MEDICAL

MICROBIOLOGY I

DR. KELI, MBChB.

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 Immunology I
SCOPE ; definitions, basic concepts of immunology, overview of the
immune system ,innate and acquired defense mechanisms, antibody
and T cell receptor structures, complement system, antigen recognition,
vaccination...

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Immunology
• Immunology is the scientific study of the immune system and immune responses,
• Also defined as the biochemical study of body defenses against parasites and microbes
(bacteria, virus, fungi and parasites)
• Two major components—innate immunity and adaptive immunity.
• Innate (natural) immunity is the body's first reaction to an invader, is non-specific and quick
response to any sort of pathogen
• Components include physical, cellular, and chemical systems; mucosal barriers, phagocytic
cells, and the action of circulating glycoproteins such as complement
• The adaptive (Acquired) - specific immunity catered against specific antigens.
• Once activated, it triggers the development of new cellular responses and production of
circulating antibody, which have a component of memory if the invader returns.
• Consists of: Humoral immunity, mediated by antibodies and Cell-mediated immunity,
mediated by lymphocytes
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Role of immunology
• Immunodiagnostics provide useful data in their treatment and management
including perspectives on the underlying mechanisms in the disease process.
• Clinical transplantation offers life saving benefits for patients who may need
replacement of organs/tissues,
• Immune based therapies including gene therapy, phototherapy, use of
cytokines and adoptive cell transfer.
• Vaccines – esp in the control of viral and bacterial infections.
• Anthropological studies to determine migration patterns
• Paternity identity and criminal identification through the employment of
DNA and HLA profiles.

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Historical Perspectives
• Immunology as a biological science developed and grew with
increasing understanding of microorganisms
• Developments in Immunology have therefore been associated with
milestones in Microbiology and Parasitology
• Around1000 A.D., ancient Chinese practiced form of immunization by
inhaling dried powders derived from the crusts of smallpox lesions.
• Around fifteenth century, the process of variolation involving applying
powdered smallpox "crusts" and inserting them with a pin or
“poking” device into the skin became commonplace in middle East.

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Historical Perspectives cont’d
• A safer procedure involved substitution of smallpox (variola) virus
with cowpox (vaccinia) virus for the inoculation.
• Cowpox is a benign disease due to infection with vaccinia virus which
is closely related to the virulent variola virus.
• 1774- Benjamin Jesty, an English farmer, inoculated wife with vaccinia
virus = first record using vaccinia virus to "protect" against smallpox.

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Historical Perspectives cont’d
• Later studies in 1796 by Edward Jenner showed that the body could
respond to foreign substances and generate the ability to defend itself
in subsequent infections.
• Exposure to cowpox led to resistance against smallpox.
• Study of disease causing microorganisms later on, thanks to invention
of microscopy led to isolation of various bacteria such as Diphtheria
bacilli.
• The immunity in recovered individuals was found to be induced by
bacterial components and derived products. One of these substances,
diphtheria toxin, was the first antigen to be identified.
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Historical Perspectives cont’d
• The immune response to the foreign substance gave rise to the
definition of an antibody.
• Antibodies were demonstrated in the blood serum or plasma and
found to neutralize bacterial toxins.
• Jules Bordet in 1899 further discovered a serum factor consisting of
several proteins that mediated the destruction of bacteria together
with the antibody- referred to as complement.
• Robert Koch discovered Tubercle bacilli and reported that delayed
type hypersensitivity reaction was responsible for protective
immunity to the bacterial infection that involved white cells.
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Historical Perspectives cont’d
• Louis Pasteur developed the Germ Theory of Disease concerned with
• Prevention of diseases that bacteria caused and how the human body
was changed subsequent to infection so as to resist further insults.
• Virulent chicken cholera bacillus became attenuated by sitting on the
bench over the summer months.
• Similarity between this situation and Jenner’s variolation with vaccinia
virus made him honor Jenner by calling his treatment VACCINATION

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Immunological processes
The body protects against harmful pathogens through two ways:
• Nonspecific host defense mechanism
First line of defense
Second line of defense
• Specific host defense mechanisms or specific immunity. It is also
called the third line defense.

