Immunity (Resistance)

Definition: Immunity is the sum of all mechanisms used by the body for
protection against environmental agents that arc foreign to the body, and
removal of damaged tissues as well as for immune surveillance.
Functions of the immune System
1-Recognition and prevention of infection
2-Removal of tissue debris (e.g., homeostasis)
3-Detection and removing altered self-structures (e.g., immune
surveillance)
Innate and Acquired Immunity
There are 2 types of resistance mechanisms:
1. Non-specific (innate or natural) resistance and
2. Specific (acquired or adaptive) resistance.
Examples of which are presented in the following table. Impairment of
these mechanisms increases an individual's susceptibility to recurrent
infections often caused by opportunistic organisms.
I -Non -specific. or innate immunity
Components of Non-specific Immunity (Natural or Innate)
A- Mechanical Barriers:
They are found at all surfaces exposed to the external environment. They
serve to prevent entry of most pathogens into the body by inhibiting the
attachment and penetration of infectious agents. They include:
1) intact skin is the 1st line of defense against infection. Any skin
abrasions; wound, bums, facilitate entry of the organism. Organisms
might gain entry through sebaceous glands and hair follicles; however,
they are protected by other factors e.g., fatty acids and lysozymes.

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2) Oral cavity, enzymes and antibodies in saliva, flow of saliva to throat,
epithelial cells and mucous layer have protective action against infectious
agents.

3) Hairs at the nostrils act as sieve.

4) Mucous membranes and mucous secretions, the gastrointestinal tract,
respiratory tract, and genitourinary tract are bound with mucous
membranes that secrete mucous. Pathogens are trapped in the viscous
mucous and removed by other mechanisms. E.g.,
• Beating of cilia on epithelial cells in respiratory tract (RT) remove
microorganisms that become trapped in the mucous.
• Coughing and sneezing dislodge and help to expel the mucous
blanket. (Alcohol, cigarette smoking and narcotics suppress these
defense mechanisms)
5) Shedding of cells that carry microbes provides a mechanical cleansing
action.
6) High flow rates of secretions assist in flushing microbes from the
body. E.g., saliva, tears , urine, perspiration, biliary tract, and lower RT.
7) Reflex vomiting, and diarrhea are also eliminate pathogenic organisms.
8) Eyeball has a tough epithelial layer that is fairly resistant to penetration
and is continuously washed with tears.
B- Chemical and biochemical barriers
They are found in blood, lympn, and secretions as a natural defense
against microorganisms that invade the body. They inactivate and kill
many pathogens.
(1) Chemical barriers:
a) The acid pH in urine, stomach, and vaginal secretions prevents
colonization by microorganisms.

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b) Organic acids found at low pH in sebaceous gland secretions inhibit
growth of microorganisms e.g., fatty acids in sebaceous glands & lactic
acid in sweat.
c) Bile acids interfere with the vital functions of the cell membrane of
bacteria,
(2) Biochemical barriers and soluble factors
a) Lactoferrin and transferrin are iron-binding proteins that compete with
bacteria for their essential metabolite
b)Enzymes:
• Lysozyme, present in body secretions (tears, saliva, nasal secretions,
serum (30mg/ml) and in phagocytic cells). They lyse bacteria by
destroying the peptidoglycan layer of Gram positive microorganisms,
making the cell susceptible to osmotic lysis. They potentiate the action of
complement on Gram negative microorganism causing bacterial lysis.
Hydrolytic enzymes in saliva
• Proteolytic enzymes in GIT (stomach and small intestine)
• DNAse & RNAse
a) Alternative complement pathway works in conjunction with the innate
system to rid the body of invading microorganisms. Endotoxin of bacteria
(LPS) and some acute phase protein activate it leading to lysis of
microorganisms.
Cytokines include: Interleukines (IL-1, 6, 8), and tumor necrosis factor
alpha (TNF a)and interferons (IFNs). They are secreted by monocytes
and macrophages. They are proinflammatory cytokines (promote
inflammation and initiate specific immune response).
• IFNs are proteins produced usually by virally infected cells and
protect other cells in the area by inhibiting viral replication. There
are 2 types of IFNs (1 & II):
Type I is involved in innate immunity, includes:
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* IFN- α , secreted by inacrophagci and leukocytes
* IFN-β secreted by fibroblasts
c) Acute phase proteins. These are enzymes and factors whose production
is induced by tissue injury, e.g., C-reactive protein (CRP). They bind to
bacterial surface activating the complement and causing lysis of bacteria.
C-Physiologic Factors:
1 . Oxygen tension, high O2 tension in the lungs, inhibits growth of
anaerobes.
2. Body temperature. The host may respond to infection by fever (i.e.,
rising body temperature above optimum temperature for growth of
pathogen) which slow the growth rate of many pathogens allowing body's
defense to eradicate the pathogen. Fever may speed up body defense
mechanisms.
3. Inflammation. Inflammation can be considered as that complex series
of events that develops when the body is injured either by mechanical or
chemical agents or by self- destructive (autoimmune) processes.
• It aims to hinder the spread of pathogen into general circulation
(decreased venous & lymphatic return).
• It is designed to deliver soluble and cellular factors to the affected area.
The inflammatory process include:
a- Vasodilatation of blood vessels and influx of leukocytes and fluids
characterize the acute inflammation. The result is redness, (erythema),
swelling (edema) and firmness (induration).
b- Activation of endothelial cells by inflammatory mediators or
proinflammatory substances secreted by activated cells, e.g., histamine,
heparin, tryptase and others. Thus become adhesive to leukocytes
initiating the process of migration.
c- Migration of the inflammatory cells to the site of tissue injury.
Leukocytes migrating into tissues responding to a variety of chemical

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attractants at the sites of infections (e.g. chemotactic factors). (I)
Neutrophils represent the 1st line of defense against invading
microorganisms. The primary function of neutrophils is to ingest
(phagocytose) and destroy potentially dangerous agents, such as bacteria,
(ii) Monocytes augment the protective barrier, monocytes augment the
defense by adding their own phagocytic function to the area, (iii)
lymphocytes convey the immunologic capacity to respond to foreign
agents by specific humoral and cell mediated phenomena.
- Tissue injury activates kinin system with production of bradykinin,
resulting in increased permeability, decreased arterial resistance, smooth
muscle contraction and pain.
Inflammation is usually accompanied by fever which is caused by;
1- Bacterial products (endotoxin of gram-negative bacteria).
2- Endogenous pyrogens (IL-1, TNF) produced by monocytes and
macrophages.
D-Microbial factors (normal microbial flora e.g., GIT): act against
pathogenic organisms by interference mechanisms such as:
* Competition for nutrients,
* Production of substances, which kill other bacteria {e.g., lactic acid,
H2O2, bacteriocein (antibiotic like substances)}.
* Also, production of natural Abs by the body against normal flora can
cross react with pathogenic organism.
E- Cellular Factors:

1-Professional phagocytic cells:

a- Polymorphoneuclear leukocytes, PMNs (neutrophils) circulate in the

blood.

• Granulocytes, constitute 50-70% of WBC.

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 • Live 1-2 days (must be continuously replaced by bone marrow).
• After they engulf and kill several microorganisms they die.
• They are the first to arrive at the site of tissue damage in high
number to control the situation. So, they usually kill more of
bacterial invaders than other cells.
• However, they are not very effective against eukaryotic pathogens
as fungi and parasites.
• They have receptors For IgG (Fc-R) and complement component
(C3b).
b- Monocytes and macrophages lineage:
• Monocytes circulate in the blood
• Macrophages are wondering in spaces and present in all body
tissues. When they become fixed in tissues, they have special
name, e.g., Kupffer's cells (liver), Langerhans 'cells (skin), alveolar
rnacrophages (lung), microglial cells (CNS), and histiocyles
(connective tissue).
1- Granulocytes other than PMN (e.g., eosinophils, basophils and
mast cells)
2- NK cells: These are large granular lymphocytes, they are naturally
cytotoxic cells.
Functions of phagocvtic cells
1- Act as effector cells in preventing infections by removing antigen and
killing viable organisms.
2- Present antigen and initiate inflammation. E.g., macrophages, dendritic
cells, Langerhan's cells of skin, B cells & Endothelial cells.
3- Generate factors to repair the damaged tissues.
4- Secrete a wide variety of biologically active compounds,
Phagocytosis

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 It is the process by which particulate substances such as bacteria
are ingested by a cell (phagocytic cell) and destroyed. The most potent
macrophage activators are lipopolysaccharides (LPS) from bacteria and
gamma interferon (INF-y) by activated T cells.

Steps involved in phagocytosis

1- Chemotaxis (attraction) and chemotactic factors. The phagocyte must
come into physical contact with target which is facilitated by the
presence of:
• Receptors molecules on phagocytic cells, which are scattered uniformly
over their surfaces, e.g., receptors for opsonins (C3b), for Fc portion of
certain immunoglobulin molecules.
• Adhesion molecules (ADM), they are glycoproteins expressed on cell
surfaces that play a mechanical role in the binding of cells to its
environment either to extracellular matrix or to another cell. They are also
involved in the transduction of powerful signals controls cell activation
and proliferation. ADM include selectins, intergrins, some of
immunoglobulin superfamily and mucin. Selectins and integrins help
phagocytes to adhere to vessel walls before they leave blood vessels
towards chemotactic factors.
• Chemotactic factors may be derived from bacteria e.g.,
lipopolysaccharide endotoxin, or the host itself e.g., C5a, IL-8.
2- Recognition, phagocytes have intrinsic ability to recognize bacteria
and damage tissues without destroying normal self-tissue. This process is
significantly enhanced by factors (i.e., opsonins) that label foreign
material for the phagocytes. This process is called opsonization enhanced
phagocytosis'. E.g., of major opsonins are C3b, IgG (IgGl and IgG3),
CRP, leukotrienes and fibronectin.

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3- Engulfment. When a phagocyte makes contact with a particle, it
engulfs the particle, surrounding it with part of its cell membrane. The
membrane enclosing the particle compresses and moves into the
cytoplasm of the cell, forming a phagocytic vacuole or phagosome.
Lysosomes, which are membrane-bound bags of enzymes, fuse with the
phagosome to form a phagolysosome.
The process of phagocytosis

4- Intracellular destruction and digestion. Inside the phagolysosome, the

engulfed material is digested by:
• Enzymes contained in lysosomal granules, which are important in
breaking down ingested material and in killing microorganisms e.g.,
lysozyme
• Generation of toxic O2 metabolites
• Nitric oxide (NO)
Other cells (granulocytes and NK cells)
- Eosinophils:
-The eosinophils are also phagocytic cells. They constitute 1-3% of
WBCs.
-The granules of the eosinophils contain histaminase, which can
inactivate products of degranulated mast cells. So, eosinophils can
suppress the reactions initiated by mast cell degranulation (type I
hypersensitivity reactions)
-They are important cells in resistance to parasites through the toxic basic
protein, which is found in the eosinophil granules.
-Eosinophils also express a cell-surface receptor for the Fc of the
antibody molecule, which allows the eosinophil to bind to antibody-

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coated parasites, releasing their major basic-protein and killing the
infecting organisms.
-An elevated eosinophil level usually indicates either an allergic response
or infection with a helminthes.
- Basophils and Mast Cells
-They constitute 0.2 % of WBCs
-Both cells are characterized by a multilobed nucleus, abundant large
cytoplasmic granules, and a unique affinity for basic dyes.
-The two cell types are distinguished from each other by their location.
Basophils are found in the circulation, and mast cells reside in the tissues.
-Both cells initiate inflammation and anaphylaxis upon stimulation by
antigen and IgE or by the activation of complement through C3a
component of complement.
-Their granules contain histamines, heparin, serotonin, TNF, leukotrienes
and prostaglangines,
- Natural killer ceils (NK cells)
They are described in cells involved in the immune responses.

