaccines directed at the toxins produced by a
pathogen. Such as vaccines against tetanus and diphtheria.

Dr. Amulya Giridasappa, The Oxford College of Engineering

iii) Conjugated vaccines - developed in recent years to deal with the poor immune response of children against
certain antigens. The polysaccharide antigens isolated from the pathogenic organisms and are combined with
proteins such as diphtheria toxoid. This approach has led to the very successful vaccine for Haemophilic influenza
type B.
iv) Protein based subunit vaccines – Consists of antigenic subunits of proteins isolated from the pathogenic
organisms. Such as Acellular pertusis vaccines, Disadvantages of purified antigen vaccines are:
• However the cost of these vaccines is high due to the steps involved in purification and vaccine preparation
• Many of isolated antigens are poorly immunogenic.

3. Recombinant vaccines
• These vaccines are produced by using recombinant DNA technology.
• These vaccines are composed of either a protein or a gene encoding a protein of pathogen origin that is
immunogenic and critical to the functioning of pathogenic organism.
• Subunit vaccines – the vaccines based on recombinant proteins. Proteins are generally immunogenic and many
of them are critical for the pathogenic organisms. The genes encoding such antigenic proteins from a pathogen
can be identified, isolated and expressed in E. coli or some other suitable host for mass production of proteins.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Recombinant protein vaccines such as Hepatitis B vaccine production

Dr. Amulya Giridasappa, The Oxford College of Engineering

4. DNA vaccines
• These are the vaccines based on DNA.
• The gene encoding the required immunogenic protein is isolated, cloned and then integrated into a suitable
expression vector.
• This preparation is introduced into the individual to be immunized.
• The gene is ultimately expressed in the vaccinated individual and the immunogenic protein is expressed in
sufficient quantities to produce both humoral and cell mediated immunities.
• Approaches used –
✓ Injection of pure DNA (or) RNA preparation into muscle, use of vectors for delivery of gene (such as Vaccinia
viruses, Adenoviruses, Retroviruses, E. coli, S. typhimurium, herpes virus etc.),
✓ reimplantation of autologous cells into which the gene has been transferred and
✓ particle gun deliver of plasmid DNA which contains the gene in an expression cassette.

Dr. Amulya Giridasappa, The Oxford College of Engineering

Dr. Amulya Giridasappa, The Oxford College of Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
Production of recombinant vaccine-
• The production of recombinant vaccines involves the use of genetic engineering techniques to introduce specific
genes into a host organism, typically microorganisms such as bacteria or yeast.
• These genetically modified organisms then produce proteins or antigens that are components of the target
pathogen. Recombinant vaccines are often safer and more scalable than traditional methods that involve
growing and inactivating or attenuating the actual pathogen.
• The general steps involved in the production of recombinant vaccines are:
1.Identification of Antigens:
• Researchers identify specific antigens or proteins from the target pathogen that can induce an immune
response and confer protection.These antigens are often selected based on their ability to stimulate an
effective immune response while avoiding side effects.
2.Gene Cloning:
• The genes encoding the selected antigens are isolated and cloned. This involves inserting the target genes
into a plasmid or another vector, a small piece of genetic material that can be used to transfer the genes
into host cells.

Dr. Amulya Giridasappa, The Oxford College of Engineering

3. Transformation of Host Organism: The recombinant plasmid containing the target genes is introduced into a
host organism, which is often a microorganism such as bacteria (e.g., Escherichia coli) or yeast (e.g.,
Saccharomyces cerevisiae). This step is typically achieved using transformation techniques, where the host cells
take up the foreign DNA.
4. Expression of Recombinant Protein: The host organism begins to produce the recombinant protein encoded by
the inserted genes. The introduced genes are transcribed into messenger RNA (mRNA), and the host's cellular
machinery translates this mRNA into the target protein.
5. Purification of Recombinant Protein: The recombinant protein is extracted and purified from the host cells.
Various purification techniques, such as chromatography, filtration, and centrifugation, are used to isolate and refine
the target protein.
6. Formulation of the Vaccine: purified recombinant protein is combined with other components to create the final
vaccine formulation. These components may include adjuvants (substances that enhance the immune response),
stabilizers, and preservatives.
7. Quality Control: Rigorous quality control measures are implemented to ensure the safety, efficacy, and
consistency of the vaccine. Testing includes assessments of the purity of the recombinant protein, absence of
contaminants, and overall vaccine stability. The packed vaccines are then distributed.
Dr. Amulya Giridasappa, The Oxford College of Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
Dr. Amulya Giridasappa, The Oxford College of Engineering
The production of a recombinant vaccine for hepatitis B surface antigen (HBsAg)
• Subunit recombinant vaccines are components of pathogenic organisms. They are highly pure in their
preparation, stable and safe. Hepatitis B has widespread in humans and affects liver causing chronic
hepatitis, cirrhosis and liver cancer.
• Hepatitis B virus is a 42 nm particle called Dane particle. It contains of viral genome in core, surrounded by
phospholipid envelope carrying surface antigens.
• Gene encoding for hepatitis B surface antigen (HBsAg) has been identified. Recombinant hepatitis B vaccine
as a subunit vaccine, is produced by cloning HBsAg gene in yeast cells.
• Gene of HBsAg i.e pMA 56 is linked to alcohol dehydrogenase promoter. These plasmids are then
transformed and cultured.
• The transformed cells are grown in tryptophan medium, free medium and are selected and cloned.
• HBsAg gene is expressed to produce 22nm sized particles similar to those found in patients infected with
hepatitis B. The subunit HBsAg as 22nm particles can be isolated and used to immunize patients.
It involves genetic engineering techniques to introduce the gene encoding the HBsAg into a host organism, such
as yeast cells. The steps involved in the production of a recombinant hepatitis B vaccine are:

Dr. Amulya Giridasappa, The Oxford College of Engineering

1.Identification of HBsAg: The HBsAg is the surface protein of the hepatitis B virus (HBV) and is the primary target
for immune response in the vaccine. Identify the gene that codes for the HBsAg.
2.Gene Cloning: The gene encoding HBsAg is isolated and cloned into a suitable vector, often a plasmid or another
expression vector. The vector is designed to facilitate the expression of the HBsAg gene in the chosen host.
3.Transformation of Host Organism: The recombinant plasmid containing the HBsAg gene is introduced into a
host organism such as Saccharomyces cerevisiae which is capable of expressing the gene and producing the
corresponding protein.
4.Expression of Recombinant HBsAg: The host yeast cells are allowed to multiply and express the HBsAg
protein. The HBsAg gene is transcribed into mRNA, and the cellular machinery translates this mRNA into the HBsAg
protein.
5. Harvesting and Cell Disruption: The yeast cells are harvested, and the HBsAg protein is extracted. The cells
may undergo a disruption process to release the intracellular contents, including the HBsAg protein.
6. Purification of Recombinant HBsAg: They is purified from the cell extract using various techniques such as
chromatography and filtration. This help remove impurities and isolate the HBsAg protein in its pure form.
7. Formulation of the Vaccine: The purified HBsAg protein is formulated into the vaccine. Adjuvants, stabilizers,
and preservatives may be added to enhance the vaccine's effectiveness, stability, and shelf life.
Dr. Amulya Giridasappa, The Oxford College of Engineering
 • Market name – Recombivax
and Engerix-B.
• 3 doses over a period of six
months

Dr. Amulya Giridasappa, The Oxford College of Engineering