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Definition of terms
• Host defense mechanisms:- are ways in which the human body
protects itself from pathogens.
• Non-specific defense mechanisms:- are ways in which the body
attempts to destroy all types of substances that are foreign to it
including pathogens.
• Specific host defense mechanisms:- antibodies are usually produced
in the body in response to the presence of specific foreign substances
(antigens).

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Components of the immune system
• Consists of the cellular and molecular components derived from the
central and peripheral lymphoid organs.
• Central lymphoid organs
◦consist of the bone marrow and thymus.
◦are the location of maturation of lymphoid cells.
• Peripheral lymphoid organs
◦consist of the spleen, lymph nodes and lymphatic channels, tonsils, adenoids,
Peyer's patches, and appendix.
◦are the location of reactivity of lymphoid cells.

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Components of the immune system
• Cells of the immune system
◦Include the white blood cells (approx. 8000/mm3 of blood), which are
composed of:
◦Granulocytes 50%-80% of white blood cells
◦Lymphocytes 20%-45% of white blood cells
◦Monocytes and macrophages 3%-8% of white blood cells
• Molecules of the immune system
◦Antibodies (immunoglobulins) are protein products of certain lymphocytes
with a precise specificity for a particular antigen.
◦Lymphokines are secreted lymphocyte products that play a role in the
activation of the immune response.

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Innate immunity
• Innate immunity is resistance that exists prior to exposure to the
microbe (antigen).
• It is nonspecific and includes host defenses such as;
o barriers to infectious agents (e.g., skin and mucous membranes),
o certain cells (e.g., natural killer cells),
o certain proteins (e.g., the complement cascade and interferons), and
o processes such as phagocytosis and inflammation
• Innate immunity does not improve after exposure to the organism, in
contrast to acquired immunity, which does.
• Innate immune processes have no memory,
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Innate immunity cont’d
• The innate arm of human host defenses performs two major
functions:
◦Killing invading microbes
◦Activating acquired (adaptive) immune processes.
• Some components of the innate arm, such as neutrophils, only kill
microbes, whereas others, such as macrophages and dendritic cells,
perform both functions, i.e., they kill microbes and present antigen to
helper T cells, which activates acquired immune processes.

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Nonspecific Host Defense Mechanisms
FIRST LINE OF DEFENSE
• These are general and serve to protect the body against many harmful
substances.
a) Mechanical and physical factors
i. Skin:-
• Intact, unbroken skin that covers our bodies serves as a physical or
mechanical barrier to pathogens.
• Very few pathogens are able to penetrate the skin.
• The dryness of most areas of skin inhibits colonization by many pathogens.

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Mechanical and physical factors
ii. Mucous membranes:-
• They serve as physical and mechanical barriers to pathogens.
• Most pathogens can only pass when these membranes have are cut
or scratched.
• The sticky mucous produced by the goblet cells within the mucus
membranes serve to entrap invaders (pathogens).
• They can be removed by normal reflex like coughing or sneezing.
• Such areas include nose (respiratory tract), mouth, and vagina.

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b).Cellular and Chemical factors:-
• Body temperature -<37ºc and acidity of the skin inhibit the growth of
pathogens.
• The oily sebum that is produced by sebaceous glands in the skin
contains fatty acids which are toxic to some pathogens.
• Perspiration (sweat) aids in flushing organisms from pores and surface
of skin.
• Sweat contains enzyme- lysozyme which degrades peptidoglycan in
bacteria cell walls.
• Sloughing off of dead skin cells removes potential pathogens from the
skin.
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Cellular and Chemical factors cont’d
• Sticky mucus produced by the mucous membranes contains lysozyme,
lactoferin and lactoperoxidase that kill bacteria or inhibit growth.
• Lactoferrin binds with iron, a mineral that is required by all
pathogens; because they are unable to compete with lactoferrin for
free iron, the pathogens are deprived of these essential nutrient.
• Lactoperoxidase is an enzyme that produces superoxide radicals,
highly reactive forms of oxygen which are toxic to bacteria.