Host Factors affecting Innate Immunity

1-Species. Different species have different susceptibility to infective
agents (man differs from dogs, rats', and guinea pigs).
2-Race. Different races have different susceptibility to infection, e.g.,
Negroes are more susceptible to tuberculosis, and Caucasian is more
susceptible to Diphtheria and Gonorrhea.
3-Age. Very young or very old people are more susceptible to infections
because their immune responsiveness is immature or suboptimal.
4-Nutrition. Malnutrition predisposes to infection leading to decrease
total leukocyte count and decrease phagocytic activity,

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5-Hormonal balance, e.g., Diabetes mellitus, adrenal dysfunction, and
hypothyroidism decrease resistance to infections.
II-Specific (Acquired) immunity
Specific immunity has 4 essential characteristics:
• An induction phase, which is the time interval that follows the 1st
exposure to an antigen (Ag), during which precommitted
lymphocytes proliferate and mature into Ab secreting plasma cells
or into specifically reactive T cells that will secrete various
mediators (cytokihes) following subsequent contact with the Ag.
* The ability to distinguish self from foreignness.
* Specificity, in which Abs or sensitized lymphocytes react only with
matching Ags.
* Immunologic memory, which allows Abs and sensitized lymphocytes to
remember their matching Ag and react with it later.
Specific immune responses are mediated by 2 mechanisms;
A-Humoral Immunity involves bursa or bone marrow-derived (B)
lymphocytes, or B cells. B-Cell-mediated (cellular) Immunity involves
thymus-derived (T) lymphocytes, or T cells.
*Both responses usually work together to destroy pathogenic
microorganisms or neutralize their toxic products.
Specific immunity may be acquired by natural or artificial processes.
• Examples of naturally acquired immunity include the immunity that
develops during the placental transfer of antibody (passive) and the
recovery from disease (active).
• Examples of artificially acquired immunity include administration of
antisera (passive) or vaccination (active).
Comparison between innate and acquired immunity
Innate immunity (nonspecific) Acquired immunity (specific)

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 Acts as 1st line of defense Acts as 2nd line
mechanisms
Responds immediately Takes time to act
No memory: inability to learn from Memory: remember: Ag on
experience; repeated exposure to reinfection acts faster & more
pathogen dose not speed action nor pronounced to abort infection
level of activity before disease occurs
Circulating molecules: Complement Circulating molecules: Antibodies &
(alternative) complement (classic)
Production of soluble mediators: Production of soluble mediators;
- Monokines - Cytokines
- Interferon (INF-α, β) - Interferon- γ
- Tumor necrosis factor (TNFα) - TNFβ
Specificity: No (e.g., does not depend Specificity: Yes
on specific recognition of foreign)
Components: Components: ^
- Mechanical barriers to infection - - Cell-mediated immune response
Chemical and biochemical (carried out by. T lymphocytes)
inhibitors of infection - Humoral immune response (by B
- Physiological factors lymphocytes)
- Cellular factors or immunologic
barriers (phagocytes & NKcells)
and microbial flora(competition)

Tissues and Cells Involved in Immune Responses

Tissues

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They involve lymphoid tissues in which lymphoid cells mature,
differentiate and proliferate. Tissues include lymph nodes, spleen, bone
marrow, lung and liver.
* During fetal development, precursor cells of the lymphoid system
originate in the yolk sac, liver, spleen or bone marrow. They provide
stem cells that give rise to lymphocytes, phagocytes, and non-lymphoid
cells.
* Stem cells from bone marrow or embryonic tissues are deposited in 1ry
lymphoid organs where they mature into lymphocytes.
* Lymphoid tissues are divided functionally into central (1ry) and
peripheral (2ry) lymphoid organs:
A- Central dry) Lymphoid Tissues:
I Thymus, in which T cells, mature into Ag recognizing lymphocytes.
II Bone marrow or Bursa of Fabricius, in which B cells, mature into
Ag recognizing lvmphocvtes.

B- Peripheral (2ry) Lymphoid Tissues:

They include spleen, lymph nodes, mucosal associated lymphoid
tissue or MALT (which includes tonsils, Peyer's patches, appendix,
bronchial and mammary tissue).
Functions of 2ry lymphoid tissues:
1 - Trapping and concentrating fee foreign substances.
2 - Site of production of Abs and Ag specific-T cells.
Thymus-dependent areas (T Thymus-independent areas (B cell

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cells areas) areas)

Lymph nodes: They are Lymph nodes: They are localized in

localized in paracortical area. the outer cortex, lymphoid follicles
Spleen: They are present in (germinal centers), corticomedullary
periarteriolar sheath (white junction and medullary cords
pulp). Spleen: They are present in Iry
follicles & marginal zones (white
pulp).

Cells involved in the immune response include B cells, T cells,

macrophages and NK cells.
B lymphocytes: )
- B cells constitute only 15-20 % of circulating blood lymphocytes.
- Their life span is short (days or week)
-B cells mature in the bone marrow and, on leaving the marrow, carry a
surface membrane antigen-binding receptor.
1- B-cell-development
a) In mammals, B cells originate from stem cells in the bone marrow.
b) When the B cell moves to the blood and peripheral lymphoid tissues, it
carries Ig in its surface membrane and is ready to interact with antigen.
Contact between the antigen's epitope and the Ig in the B-cell membrane
triggers cell division. This process is enhanced by interleukins secreted
by T helper (Th) cells.
c) The B cell matures into one of two types of cells, the plasma cells
(secrete large amounts of Abs) and the memory cells responsible for rapid
plasma cell proliferation in 2ry immune response.
2- B-cell surface receptors:

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 a) B-cell -antigen receptor. Immunoglobulins (chiefly monomeric
IgM and IgD), function as an Ag receptor, this initiates the
differentiation of B cell into plasma cells. b)Other markers
including:
1-Receptors of the Fc portion of Ig molecules.
2-CD antigens (CD = cluster of differentiation) are found in B cell
membranes, present at different stages of leukocytes differentiation.
These markers are detected by monoclonal Abs.
3-Receptors for complement component (e.g., CR1 & CR2). CR2 =
receptor for Epstein Barr virus (EBV). Infection with EBV frequently
produces continuous replication of line of B cells that may lead to
production of lymphoma
4- Major histocompatibility complex molecules (MHC) class I & II.
5-Receptors for costimulatory molecules (B7, CD40).
T lymphocytes:
-T lymphocyles represent 65-80 % of the circulating blood lymphocytes.
-They have long life span (months or years).
-They are thymus-dependent, responsible for CMI.
1 - T cell development: Fetal stem cells enter the thymus and proliferate.
These immature T cells (called thymocytes, as it is present in thymus)
pass through several developmental stages before leaving the thymus to
the peripheral lymphoid tissues as mature T cells.
4 Immature thymocytes differentiate into T cells by the effect of thymic
hormones.
* Thymocytes express several cell surface molecules i.e.. CD molecules, i
- Cells first acquire CD2, CD4 and CDS in the cortex,
- Then, in the medulla they acquire CD3 and then loose CD4 or CDS to
by become two populations:

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 1. CD2. CD3, CD4 (helper & delayed hypersensitiviry ceils - CD4
cells) &
2. CD2: CD3, CD4 (suppressor & cytotoxic cells = CD8 cells).
2- T cell surface markers (Receptors)
a) T-cell antigen receptors (TCRs). T cells have an antigen-specific
receptor. This TCR bears significant structural homology with the Fab
portion of Ig molecule, TCRs are heterodimers, each consisting of 2 non-
identical polypeptide chains, an a chain and a p chain, linked together by
disulfide bonds.

b) Class I and class II MBC molecules.

c) CD antigens (CD3, CD2 and CD5) are found on. most peripheral
blood T cells t
• CD3 is associated with the T cell receptor for Ag, it is important in
intracellular signaling to initiate an immune response once the cell has
interacted with a matched Ag.
• CD2 is intercellular adhesion molecule and has a role in signalling
pathway for cellular activation. CD2 is also a sheep red .blood cells
receptor responsible for resetting of sheep red blood cells in the E-rosette
assay for T cell enumeration.
• CD4 and CDS are present on different effector T cells. These receptors
.bind the MHC molecule during Ag presentation to the T cell. CD4 and
CDS play a role in activation of T cells and serve as a cell-cell adhesion
molecule and have a role in signalling transduction.
d) Fc-receptor of Ig
e) Complement-receptor
3- T Cell Subpopulations:
A-CD4+ cells [T helper(Th) cells]:

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- CD4 cells constitute 65 % of peripheral T lymphocytes.
- CD4 cells recognize Ags associated with class II MHC molecules. T
cells have the capacity to differentiate into either Th subtype. Such
differentiation probably involves an intermediary stage, designated the
Th0 cells which produces IL-2, IFN y and IL-4. Repeated stimulation in
the presence of IL-12 (macrophage-derived cytokine), causes Th cells to
differentiate into Thl cells. Stimulation in the presence of IL-4 promotes
the development of Th2 cells.

The Th subsets down-regulate one another: -

• IFN γ secreted from Thl cells suppresses the maturation of Th2 cells and
also the enhancing effect of IL-4 (produced by Th2 cells) on IgE and
IgG4 synthesis.
• IL-10 secreted from TH2 cells suppresses the production of Thl cells, it
inhibits cytokine synthesis by Thl cells. Therefore, Th2 are a type of
suppressor cell.
Thl cells mediate functions connected with cytotoxicity and local
inflammatory reaction (previously they were called T cells of delayed
hypersensitivity (TD), they secrete EL-2, IFN y, TNF (3.They promote
the development of Tc cells, activation of mactophages and production of
IgGl.
Thl cells secrete cytokines (IL4, 5, 6, and 10), they help B lymphocytes to
respond to thymus dependent antigen. IL-4 enhances class switching to
IgE, while IL-5 promotes the proliferation of eosinophilies,
A- CD8+ cells [ cytotoxic(Tc) celts].
• CDS cells constitute 35 % of peripheral T lymphocytes.
• CDS cells recognize Ag in association with class I MHC molecules.

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Cytotoxic (Tc) cells
* They cause lysis of Ag-bearing target cells.
* Tc cells rid the body of cells that express foreign or non self-Ags,
* Tc cells are induced by, and are active against, tumors, viruses, and
allogenic graft.
Tc cells can kill antigen-bearing cells by:
1- The release of cytoplasmic granules, which contain perform and
granzymes.
2- Fas ligand-Fas pathway. Antigen recognition stimulates Tc cells to
express Fas ligand, a member of the tumor necrosis factor (TNF) family.
The interaction of Fas ligand with Fas receptor (TNF-R) on the target
cellinduces apoptosis (which is a process of physiological death) in the
Fas-expressing cell.

Suppressor T cells (Ts)

-Ts cells suppress Ig production by B cells through suppression of Th
cells and delayed hypersensitivity reaction.
-Ts cells prevent the occurrence of atopy by suppressing IgE secretion.
-Ts cells may be involved in the prevention of autoimmunity (inhibit self-
reacting clones of lymphocytes).
Ts cells are functiomally but not phenotypically a distinct T cell subset.
When the body no longer needs lymphocyte stimulatory activity Th and
or Tc cells produce inhibitory cytokines as TGF-p and act as suppressor
cells. Through the same mechanism other cells can also act as suppressor
cells e.g., macrophages.
Gamma/delta (v/5 ) T cells:
• A small subset (<5%) of mature T cells
• Do not express a TCR α/β but have a 2nd form of TCR( CDS complex
together with a dimer of polypeptides γ and δ).

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• They may protect epithelial surfaces thus preventing the spread of
infection or cancer.
• These cells are either CD4- CDS- or CD4- CD8+
• These cells are predominant in certain epithelial tissues, such as the
skin. When these cells are activated in the epidermis they secrete a
cytokine that may facilitate wound healing.
• Increased number of these cells has been observed in patients with
tuberculosis.
• Macrophaqes:
• Macrophages (MQ) are released from bone marrow as immature
monocytes and mature in various tissue locations where they reside for
weeks or years. They accumulate slowly at sites of infection, respond to a
variety of stimuli (including lymphokines).
- These cells form a link between non-specific arid specific immune
response.
- They carry receptors for: Fc portion of IgG &IgA, complement
component (C3b), HLA class I and class II.
- Macrophages are activated by lipopolysaccharide (LPS) from bacteria
and IFN y secreted by activated T lymphocytes.
Functions:
a- They are active phagocytic and killer cells, working with the acquired
immune system and complement to phagocytose and kill antibody or
C3b-coated particles.
b- Macrophages play a pivotal role in the preparation of the antigen for
presentation to the lymphoid cells through digestion by its enzymes
and releasing highly immunologic parts of amtigen thus act as a major
antigen presenting cells ( APCs). They aid in the initiation and
coordination of the immune response by producing inonokines as IL-
1.
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c- They act as depot for maintenance of the immune response.
d- Important cells in delayed hypersensitivity reactions and tumor
immunity.
e- They have secretory function, they release the following major
secretory products:
• Lysozyme
• Factors that influence cell differentiation (e.g., colony-stimulating
factor)
• Cytotoxic factors (e.g., TNF- a )
• Hydrolytic enzymes (e.g., proteinases such as lipase, phosphatase,
collagenase)
• Interleukins such as IL-1 -which is an endogenous pyrogen, IL-6, and
IL-10. They also secrete chemokines. which is a large family of
cytokines that attract multiple types.of leukocytes (e.g., IL-8).
• Complement components Cl to C5, properdin, and factors B, D, I and H
• Interferon-cc
• Various plasma proteins and coagulation factors
•Oxygen metabolites (e.g., H2O2, superoxide anion)
• Arachidonic acid and metabolites (e.g., prostaglandins, leukolrienes)
• Natural Killer Cells (NK cells):
- They are large granulated lymphocytes; they are innate cytotoxic cells.
They make up 5-10% of peripheral lymphocytes.
- They are not antigen specific reactive lymphocytes; they are normally
present in the body without prior antigenic experience.
- They arise from bone marrow precursors but are of lineage distinct from
that of either T or B cells.
- NK cells receptors include Fc receptor and receptors for IL-2 & IFN y.
Functions of NK cells:

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1- NK cells are cytotoxic for tumor cells, graft cells and virally infected
cells.
2- Play a role in resistance to some bacterial, fungal and parasitic
infections.
3- NK cells are effector cells for antibody-dependent cell-mediated
cytotoxicity (ADCC) via Ab bound to their target cells through
presence of Fc receptors (CD lo) for IgG on NK cells. In addition to
NK cells, macrophages, neutrophils and eosinophils also participate
in ADCC.
4- Regulate the immune response through the secretion of numerous
cytokines.
Mode of action:
▪ NK cells kill their target by perforating the cell membrane, causing
holes or pores by performs (released from granules within the NK
cell cytoplasm) resulting in depolarization, abnormal ion flux and
essential metabolites leakage from the target cells cytoplasm. Also,
by Fas ligand-Fas pathway.
▪ They are not subject to MHC restriction (do not require recognition
of MHC molecules on the target cells), so their target range is
broad.
▪ They do not possess antigenic specificity and not acquire
immunologic memory.
▪ NK ceils secrete numerous cytokines during interaction with the
target cells including a & y IFN, IL-2 and TNF a.

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 Antigens and Immunoqenicity
Antigen and Immunogen
1- Antigen is used to describe the ability of a substance to react with the
products of the immune response (antibody or T cell). So, substances
that are recognized by a particular immunoglobulin or T-cell receptor
(TCR) can serve as the target of an immune response, and are called
antigens.
2- Immunogen is the inherent ability of a substance to induce a specific
immune response Using this definition, an antigen need not have all
the physical or chemical properties necessary to induce an immune
response. Thus immunogens are antigens, but the reverse is not
necessarily true.
*Epitopes (also called antigenic determinants) are the sites either on or
within the Ag with which Abs and T-cell receptors react. Structure: They
are very small, can be linear or conformationai. Some epitopes are
located on the antigen's surface, whereas others are internal
Function: Epitopes determine the specificity of antigen molecule.
Epitopes are able to react with their homologous antibody.
*Paratooe: Antibodies are specific for epitopes, the area of Ab molecule
that interacts with .the epitope is called the paratope. The structure of the
paratope is complementary to the epitope.
*Valence of an Antigen. Antigens are multivalent; that is, an antigen
molecule carries a number of different epitopes, some specifying Ab "A"
others Ab "B" and so forth. The valence of an antigen is equal to the total
number of epitopes the antigen possesses.
*Molecular changes: Antigen can be artificially manipulated by: altering,
adding, or taking & way epitopes. With each change, antigenicity is
altered i.e.; new antigens are produced by altering epitopes. Denaturation
or hydrolysis of the protein almost destroys conformational epitopes.

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IMMUNOGENICITY
The degree of immunogenicity of a molecule is influenced by several
factors: -
A-Foreiqnness: An antigen must be foreign to the host with which it
makes contact. The more foreign, the more to be a powerful antigen. Man
and animals do not respond to their own-self antigens because during
embryonic life the developing immunological tissues recognize self-Ags
through self-markers attached to them and develop a state of natural
immunologic tolerance to these self-Ags. However, under certain
pathological conditions an individual may respond immunologically to
Ags of his own tissues, these pathological conditions are called
autoimmune reactions or diseases.
Antigens exhibit species specificity and according to species there are
four kinds of antigens:
a- Autologous Ags, are found within the same individual (not foreign
e.g., skin graft from an individual's thigh to his chest) .
b- Syngeneic Ags, are found in genetically identical individuals (identical
twins), is not foreign
c- Allogeneic Ags, are found in genetically dissimilar members of the
same species, e.g., a kidney transplant from mother to daughter, it is
foreign. Some allo-Ags called iso-Ags, are found in some members of
a species and not in others e.g., The A & B blood group Ags.
d- Heterogeneic Ags or Xenogeneic Ags are found indifferent species
e.g., a transplant of monkey kidneys to humans, it is foreign.
B-Molecular Size:
The size of the molecule is important to its ability to induce an
immune response. Usually, the larger the molecule, the better the
immunogen, although there are exceptions. For example, insulin is small

22
but is immunogenic; carbon particles are very large but are
nonimmunogenic. Size is important for several reasons:
1-The number and variety of epitopes increase proportionately with the
size of the protein.
2-Larger molecules are easily phagocytized with the result of antigen
processing and antibodies formation. Also molecules must be
susceptible to intracellular catabolism within the phagocytic cell.
Antigens that are difficult or impossible to be phagocytized are
nonimmunogenic, e.g., polystyrene particles.
Haptens
- Structure. Haptens are unique molecules sometimes called " partial
antigens" or "incomplete antigens". They usually are too small to be
immunogenic.
- They cannot induce an immune response alone. They need to be
coupled to a large carrier molecule (e.g., protein) to be immunogenic.
However, they can react with the products of that response. Examples of
haptens include antibiotics (e.g., penicillin), analgesics (Aspirin,), and
other low-molecular-weight compounds e.g., Methyldopa; Gentamicin.
However, many complex carbohydrates arehaptenic (e.g., the capsule of
the type b strain of Haemophilus influenzae).
C-The Chemical Complexity:
It contributes significantly to its immunogenicity. The more varied the
epitope composition 1 of an antigen, the more likely different (individual)
immune response will be induced.
1-With exception of pure Hpids, most macromolecular organic chemical
groupings can be immunogenic.
a- Proteius are the strongest immunogens. Adding epitopes to the
molecule can enhance immunogenicity; lipoproteins of cell

23
 membrane induce immune response to epitopes of both lipid and
protein.
b-Polysaccharides. Most polysaccharides are nonimmunogenic.
However, certain polysaccharides can be immunogenic, e.g.
-The capsular polysaccharides e.g., pneumococcal capsule
-Lipopolysaccharides such as endotoxins that occur within the cell
membranes of Gram-negative bacteria.
c-Glycoproteins are best illustrated by A and B blood group antigens &
the Rh antigens.
These substances are strong immunogens.
d-Small polypeptides are usually weakly immunogenic, e.g., hormones
(insulin, growth hormone). Immune responses are induced only after
prolonged exposure to the antigen, or by the use of adjuvants.
e-Nucleic acids are nonimmunogenic, however, when they combine with
basic proteins they can act as immunogens. Nucleoproteins are strong
immunogens.
f- Lipids are also nonimmunogenic, although a few (e.g., lipoprotzin) can
impact specificity to antigens when presented to the immune system.
2-Antigenic mosaic. Microbial agents and mammalian cells are strong
imraunogens, and they present a vast array of different antigens to the
host. This diversity is sometimes called the antigenic mosaic of the cell.
Each antigen has multiple epitopes; therefore, many different antibodies
will be produced against cellular immunogens. Extracellular bacteria
provide an antigenic challenge to the immune system with the variety of
bacterial structures presented on their cell membrane or cell walls.
Bacteria also secrete a variety of toxins that also requirements of
immunogens. All of these bacterial components can induce antibody and
CD4 lymphocyte response.

24
Viruses also induce immune response. When viruses are extracellular,
they are capable of inducing the same types of immune responses as any
extracellular complex antigen. However, when sequestered within a cell,
the CDS component of the immune system is Iry responsible for
identification and elimination of the viral antigen.
D-Other Factors That Influence The Degree of Immunoqenicitv
1-The ability to be degraded by macrophage enzymes:
Macromolecules that cannot be degraded and processed by APCs are
poor immunogens e.g., polystyrene particles or asbestos.
2- Structural stability: Highly flexible molecules mat have no fixed
shape are poor immunogens. For example, gelatin is structurally unstable
and dose not induces an immune response unless the molecule is
stabilized by cross-linking peptide chains.
3- Dose and Route: Both the routes of exposure as well as the amount of
antigen are of critical importance in determining the type and nature of
the eventual immune response. For each antigen, there is a critical
concentration that is necessary to induce immune response. As a general
rule, the subcutaneous or intramuscular routes of antigen injection are
best for soluble substances, whereas an intravenous injection may be
more effective for cellular immunogens, such as erythrocytes or bacterial
vaccines. In practice, the intravenous route is not used because mere
exists danger of embolism and other adverse effects.
4- Adjuvants. Adjuvants are substances that enhance the
immunogenicity of molecules without altering their chemical
composition. They cause non specific stimulation of the immune
response. Adjuvants are added to vaccines to enhance immune response.
Mechanisms of action:

25
1-Increasing the efficiency of macrophage processing of antigen; by
increasing the effective size of the irnmunogen, and so promoting
phagocytosis and presentation by macrophages.
2-Prolonging the period of exposure to the antigen (act as a depot),
prolonging retention of the immunogen.
3-By stimulating the influx of macrophages or oth^ immune cell types to
the injection site.
4-By promoting local cytokine production and amplifying the
proliferation of immunologically committed cells.
Examples of adjuvants:
- Bacille Calmette-Guerin (BCG) is a live attenuated vaccine used for
prophylaxis against tuberculosis, it causes macrophage activation and
enhances NK cell activity. The adjuvant activity resides in cell wall
glycolipids (e.g., wax)
• Aluminum salts (e.g., aluminum hydroxide, and alum). An example is
alum-precipitated toxoids for vaccination against diphtheria and tetanus.
These substances retard the absorption of the antigen and thus prolong the
exposure to Ab-forming cells, they also cause local inflammation, which
increases mononuclear cell exposure.
5-Host Genetic.
The ability to respond to a particular antigen varies with genetic makeup.
For example, pure polysaccharides are immunogenic when injected into
mice or human but not when injected into guinea pigs or rabbits,

26
 Special Types of Antigens
Heterophile Antigens
-Heterophile antigens have in common one or more epitopes; this is the
basis of their antigenic relationship. As a result, they are cross-reacting
[i.e., they combine (react) with the same specific antibody because of the
presence of this shared epitope],
-The process by which cross-reacting epitopes cause immunologic
damage is referred to as antigenic mimicry (e.g., shared epitopes). An
example is tissue damage that occurs as a result of sharing epitopes
between Streptococcus pyogenes and human heart tissues in rheumatic
fever.
- Heterophile antigen stimulates the production of Heterophile-Abs.
-The principle of the cross-reacting Heterophile-Ab response has a
practical application in certain laboratory tests;-
l. Weil-Felix reaction, serum from patients with rickettsial infections
(typhus fever) agglutinates certain non-motile strains of Proteus
vulgaris (OX19, OX2, OXK).
2. Paul Bunnel test, most affected patients with infectious mononucleosis
caused by the Epstein-Barr virus, have a serum heterophile Ab that
reacts with sheep red blood cells (SRBCs)
3. Tests used for diagnosis of Syphilis. Treponemapallidum shares an
epitope with a lipid called cardiolipin (found in beef heart muscle
extract). Individuals who are infected with T.pallidum make an Ab that
reacts with cardiolipin.
Histocompatibilitv Antigens
1-In man two strong histocompatibility antigens are known which are the
ABO. and the major histocompatibility complex (MHC) antigens. These
antigens are termed human leukocyte antigen (HLA) because they are
found in high concentrations on lymphocytes and other white cells. They

27
also occur on other nucleated cells of the body (e.g., tnacrophages, and
hepatocytes)
2-The term major histocompatibility complex is a genetic term that refers
to a set of multiple genes, HLA refers to the MHC proteins in the human
and is found on the short arm of chromosome 6
3- Most of the HLA genes are polymorphic.
MHC Classes.
The HLA molecules encoded by genes of the MHC fall into 3 classes:
HLA-A, HLA-B, and HLA-C are class I antigens.
• HLA-DP, HLA-DQj and HLA-DR are class H antigens.
• Class in antigens are associated with certain complement components,
C2, C4 & factor B, also with INF a and P).
Class I antigens are expressed on all nucleated human cells and present
products of cellular synthesis to CDS cytotoxic T lymphocytes. Class II
antigens are found on a more restricted set of cells (antigen-presenting
cells, B cells, activated T cells, and, at lower concentrations, on some
other cell types) and present processed antigen to CD4 helper T
lymphocytes.
Superantiqens:
• They are called superantigens because of their ability to activate all
T cells "expressing common sequencing in their T cell receptor
irrespective of their specificity for antigen and also because they do
not need to be processed like other protein antigens. They interact
with MHC molecules outside the peptide- binding groove, and also
bind with TCR (3 chain irrespective of antigen specificity.
• Superantigens are a class of some bacterial toxins and retrovkal
proteins that have the ability to bind to both MHC class II and TCR
β chain.

28
 • Superantigens are very potent mitogen for T cells, stimulating the
release of large amount of cytokines particularly TNF.
• Examples of superantigens are some exotoxins produced by
Staphylococcus aureus, e.g., Staphylococcus enterotoxin causing
food poisoning and toxic shock syndrome toxins (TSST) causing
toxic shock syndrome. Also, pyrogenic exotoxins of streptococci.
Classes of antigens:
1- Thymus-dependent Antigens. Most humoral IRs are thymus
dependent; they need participation of Th cell to induce Ab synthesis,
e.g., erythrocytes, serum proteins, hapten-carrier complexes.
2- Thymus-independent Antigens. A few antigens are thymus
independent, they are able to induce Ab production without the aid of
T lymphocytes; Th cells. Thymus-mdependent antigens induce only
IgM Abs. There is no anamnestic response (memory) on 2ry
challenge with the same immunogens.

Humoral Immune Response

Induction of humoral immune response:
B cell activation and Ab synthesis.
• Ab production takes place in lympha^tis^^s, either in spleen or
lymph nodes or at sites of inflammation. 4 Plasma cells & B cells
are mainly responsible for Ab synthesis.
• B cell begins proliferation and differentiation into an Ab secreting
plasma cell after interaction with Ag. Secretion of large amounts of
Ab begins without change of the antigenic specificity of the
secreted Ig.
Stages of B lymphocyte stimulation include:
1. Recognition of Ag by immune cells,

29
2. Binding of Ag to Ag-R on B-cell surface in the presence of
costimulatory molecules (helper factors from an activated Th cell e.g.,
CD40 on the B-cell surface to CD40 ligand on the activated Th cell),
3. Activation of the cell through triggering of intracellular signalling
mechanisms,
4. Proliferation & differentiation into plasma cell that secretes Abs.
* Some B cells become memory cells, which are long-lived responsible
for anamnestic response (usually express IgG in their cell membranes).
* Two signals are involved in differentiation of B cells into Ab-forming
plasma cells:
1. Interaction between Ag and monomeric IgM-R on B cell surface,
2. Contact between Th cells and B cells, which is mediated by
lymphokines released from Th cells .Th cells can help only B cells
bearing identical class IIMHC (this response is MHC restricted).
• 4 Most humoral IRs are thymus-dependent (need participation of
the Th cells).
• A few Ags are thymus-independent.
• Various IL-s (B-cell growth factors) also affect B cell proliferation
and maturation.
1. IL-1, IL-2, IL-6 ———— enhance Ab production.
2. IL-4, IL-5 ———————— influence Ig class switching.
• INF y ——————— acts as stimulant to B-cell differentiation.
Adhesion molecules are important in cell activation

B-cell activation. Antigen binding to the surface imunoglobulins, coupled

with soluble or contact-(mediated nwper factors from an activated TH cell

30
Immunoglobulins
• Immunoglobulins (Ig) or antibodies (Ab) are glycoproteins present in
the gamma-globulin fraction of serum.
• Igs are produced by B cells or plasma cells in response to exposure to
Ag. They react specifically with that, Ag in vivo or in vitro; therefore, Igs
are part of the adaptive immune response (specifically, humoral
immunity).
General Functions of lgs:
1. Prevent adhesion and colonization of microorganisms.
2. Toxin neutralization.
3. Inflammatory reactions
4. Opsonization & phagocytosis of microorganisms.
5. Lysis of invading microorganisms by fixation & activation of C
components.
6. .ADCC.
7. Passive immunity to the fetus.
Ig structure:
A) Basic unit (monomer) consists of 4 polypeptide chains {2 identical
heavy (H) & 2 identical light (L) chains} linked covalenty by disulfide
bonds.
B) Heavy Oft and light (L) chains:
- H chains have a M.W. is twice that of the L chains. H chains contain
about 400 amino acids, which is twice the number in L chains.
• Isotypes (classes): amino acids differences in the carboxy-terminal
portion of the H chains identify 5 isotypes, which form the basis for the 5
classes of Ig molecules (IgG, IgM, IgA, IgE and IgD). They have the
same basic unite structure with many features in common but differ by
one or more antigenic determinants present on their heavy chains which
is specific for each class of Ig and which distinguish it immunologically

31
from other classes, These differentiating antigenic determinants on the
heavy chains have been designated by Greek letters (y) for IgG, ((a ) for
IgM, (a) for IgA, (e) for IgE and (8) for IgD.
• Subclasses: H chains classes γ&α are subdivided into subclasses (γ l, γ
2, γ 3, γ 4-αl,α2). The H chain subclasses correspond to Ig subclasses: yl
correspond to IgGl, al to IgAl and so forth.
-L chains: M.W. = 1/2 that of H chains, and are composed of about 200
amino acids.
• Types: They are of 2 types K or X based on their structural (Agic)
differences. All Ig classes have both K&A, chains. However, a given Ig
molecule may contain either 2 identical K chains or 2 identical K chains
(never a K chain or a A, chain combined).
• Subtypes: There is no isotype variation in K chains. However, there are
4 distinct A, chains (giving rise to 4 distinct subtypes).
C) Variable (V) and Constant(C) regions:
Each H & L chains has a V (shows a wide variation in ammo acids
sequence in the amino-terminal portion) & a C regions (lies in the
carboxy-terminal portion).
D) Hvpervariable regions are particular areas within the variable
regions that are highly variable in amino acids sequence. They are
important in the structure of the Ag-binding site (i.e., paratope). L chains
have 3 hypervariable regions, and H chains have 4. There are certain
areas in hypervariable regions called idiotypes, thus are part of antigen-
receptor
E) Fab (fragment Ac-binding) and Fc fragments (fragment
crvstallizabie).
The Fc portion has several properties:
• It binds the Cl components of complement and activates the
complement cascade.

32
• It reacts with Fc receptors on various cells of the body; therefore, it
dictates biologic activities of the molecule (e.g., whether a given Ig can
cross the placenta, sensitize mast cells to degranulate when they contact
allergen, or opsonize bacteria an4 other cellular elements for
phagocytosis).
-Papain split the monomeric basic unit into 3 fragments at the hinge
region (2 Fab fragments j and one Fc fragment).
-Pepsin digests away most of the Fc fragment below the interchain
disulfide bonds in the hinge region, leaving one large piece consists of 2
Fab fragments joined by disulfide bonds.
This fragment has 2 Ag-binding sites (able to bind, precipitate or
agglutinate an Ag).
F) J chains (have a joining function): are small proteins that connect 2
or more basic units in polymeric Igs (i.e., in IgM, IgA).

G) Membrane Igs: Igs exist on the membrane act as Ag receptors (IgM,

IgD). After the cell interacts with Ag, it proliferates and matures into a
cell that is actively secreting Ig. Membrane forms of IgG and IgA are
thought to function as Ag receptors on memory B cells.
H) Ig supergene or superfamilv: All members of the family share
significant amino acids sequence homology in either Ig H or L chain.
They are involved in cellular interactions. Most occur in cell membranes
and serve as recognition molecules, e.g., Ig heavy and light chains, TCR,
MHC, and many types of CD as CD2,3,4, 8, some adhesion proteins and
costimulatory surface proteins e.g., B7 and CD28.
Isotvpic variation
Igs fell into 5 classes (isotypes), based on certain structural differences.
1- Ig G:

33
It is the major Ig in the human serum (75% of total normal serum Ig pool)
at a j concentration of 1200 mg/dl.
Structure:
• IgG is a monomer (identical pairs of H & L chains linked by disulfide
bonds).
• Four subclasses have been identified (IgGl, IgG2, IgG3, IgG4) .which
correspond to H chains γl, γ2, γ3 & γ4.
• Most serum IgG is IgGl.
• Most IgG has half-life of about 21 days; IgG3 has a half-life of only 7
days,
Function:
• All IgG subclasses with exception of IgG2 can cross the placenta in
humans. Therefore, maternal IgG provides most of the protection of the
newborn during the 1st months of life.
• It is the major Ab produced in the 2ry IR.
• It has antitoxic immunity.
• It is the major opsonising Ig in phagocytosis.
• IgG, except IgG4 binds complement by the classical pathway.
2-Ig M:
It represents about 8-10% of the total serum Ig; it is present in normal
serum at a concentration of 120 mg/dl.
Structure:
IgM has a pentameric structure (5 monomeric units linked by a J chain
and by disulfide bonds in the Fc fragment).

34
Function:
• IgM is the 1st Ab that an irnrrmnologically committed B lymphocyte can
produce.
• IgM is the predominant Ab in the Iry IR to most Ags.
• It dose not cross the placenta so, an elevated IgM in the cord serum of a
newborn (Normal level =10 mg/dl) indicates fetal infection before birth.
IgM has a serum half-life of about 10 days.
• IgM is the only Ab made to certain carbohydrate Ags, such as the ABO
blood group" Ags (natural isohemagglutinins).
• Being mainly present intravascularly it protects against bacteraemia.
• IgM is the most efficient Ig at activating complement in lytic actions.
• It enhances phagocytosis by complement activation.
• Some IgM is synthesized locally in secretory tissues (e.g., parotid
glands).
3-Ig A:
It is present in 2 forms: one in the serum (serum IgA) & the other
in various body secretions (secretory IgA-sIgA). It has a serum half-life
of 6 days. IgAl_can be inactivated by an IgA protease produced by
gonococci, meningococci, pneumococci and H. influenzae. Therefore,
IgA2 is important in macoasl immunity against these pathogens.

a-Seram IgA. It is present at a concentration of 200 mg/dl, accounts for

about 15-20% of the total normal Ig pool. 80% of it exists in amonomeric
form; the predominant subclass is IgA 1. The other 20% exists as dimers,
trimers, or ietramers (linked by disulfide bonds and J chains).
b-Secretory IgA (slgA). It is the predominant Ig in various secretions
(e.g., saliva, tears, colostrum & milk and bronchial, genitourinary, and
intestinal secretions). The dominant subclass in secretion is sIgA2.

35
Structure: The slgA consists of 2 monomeric units plus a J chain and
secretory component.
* Secretory component is synthesized locally by submucosal plasma
cells as it passes to the lumen of organ in which it is produced, it acquires
secretory component. which is a polypeptide synthesized by epithelial
cells.

Function.
• Secretory component facilitates transport of slgA into secretion and
protects it from being digested by proteolytic enzymes. « Secretory IgA
acts as "gate-keeper" protects mucosal surfaces from adhesion and
colonization of microorganisms.
4-ig D:
It represents less than 1% of the total Ig poo! and is present in trace
amounts in normal serum (3-5 mg/dl).
Structure, IgD exists as a monomer.
Function. There is still controversy over the biological functions, it may
be involved as Ag receptor and in B cell activation. It has Ab activity for
a variety of haptens and Ags, such as penicillin, insulin, and diphtheria
toxoid. It has a serum half-life of 2-3 days, and is heat and acid-labile.
5-Ig E:
It is present in trace amounts in normal serum (0.05 mg/dl) s accounting
for only 0.005% of total serum Igs. Structure. IgE is a monomer.
• IgE is produced by B cells and plasma cells in the spleen, in lymphoid
tissue of the tonsils and adenoids, and in respiratory and gastrointestinal
mucosa. IgE dose not cross placenta. IgE production begins in the fetus
early in gestation.

36
• IgE has a vascular half-life of 2-3 days and is heat-labile at 56G°.
Function
• IgE is associated with atopic diseases ( immediate hypersensitivity
reactions e.g., asthma)
• IgE is homocytotropic; it has an affinity for cells (cytotropic) of the host
species that produce it (homo). This affinity is particularly strong for
tissue mast cells & blood basophils. Fixation to these cells occurs via
Fc-R
• On combining with allergens, IgE Abs triggers the release of histamine
and other mediators of atopic disease from the mast cells.
• It may also be important in immunity to certain helminthic parasites.
• IgE is unable to activate complement via the classical pathway

Phases of The Humoral Immune Response

Kinetics of The Immune Response
l-Primarv Immune Response
It occurs following the 1st exposure to an antigen (Ag), and
produces relatively small amount of Ab. It is-divided into:
A-Latent or lag period (1-2 Ws): It is the interval before Abs can be
detected in the serum. It differs according to the-type of the Ag and the
route of administration.
B-Exponential production phase: The concentration of Ab in the serum
increases but to low peak and maintained for some weeks.
C-DecIine phase: The concentration of Ab falls rapidly to the
preimmunization level.
** IgM-Ab is the 1st class of Igs that can be detected, followed by IgG-
Ab
II-Secondary Immune Response (Anamnestic response or Memory
response or Booster response):

37
- It occurs following subsequent exposure to even a small amount of
antigen.
- It consists of a rapid proliferation of plasma cells, with subsequent
production of large amounts of specific Ab.
- The Anamnestic response occurs because large populations of memory
B and T cells are recruited into the humoral IR. These memory cells are
produced during the initial exposure to the antigen. So, 2ry immune
response can be induced at any time after the Iry response, even years
later .It is characterized by: short latent period, marked increase in Ab
production with higher concentrations of Ab in the serum and gradual
decrease in Ab level.
**Ab formed are mainly IgG-class with high concentrations while IgM is
produced at the same level and for a limited period as in Iry immune
response

The IgM and IgG antibody levels in primary and secondary immune
responses to an antigen.

Monoclonal Antibodies
Monoclonal antibody is an Ab that is specific to a single epitope.
Monoclonal antibodies (MCA) are prepared by Hie murine hybridoma
technique. Hybridomas are artificially created JUWlfet produce jpure or
monoclonal atitibodies (MCA).
1-A mouse is injected with Ag, and plasma cells from its spleen are fused
with cancerous mouse cells (i.e., plasmacytoma or myeloma cells).
38
 • The cells are cultured at high dilutions to allow only one fused cell
per culture well.
• Only hybrid cell (fused) cells replicate because plasma cells are
fully differentiated cells "(which do not multiply), and the
plasmacytoma cells are susceptible to metabolic poisoning by
appropriate alterations in the media.
2-Each hybrid cell formed by this technique has 2 important properties: a-
It produces the single type of Ab molecule of its spleen cell parent
(specific for that particular epitope).
b- It continually grows and divides, like its plasmacytoma cell parent
(uncontrolled multiplication).
3-The clone of hybrid cells that produce the desired Ab is identified and
then grown as a continuous cell line, from which large amounts of the
monoclonal Ab (homogeneous) can be harvestecl.
Uses of Monoclonal Abs:.
1-Monoclonal Abs are useful in laboratory procedures,
a- For example fluorescent monoclonal Abs are used to assay specific T-
cell subsets by flow cytometric techniques.
b- A constant and uniform source of Ab with a single specificity not
only affords a powerful research tool but also can be expected to
provide quicker and more accurate method for identification of
viruses, bacteria, and cancer cells.
2- Monoclonal Abs have been also shown to be therapeutically useful in
the treatment of cancer, MCA are produced against tumor-specific
antigen and are used either alone or after binding with anticancer
agents such as radioisotopes or cytotoxic agents.
3-Monoclonal Abs may represent an alternative source of immunogens
for human antadiotypic Ab vaccines.

39
4-Preparation of antisera against antigenic determinants in bacterial, viral
or tumor antigens
5-Passive immunization and immunoprophylaxis against HBV, clostridial
toxins and others.
6-Preparation of monoclonal Abs against HLA-Ags to be used in HLA
typing.
7-Monoclonal Abs are used as a source of IgG anti-RH (D) to prevent
hemolytic disease of newborn.
8-Monoclonal Abs are used in isolation and purification of human
products prepared in bacteria or yeast by genetic engineering e.g.,
vaccines, IL-s and IFNs.
9-In assay of hormones.
10-Administration of anti-CD3 Ab in cases of kidney transplant patients
to inhibit T-cells causing acute rejection.
Production of monoclonal anibodies

The Complement System

• The complement system plays a major role in host defense and the
inflammatory process (through innate and acquired immunity). (Jt,
consists of complex series of at least 25 plasma proteins (9 major
proteinssCl-C9) are normally functionally inactive.
• It is activated sequentially in a cascading manner, each protein
activating the protein: directly follows it in sequence.
• It causes lysis of erythrocytes in hemolytic anemia, sensitizes
foreign particles to phagocytosis, and causes release of histamine
from mast cells.
• It is present in serum and tissue fluids except urine and CSF.

40
 • It is present in high concentration in guinea pig sera.

Metabolism
l-Synthesis. The liver is the major site of synthesis of C proteins,
macrophages and fibroblasts .can also synthesis some proteins as well.
Complement proteins 1st appear in the fetus during the 2 month of
pregnancy. At birth the concentration of most components is about 50%
of adult levels. Inflammation increases the synthesis of complement
components, through the suction of IL-1 and IFN y. 2-Catabolism. The
catabolic rate of C proteins is high, 1%- 3% per hour.
1- Synthesis. Genes for C proteins (factor B, C2, C4) are located in
MHC on human chromosome number 6 in class III. Genes of six of
the C receptors and genes for the •regulatory proteins are located
on chromosome 1.
Pathways of complement activation
Complement activation occurs in 2 pathways:
1-Classic pathway:
It is evolved mechanism of specific adaptive immunity.
• It is initiated by Ab, usually bound to Ag(Ab coated bacterial cell, a
tumor cell or a lymphocyte) or by aggregated Igs (IgGl, IgG3 and IgM),
• It requires the interaction of all nine C components. Activation follows
the binding of Cl to Fc fragment of Ig (CH2 of Ig G in the presence of
Ca++).
• As a result of fixation of complement it becomes activated with the
following sequence Cl, C4, C2, C3, the last split into C3a and C3b then
followed by C5-C9 with the final stage is cell membrane attack with lysis
and death of the cell, when C5b6789 complex forms.
• Activation of Cl- C5 occurs through enzyme cleavage, while activation
of C6-C9 occurs through covalent binding.
41
Pathways of complement activation

2- Alternative pathway (Ab-independent):

• It provides nonspecific innate immunity.
• It can be initiated by lipopolysaccharides or endotoxin from cell
walls of gram negative, bacterial, by tumor cells or by aggregated
IgA or IgG4 cell. It dose not require components Cl, C4, and C2.
• With certain plasma factors e.g., factor B, factor D, and properdin
(factor P) enhancement of cleavage of C3 into C3a and C3b, then
the sequence of activation will be the same as in the classical
pathway ending in killing and lysis of the target cells.
Regulatory mechanisms:
Complement activation is associated with potent biological
functions that if left unchecked, exhaust the complement system and can
cause significant damage to the host. Several types of protein-
complement interactions prevent uncontrolled activation of the
complement system.
A-Inactivators: e.g., Factor 1 (C3b inactivator), and anaphylatoxin
inactivator (inactivates C3a, C4a and C5a).
B-Inhibitors: e.g., Cl esterase inhibitor, and factor H (act with factor I to
bind C3b).
C-Regulatory proteins in the cell membranes; Inactivate membrane-
bound C3b, C4b or C5b67.
Biological activities of complement components
1-Chemotaxis: C5a. attracts phagocytic cells to the site of reaction and
increases their activity.
42
2-Anaphvlatoxin: C3a & C5a, stimulate the release of mediators of
inflammation from mast cells and basophils leading to dilatation of
blood vessels and increased capillary permeability with influx of
more Abs and C to control the inflammatory reaction.
3-Opsonization: C3b coats the participate Ag, render it more attractive to
phagocytes that have receptor for C3b.
4-Cytotoxic or cytolytic effect: C8 & C9 lyse the cell by nonenzymatic
process similar to the membrane damage caused by the perform
secreted by NK cells and Tc cells
Complement however may be responsible for damage of the body's
own cells in immune complex diseases (e.g., in type III hypersensitivity
reactions and also in type II hypersensitivity reactions).
Practical applications. An intact C system is essential to maintenance of
health.
1- Estimation of C components is useful in clinical practice as indicators
of disease activity. It is important for diagnosis, management of
patients who present with recurrent infection, allergic diseases, and
autoimmune diseases.
2- Decrease in C serum level may be acquired or inherited leading to
recurrent infections.

43
 Cell mediated immunity
There are two forms of cell mediatd immunity (CMI), non-specific
and specific. Non-specific CMI occurs by NK cells,, macrophages and
granulocytes. These cells respond to a variety of stimuli in a non-specific
manner. In contrast, specific CMI is conducted by T. In this condition
antigenic stimulation select and activates particular cells that possess
recptors with specificity to that antigen. So only particular cells become
activated, these cells proliferate and produce specific clones of effector
cells and memory T cells with identical i antigen recognition specificity,
Several cell generations and several days must elapse before expression
of CMI reactions. A secondary CMI displays similar characteristics to a
secondary humoral immune response in that it is usually more
pronounced and occurs more rapidly.
CMI unlike Humoral Immunity (HI):
It is not transferred to a non immune individual by serum, but
delayed hypersensitivity can be transferred from sensitized to non
sensitized individual with Ag reactive cells
•CMI is delayed in time (24 - 48 hours), while HI is seen in minutes after
the secondary exposure to the same Ag.
•CMI is initiated by intracellular Ags.
•CMI reactions are characterized by significant mononuclear cell
infiltration with resultant duration. While, Ab-mediated reactions have
more fluid and erythema (wheal and flare).
MI is initiated by Ags, which are held locally in tissues. Thus CMI occurs
in: Infections with intracellular pathogens i.e., microorganisms capable of
living and multiplying within the cells and the host. For example Sonie
bacterial infections as Mycobacterium tuberculosis, Mycobacterium
leprae, Brucella, Lymphogranuloma venereum.and Rickettsiae

44
Viral and fungal infections.
1. Parasitic infection as toxoplasmosis arid leishmania.
2. Delayed hypersensitivity reactions.
3. Contact dermatitis.
4. Allograft rejection.
5. Some autoimmune diseases.
6. Resistance to tumors.
CMI has 2-Effector T cells (TH1 cells and Tc ceils).
1-Thl (delayed hypersensitivity cells = TD) recognize intracellular Ags,
secrete γ IFN and other cytokines, activating macrophages to cytotoxic
activity & enhancing other T and B cell responses. DH cells are
CD4+cells.
2-TC cells recognize foreign Ags on most cells of the body, secrete y IFN
which induces other cells to produce antiviral proteins. They also have
direct destruction of target cells (graft tissue, tumor, virus-infected cells)
by releasing perform with subsequent cytolysis (direct cytolysis). Tc cells
are CD8+cells.
Mechanisms of CMI Response
Antiqenic processing and presentation
• Uptake, processing, presenting of Ag and activation of T cells.
• Ag-processmg cells and T cells can interact only if the 2 participants are
MHC identical e.g., CD4+ cells recognize Ag in conjunction with class II
MHC molecules & CD8+ cells recognize Ag in conjunction with class I
MHC molecules.
• Th cells release lymphokines (ILs) which aid in regulation of
humoral as well as CMI responses,
T-cell Activation:
1- Activation of Thl by Ag requires 2 signals mediated by macrophages:
a) Presentation of Ag in conjunction with class II MHC molecules.
45
b) Synthesis and release of IL-1. IL-1 induces the synthesis of IL-2 by T
cells and other ILs leading to full T cell activation. The responding T
cells synthesize more IL-2 Rs (CD25) forming autocatalytic cycle.

* Full T-lymphocyte activation requires two signals:

a-T-cell-receptor / MHC (signal 1) and
b-The costimulus; B7 on the APC & CD28 molecule on T lymphocytes
(signal 2).
In addition, T cells express a number of adhesion molecules (ADM) such
as intracellular adhesion molecules (ICAM-1) and leukocyte function-
associated Ag (LFA-1) that bind Hgands on the surface of APC and thus
promote cell-cell contact.
• Stimulation of Thl cells by intracellular microorganisms leads to type
IV hypersensitivity,

Th cell activation
2-Activation of Tc cells by Ag in association with class I MHC
molecules. Two signals are required:
a) Interaction between TCR-CD8 complex on TC cell and specific
epitope class I on APCs.
b) Release of ILs by macrophages and Th cells.
Thus Tc cell activation has 2 levels of MHC restriction:
i) Class I restriction in epitope presentation to TC cell,
ii) ii) Class II restriction in epitope presentation to Th cell.

46
Tc cell activation
3-Activation of Ts cells, Ag presentation by macrophage may not be
required, Ag is capable of activating Ts cells directly. Ts activation does
not appear to be under MHC control.
Consequences of CMI:
-Cell mediated immune response to antigenic stimuli is associated with
both immunity and inflammation.
-CMI has been termed delayed hypersensitivity or cell mediated
hypersensitivity.
-Consequences of CMI are exemplified by Koch's phenomenon. Koch
observed that guinea pigs infected with tubercle bacilli showed after 2
weeks at the site of injection (subcutaneous) a firm nodule, which
ulcerates and the ulcer dose not heal. This is accompanied by
enlargement and caseation of the regional lymph nodes.
-Reinfection at a different site by another dose of tubercle bacilli some
weeks later, results in formation of a nodule at the inoculated area
within 2 days, which soon ulcerates but the ulcer heals quickly while
the local lymph nodes are unaffected.
-Thus in reinfection two phenomena are observed:
1-The development of a rapid reaction at the site of injection i.e.,
hypersensitivity.
2-Infection remains localized without spread to lympji nodes, an4,quick
healing of the ulcer i.e., immunity.
-The classical example of CM response in humans is tuberculin skin
reaction., A positive reaction parallels the hypersensitivity in Koch's
phenomenon in guinea pigs.

47
CYTOKINES
• Cytokines are intercellular regulatory proteins that mediate a
multiplicity of immunologic as well as non-immunologic functions.
Cytokines regulate not only local and systemic immune and inflammatory
responses but also wound healing, hematopoiesis, and many other
biologic processes
• These proteins are induced by a variety of different tissues and
individual cells and arc induced by specific stimuli to modify and
enhance the function of effector cells and are regulated by feedback
inhibition or by production of other cytokines.
• Cytokines produced by lymphocytes are called lymphokines whereas
those produced by monocytes and macrophages are called monokines.
• Most cytokines have more than one biological activity. A single
cytokine is often produced by more than one cell type and has diverse
effects on different target cells i.e., pleiotropism.
• Cytokines can be used as a marker of various diseases.
• Administration of cytokines or their inhibitors is a potent approach for
modifying biologic response associated with disease.
Cvtokines are involved in:
1-Activation of cells of the immune system: Since the function of the
immune system is the; elimination of antigens, as the majority are in
resting phase, lymphocytes must be activated and induced to proliferate
by antigens and cytokines before they can participate in effector phases of
the immune response.

48
• As well as requiring presented antigens; T cells need cytokines to
induce division and differentiation into activated cells e.g., IL-1. IL-2, IL-
4, IL-6 and IFN γ. B cells also require antigen for activation, although
they may respond to T independent antigens without further signals;
response to T dependent antigens need additional signals, including IL-4,
IL-5, IL-6 and IL-1.

Cytokines & B cell Activation

For example: macronhages presenting antigen to T cells (Th cells) and

secrete IL-l, which stimulates Th cells to secrete IL-2 and enhance the
expression of IL-2 receptor. ^2 also stimulates the growth of activated T
cells (to undergo blast transformation, division and differentiation). It
also stimulates Tc cells. It can act as a growth and differentiation
fadSiiJfer B cells and NK cells (enhances their cytolytic function
producing lymphokine-activated killer cells) and can also activate
macrophages.
2-Role in inflammation:
Infection, tissue injury and malignancy trigger an acute phase
response. It is characterized by fever, increased vascular permeability and
an increase in the synthesis of acute phase proteins (C-reactive protein,
al- glycoproteins, al-antitrypsin, o2- macroglobulin and serum amyloid A
& P, complement factors C3, C9 and B) by hepatocytes when exposed to
certain cytokine as IL-1, 6 and TNF. IL-1 stimulates hypothalamus
leading to pyrexia, thus IL-1 is called endogenous pyrogen.

49
3-Promotion of hematopoiesis:
Cytokines that stimulate expansion & differentiation of bone marrow
progenitor cells are called colony-stimulating factors (CSFs) e.g.,
• IL-3 stimulates the growth of pluripotent stem cells,
• IL-7 acts on stem cells progenitors committed to the B lymphocyte
lineage,
• IL-5 stimulates eosinophils growth & differentiation.
Also granulocytes & monocytes colony stimulating factors (GM-CSF)
stimulate the growth of haemopoietic cells that are committed to granular
or monocytes lineage, platelets & progenitors of RBCs.
4- Cytostatic and Antiviral Activities:
Cytokines involved in inhibition of cellular proliferation e.g., tumor cells,
are lymphotoxin (cytotoxic or cytostatic). They have the ability to lyse
certain tumor cells, are known as TNF α & β.
Some Important Cvtokines:
• TNF- α (Cachectin): The principal cell sources are activated
macrophages, T cells , NK cells and other somatic cells.
-It leads to vasodilatalion, edema and activation of neutrophils. It has
antitumor effect.
-It has IL-1-like effects and considered as mediator of wasting syndrome
"cachexia", which often accompanies chronic parasitic and viral infection
or malignancy. Cachectin inhibits lipoprotein lipase, preventing the
uptake of triglycerides.
-It induces other cells to release IL-1. IL-6 & IL-8 and IFN β.
-Its level increases with fever, cachexia, and disseminated intravascular
coaglulation (DIG).
" TNFβ (lymphotoxin) : Produced by activated Thl cells.
-It has IL-1-like and TNF-like effects.

50
* IFN α and β : Produced by macrophages, neutrophils and other somatic
cells.
-They have antiviral effects.
-Induce class IMHC on all somatic cells.
-Activate macrophages, NK cells and CD8+ cells.
IFNγ : Produced by activated Thl and NK cells. It's major effects are:
1-Inhibition of viral replication (antiviral effects).
2-Induction of class IMHC molecules on all somatic cells & class n MHC
molecules on APCs and some somatic cells.
3-Increasing the expression of Fc receptor on macrophages.
4-Activation of macrophages, neutrophils and NK cells.
5-Regulation of cell-mediated immunity (e.g., it inhibits Th2 cells). -
Influencing class switching in Ig synthesis.
Example of Some Important Interleukines
IL-1:
- It is a soluble factor produced by monocytes, macrophages and other
APCs. It has proinflammatory and immunomodulatory activities. When
locally produced at low concentrations, the predominant effects of IL-1
are probably immunoregulatory,
- It enhances the proliferation of CEH-4 T cells (costimulation of APCs
and T cells) and the growth and differentiation of B cells. It induces the
synthesis of IL-6.
-Systematic IL-1 shares with TNF the ability to cause fever, to induce
synthesis of acute phase plasma proteins by the liver and initiate
metabolic wasting (Cachexia).
IL-2:
- The major cell type that produces IL-2 is the activated Thl cells, Tc cells
and NK cells.
- IL-2 is the major autocrine growth factor for T lymphocytes.

51
- IL-2 stimulates the growth of NK cells and enhances their cytolytic
function, producing the so-called lymphokine-activated killer (LAK)
cells.
- IL-2 acts on B cells both as a growth factor and as a stimulus for
antibody synthesis.
- The principle clinical use of IL-2 is for cancer immunotherapy.
IL-4:
-IL-4 is a growth and differentiation factor for B cells. It is the main
cytokine capable of inducing switching of B cells to the Ig E isotype.
-IL-4 is an autocrine growth factor for a subset of cloned CD+4(Th2) that
secrete IL-4, IL-5, and IL-6.
-IL-4 is a growth factor for mast cells and synergies with IL-3 hi
stimulating mast cell proliferation.
- It inhibits Thl cell differentiation.
- Inhibition of monokine production by macrophages.
IL - 6
It is synthesized by activated Th2, APCs and other somatic cells (e.g.,
vascular endothelial cells, fibroblasts and other cells), it induces fever and
acute-phase response of liver,
- Growth stimulation of activated B-cells and Ig production.
IL-8
- It is produced by macrophages, skin dermal fibroblasts, endothelial cells
and monocyte.
- It is a member of the chemokine family of cytokines that attract multiple
types of leukocytes.
- IL-8 is a potent chemotactic factor for neutrophils, basophils and T
cells.
1L-10:

52
- It is mainly produced by activated Th2, CDS T, B lymphocytes and
macrophages.
- It inhibits T cell-dependent immune responses, acting on macrophages
(APCs)
- It inhibits cytokine production by Thl cells, NK cells, and APCs.
- IL-10 is also a growth factor for B lymphocytes.

Types of Immunity and Immunoprophvlaxis

Immunity may be acquired by natural or artificial processes
A-Examples of naturally acquired immunity includes the immunity
that develops during the placental passage of Ab from mother to fetus
(passive) and during convalescence from an infection (active).
B-Examples of artificially acquired immunity includes vaccination
(active) or injection of gamma globulin (passive).
Immunization describes the process of administrating antigen or
antibody to a live host with the purpose of inducing protection. The goal
of immunization in any one individual is the prevention of disease caused
by infectious agents. The goal of immunization of population groups is
the eradication of disease.
Vaccination is the term used for the second reason (public health), but the
terms are sometimes used interchangeably. Other applications for
vaccination:
1. Protection and treatment of allergy.
2. The prevention/treatment of tumors, e.g., tumor specific Ag vaccine.
3. Therapy of some autoimmune diseases as multiple sclerosis.
Types of Immunization
Agents used for immunization can be divided into two categories
based on the type of immunity that they induce.
1-Active immunization (immunoprophylaxis) uses vaccines.

53
 • Vaccines are able to induce active immunity and immunologic
memory without producing disease
• Vaccines consist of either intact microorganism (e.g., killed or
attenuated), or their immunologic components (e.g., polysaccharide
capsule) or products (e.g., bacterial toxoids).
• The recipient actively participates and makes Abs or specifically
reactive T lymphocytes in response to the vaccination.
• In general, attenuated vaccines will induce both humoral and CMI,
whereas killed isms or their products usually induce only Abs.
2- Passive immunization involves serum preparations.
• Passive immunization provides protective Abs for a particular
disease, it does not stimulate an immune response. Thus, it is
effective and acts immediately, but is temporary in its effect.
• It involves three basic types of serum preparations: .
1. Antitoxin (antisera)
2. Immune globulin (gamma globulin) ;,
3. Specific immune globulin
4. Monoclonal antibodies
Routes of administration
1- Injection is the most common method,
a- Intramuscular and subcutaneous injections are used most often,
b- Intradermal (intracutaneous) injection.
2- Oral is used in (oral polio vaccine e.g., Sabin vaccine).
3-Intranasal administration (e.g., influenza vaccine), this method
stimulates the immune response that mimics the response induced by
natural infection but, the protection induced is not long-lived.
Active Immunization
Bacterial vaccines

54
1- Intact bacteria may be either dead (e.g., killed) or alive (e.g.,
attenuated).
(a) Killed bacterial vaccines are produced using cultured organisms that
are killed with heat (55-60°C) or chemicals (acetone, formalin, phenol
and merthiolate). Examples of killed vaccines: TAB vaccine against
enteric fever (heat)
• Salk vaccine against poliomyelitis (formalin)
• Semple's vaccine against rabies (phenol)
• Pertussis vaccine against whooping cough (merthiolate)
Advantage:
- Cheap, easy to produce, does not need cool storage.
- Does not revert to virulence.
Disadvantage:
- Incomplete protection as a result of destruction of some Ags during
killing.
- Toxic effect due to release of bacterial endotoxins during its
preparation.
- It may cause fever and local reaction.
(b) Live attenuated bacterial vaccines are prepared by frequent
subculture in the laboratory under unsuitable environmental conditions or
using artificial unsuitable media. They provide a superior and longer-
lived immunity, however they do have some risks (revert to a virulent
state, fatal infections). Examples:
• BCG vaccine against T.B.
• 17D vaccine against yellow fever.
• Measles
• Polio (Sabin) Advantage:
- It can be given orally or intranasally.
- It causes local and systemic immunity.

55
- It provides longer and higher protection than killed vaccine
Disadvantage:
- It is expensive, difficult in preparation, need storage in cold conditions.
- It may revert to virulence.
- It may cause fever and local reaction.

Comparison of Live with Killed Polio Vaccines

Characteristics Live (OPV) Killed (IPV)
Induces Ab immunity Yes Yes
Serum IgM & IgG Yes Yes
SIgA Yes No.
Induces CMI Yes No.
Induces gut immunity Yes No.
Potential to revert to virulence Yes No.
Spread through the community Probable No.
Stability Low High
Route of administration Oral l.m

OPV= Oral polio vaccine (Sabin vaccine), IPV= Inactivated polio vaccine (Salk vaccine)
II-Microbial products, are either structural components or toxoid.
(a) Structural components (subunit vaccines) prepared by selection of
most immunogenic part of bacteria.
1. The capsule of certain bacteria (e.g., Streptococcus pneumoniae &
Haemophilus influenzae type b; Polysaccharide vaccines) has proved to
be valuable in vaccination. It induces the production of anti capsular Abs
that neutralizes the antiphagocytic effect of the capsule. .
2. Adherence pili.
3. The first vaccine to prevent hepatitis B virus (HB) infection was
composed of the surface Ag (HBsAg) isolated from plasma and

56
disinfected. Antibodies reacting with this Ag (HBsAg) and neutralizing
the infectivity of the virus. Now recombinant DNA vaccine is available.
(b) Toxoids are non-toxic, immunogenic preparations produced from
toxins and induced antitoxic Abs. Heat or formalin can be used in their
preparation. Examples: C.tetani & C. diphtheria. Diphtheria toxoid is
commonly combined with tetanus toxoid and pertussis vaccine as DTP.
The immunization with toxoid is effective for one year or more. Booster
immunizations enhance the effectiveness of the original immunization.
Adjuvants are sometimes used to prolong the antigenic stimulus and
enhance its immunogenicity. They are either incorporated with or injected
simultaneously with vaccines. They are also used with killed, subunit,
synthetic, and antiidiotypic vaccines. Examples:
1-Alum- precipitated toxoid (toxoid precipitated by K aluminum sulfate,
washed, suspended in saline), it stimulates Th2 cells.
2- Adsorbed toxoid (toxoid adsorbed onto aluminum OH or aluminum
phosphate).
III-Conjugate vaccines (polysaccharide/protein carrier).
They provide immunity against e.g., meningitis caused by H. influenza
type b in young children. The carrier protein used is tetanus toxoid, which
enhances immunogenicity of the capsular material. Conjugate vaccines
are also used to provide immunity against other encapsulated bacteria'
(e.g., Pneumococci. Meningococci, and Klebsiella pneumoniae).
IV-Svnthetic vaccine, peptide chain similar in structure to a protective
antigen of the organism is prepared as in Haemophilus influenza type b
and meningococcal vaccines.
• They lack conformational adequacy for immune response stimulation.
• T and B cell epitopes should be present.
A cocktail of peptides should be included in a general vaccine to cover
variations in individuals.

57
V-Recombinant vaccines:
They are prepared by selecting the gene responsible for the production of
the most effective immunogen and inserted into plasmids of either
bacteria as E. coli or yeast. The effective immunogen is harvested and
purified and used as vaccine (e.g., hepatitis B vaccine, meningococcus
and vibrio cholera vaccines).
VI- Anti-idiotypic Ab vaccine:
• Ig idiotypes representamino acids sequences in the hyper V region
associated with Ag-binding capacity of the molecule. Idiotypic sites are
capable of inducing Ab production (i.e., anti-idiotypic Abs).
• Anti-idiotypic Abs are able to function as Ag in vaccines against
infectious diseases, because this preparation contains an internal image
component that mimics the epitope of the microbial Ag.
Advantage: safer, substituted polysaccharide antigens (i.e., provide an
alternative approach when the antigen is difficult to isolate or
unavailable).
Disadvantage: non-immunogenic in infants, and poly-immunogenic in
adults.
• Examples: rabies, polio, herpes and hepatitis B.

Passive immunization
Passive immunity is effective and acts immediately .however, its effect is
temporary. The individual does not make Ms Abs or sensitized
lymphocytes but receive it already prepared.
Types of passive immunization:
A-Natural Passive Acquired Immunity:
It results from passage of Igs from mother to fetus through the
placenta (IgG), in colostrum (IgA, IgG) or breast milk (IgG & IgA).
B-Artificial Passive Acquired immunity:
58
• It provides rapid protection but of short period,
• It is used in medical practice either for treatment or for prophylaxis.
Three basic types of serum preparations are used:
I)-Antitoxins (antisera) consist of toxin neutralizing Abs that are
specific for a given toxin. They have no effect on microorganism that
produced the toxin. Most commercial preparations are produced by
immunizing human volunteers, horses, or cows with toxin or toxoid. The
serum or plasma is harvested and processed to concentrate the antitoxin
and eliminate bovine serum protein. Then sterilized by filtration, treated
with a chemical preservative and standardized. Such animal-derived
antitoxins are used only when a specific human immune globulin is not
available because, being foreign proteins they can cause adverse effects
such as ahaphylactic shock, serum sickness or disseminated intravascular
coagulation (DIG) and transmission of blood borne infections.
Representative currently used preparations
1 -Botulism antitoxin is prepared in horses or in human volunteers and is
administered as treatment for botulism. It contains polyvalent antitoxin to
three types of toxin (types A, B, and E) produced by Clostridium
botulinum.
2-Diphtheria antitoxin is prepared in horses by injection of toxoid of C.
diphtheria. It is used in treatment of diphtheria.
3-Tetanus antitoxin consists of human-derived immune globulin that is
specific for the tetanospasmin toxin of C. tetani. Although animal-derived
antitoxins are available, they are not recommended because of possible
adverse effects.
4-Antivenins for snake bites are prepared by injecting horses with
minute doses of the toxins harvested from the snakes.
II)-Immune globulin (gamma globulin

59
It is prepared from pooled, healthy adult human plasma or serum by cold
ethanol fractionation. It contains Abs (mainly IgG) to several
microorganisms and is used for passive immunization against various
diseases or for maintenance of immunodeficient individuals. Human-
derived gamma globulin may be used in the treatment of hepatitis A and
measles.
III)-Specific immune globulin
It is a gamma globulin obtained from people who have recently recovered
from a specific infectious disease, or from hyper immunized human
volunteers. Representative preparations include the following:
1- Hepatitis B immune globulin (Ig).
2- Rabies Ig.
3- Tetanus Ig & human-derived botulism Ig.
4- Varicella -zoster Ig,
5- Rho Ig (Anti-D serum)
VI)-Recentlv monoclonal Abs are used e.g., immunotoxins can be
produced by linking toxins to highly specific monoclonal Abs. Toxin
(e.g., ricin which is extracted from castor-oil plants) is used in cancer
therapy coupled to monoclonal Ab specific for tumor cell Ags to inhibit
protein synthesis causing cell death. Another potential use for
immunotoxins is in the management of autoimmune disease.

Adverse reactions to immunoprophvlaxis

A-Adverse effects specific to live vaccines
1-'Revert" to a virulent state, and may cause disease. It should not be
given to pregnant women (because of possible fetal infection) unless the
woman is planning to travel in an area endemic for a particularly serious
disease.

60
2-It can cause serious and even fetal infections in immunologically
incompetent individuals with the following conditions:
a) Congenital defects in CMI (e.g., Di George, syndrome).
b) Acquired defects in CMI (e.g., leukemia, lymphoma or AIDS).
c) Bruton's agammaglobulinemia
d) Treatment with immunosuppressive agents (e.g., corticosteroids,
alkylating agents, radiation or other immunosuppressants).
B-Other adverse effects
1-Allergic reactions to vaccine components such as egg proteins (e.g., in
measles, mumps) or to antibiotics (e.g., viral vaccines) may occur.
2-Serum sickness may result from injection of large amounts of foreign
proteins such as diphtheria antitoxin.
3-Transmission of blood borne infections
4-Sever reactions, such as, high temperature, convulsions or coma may
occur.

Immune response to infection

There are 2 factors that influence the immunity to infections:
1-The pathogen with its ability to enter and survive in the host.
2-The immune system of the host, which struggles to limit the infection
or its harmful products. The outcome of the contact of any infectious
agent with the host may be either:
• Infection without disease, this means subclinical infections or
• Infection with disease, this includes clinical effects, due to the pathogen
or its toxic product or effects may result mainly from hypersensitivity of
the host to the pathogen and its products.

Immune response to bacterial infection

61
Immune response to bacterial infection includes both non-specific and
specific immunity.
Non specific (innate) immunity is sufficient to resist infection by
organisms of low virulence. It includes anatomical and physiological
barriers, biologically active substances and phagocytosis-as-discussed-
before.
Specific immunity includes both humoral and CMI.
The cells and molecules of the immune system work together as an
integrated system to eliminate the infectious agent and provide immunity
that may be long lasting protective immunity.
A- Protective humoral immunity:
The presence arid persistence of antibodies in the serum, through the
presence of memory cells, during disease help in recovery. Abs can act to
prevent infection by: 1) Prevention of attachment of bacteria to the
mucous membranes. The ability to adhere to the mucosal surface plays an
important part in determining the pathogenicity of some bacteria e.g.
enteropathogenic E.coli. Specific slgA Ab is responsible for local
immunity (by coating the external surface of bacteria and viruses)
preventing their attachment to the mucous membranes.
2) Neutralization of toxins. Abs can neutralize exotoxins produced by
some bacteria (e.g., C.diphtheria, Cl.tetaoi and Cl.botulinum). Antitoxin
property is predominantly associated with IgG. Bacterial exotoxins bind
firmly to their target tissue thus they cannot be neutralized by subsequent
administration of antitoxin. Hence, in diseases mediated by exotoxins as
diphtheria, tetanus, gas gangrene, and botulism immediate administration
of antitoxin is necessary to prevent attachment of additional exotoxin and
thus prevent damage caused by the exotoxin. An important example is the
presence of antitoxin Abs against tetanus in pregnant mothers that
adequately immunized resulting in transmission of this antitoxin to her

62
fetus thus affords protection against development of tetanus neonatorum
particularly in conditions with unclean obstetric environments.
3)Immune phagocytosis and opsonization:
4)-Lysis via complement-activation (through classical pathway)
5)Antibody dependent cell-mediated cytotoxicity (ADCC):

B- Protective cellular immunity:

Bacterial infections caused by obligate intracellular pathogens e.g.,
Mycobacterium tuberculosis & Mycobacterium leprae, Brucella,
Chlamydia, and Rickettsiae are generally controlled by CMI, as antibody
can not reach the organism present in the host cells. It involves:
1- T lymphocyte killing. As a result of contact between the sensitized T
lymphocytes and an Ag, stimulation of T cells occurs with the production
of lymphokines and generation of cytotoxic (Killer T lymphocytes).
These cells confer resistance by direct destruction of target cells by
release of performs, with subsequent cytolysis.
2 - Macrophage mediated cytotoxicity: Intracellular microorganisms can
survive and proliferate in macrophages. When macrophages are activated,
these organisms are killed, as there is an enhanced intracellular killing
activity of macrophages. Macrophages are activated by T cell
lymphokines e.g. macrophage activating factor (MAF), This response is
nonspecific, so they are capable of destroying a wide range of
microorganisms.

63
Summary diagram. Mechanisms of target cell destruction.

Immune response to viral infections

Immune response to viral infections includes both non-specific and
specific immunity.
A- Non-specific defense mechanisms: (innate or natural): Just as non
specific mechanisms to bacterial infections besides:
Interference: It is the inhibition of viral replication by the presence of
other viruses, it is mediated by interferon production.
Interferons (IFNs).
• IFNs are glycoproteinss which inhibit cellular metabolic activity that is
important for viral replication. Thus they stop the viral infection at the
intracelular levels.
• IFN is species specific but not virus specific.
• IFN is released from virus infected cells within few hours after viral
invasion.
• INF is important in the early stages of infection before the initiation of
the specific immune response.
• IFN is of significant importance in superficial viral respiratory
infections in which recovery may take few days before any Ab is
detected in the blood or in respiratory mucosa e.g., common cold,
influenza.
B- Specific defense mechanisms:
1 - Humoral antiviral immunity: Viral Abs can prevent infection by:
a- Prevention of attachment and penetration; Virus neutralizing
antibodies react with viral antigens preventing the attachment
(adsorption), penetration and invasion of the cell by virus.

64
 • Humoral antiviral immunity is effective only against extracellular
viruses. It is effective in cases of infections associated with
viraemia e.g., poliomyelitis, and measles.
• In superficial infections e.g., common cold, influenza, the locally
produced slgA is important to prevent infection of mucous
membrane, which is preceded by interferon production that acts as
the earliest defense mechanism.
b - Opsonization.
c-Complement mediated lysis: Antibodies and complement can destroy
virus-infected cells.
They are active against free virions and against viral antigens expressed
on the surface of infected cells.
d-ADCC: It is an important mechanism of destruction of virus infected
cells.
3- Cell-mediated antiviral immunity:
Although serum Abs and complement are capable of neutralizing free
virus and destroy virus-infected cells, CMI mechanisms are the most
important in controlling viral infections. Recovery from systemic viral
infections such as measles, chickenpox, smallpox and herpes is due to
CMI in conjunction with immune interferon.
Cell mediated antiviral immunity acts by three mechanisms:
1 - Cytotoxic T cells; which are the major contributors to destruction of
virus-infected cells through direct cytotoxic reaction. Virus-infected cells
develop new antigens on their surface membranes. These new antigens
may arise as a result of viruses budding from the cell surface or as a result
of the cell synthesizing virus-coded proteins or modification of some
surface antigens by the virus or as a result of the cell synthesizing virus-
coded proteins or modification of some surface antigens by the virus.

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2 -NK cell activation via interferon IFN enhances NK cell activity
(spontaneous cell mediated cytotoxicity) and increases the expression of
Fc receptors on NK cells and mA-antigens class I on virus infected cells,
potentiating the cytotoxicity mediated by NK cells. This is an important
mechanism by which interferon interferes with the viral growth prior to
the development of specific cell mediated cytotoxicity.
3 - Macrophage activation (by tympkekines): Activated macropfaages
also exert a significant antiviral activity. Macrophages are activated by
IFN produced both by vires- infected cells and by T cells interacting with
the virus, which also produce MAP.

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 Contents
Subject Page
Immunity and the Immune System
Immunity (Resistance)
Tissues and cells involved in immune response
Antigens
Humoral immune response
Immunoglobulins
Phases of humoral immune responses
Monoclonal antibodies
Complement
Cell mediated immunity
Cytokines
Immunoprophylaxis
Immune response to infection

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