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Cellular and Chemical factors cont’d
• Because mucosal cells are among the most rapidly dividing cells in the
body, they are constantly being produced and released from the mucous
membranes.
• Bacteria that is adhering to the cells are often expelled along with the
cells they are attached.
• The hair, mucous membranes and irregular chambers of the nose serve
to trap much of the inhaled debris.
• The cilia present on the epithelial cells of the posterior nasal
membranes, nasal sinuses, bronchi and trachea sweep the trapped dust
and microbes upward toward the throat where they are swallowed or
expelled by sneezing and coughing.
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Cellular and Chemical factors cont’d
• Swallowing of saliva can be thought of as a non-specific defense mechanism,
because thousands of bacteria are removed from the oral cavity every time we
swallow.
• Humans swallow approximately 1 litre of saliva per day.
• Digestive enzymes, acidity of the stomach (approx. pH 1.5), alkalinity of the
intestines protects the digestive system from bacteria colonization.
• Peristalsis and expulsion of feces serve to remove bacteria from the intestine.
Bacteria make up 50% of feces.
• Microorganisms are continually flushed from the urethra by frequent urination and
expulsion of mucus secretions.
• The low pH of vaginal fluid usually inhibits colonization of the vagina by pathogens.

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First line of defense cont’d
c) Microbial antagonism:-
• The indigenous microflora prevent colonization by new arrivals to a
particular anatomical site through competition for colonization sites,
nutrients and production of substances that kill other bacteria.

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SECOND LINE OF DEFENSE
• Pathogens able to penetrate the first line of defense are usually
destroyed by non-specific cellular and chemical responses (second
line of defense).
• A complex sequence of events develops involving production of fever,
interferons, activation of the complement system, inflammation,
chemotaxis, and phagocytosis.

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Second line of defense cont’d
a) Transferrin:-
• A glycoprotein synthesized in the liver has a high affinity for iron.
• Its normal function is to store and deliver iron to the host cells.
• Transferrin serves as a nonspecific host defense mechanism by
sequestering iron and depriving pathogens of these essential nutrient.
b) Fever:-
• Normal body temperature fluctuates between 36.20c and 37.50c. Average
37.20c.
• A body temperature greater than 37.80c is generally considered to be
fever.
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Second line of defense cont’d
• Substances that stimulate production of fever are called pyrogens or
pyrogenic substances
• Fever acts as a body nonspecific host defense mechanism by:-
• Stimulating white blood cells (leukocytes) to deploy and destroy
invaders
• Reducing the available free plasma iron, which limits growth of
pathogens that require iron for replication and synthesis of toxins.
• Inducing the production of interleukin-1(IL-1) which causes the
proliferation, maturation, and activation of lymphocytes in the
immunologic response.
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Second line of defense cont’d
c). Interferons:- are small antiviral proteins produced by virus-infected cells .
• They are called interferons because they “interfere” with viral replication.
• There are three types of interferons – alpha, beta and gamma induced by
different stimuli, including viruses ,tumors, bacteria and other foreign cells.
• Alpha-interferon - are produced by B lymphocytes (B cells), monocytes,
macrophages.
• Gamma-interferon - activated by T lymphocytes (T cells) and Natural Killer
cells (NK cells).
• Beta-interferon - by fibroblasts and other virus-infected cells.

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Second line of defense cont’d
• Interferons produced by a virus-infected cell are unable to save that
cell from destruction, but once they are released from that cell, they
attach to the membranes of surrounding cells and prevent viral
replication from occurring in those cells.
• Thus the spread of infection is inhibited, allowing the body to fight
the disease more effectively.

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Second line of defense cont’d
d). The complement system:-
• Is not a single entity, but rather a group of approximately 30 different
proteins (including nine proteins designated as C1-C9) that are found
in normal blood plasma.
• Proteins of the complement system sometimes collectively referred to
as complement components, interact with each other in a step-wise
manner called complement cascade.

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Second line of defense cont’d
• The complement system assists in the destruction of many
different pathogens by
• Initiation and amplification of inflammation
• Attraction of phagocytes to the sites where they are
needed (chemotaxis).
• Activation of leukocytes
• Lysis of bacteria and other foreign cells
• Increased phagocytosis by phagocytic cells (opsonization).

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Second line of defense cont’d
e). Inflammation:-
• The body normally responds to local injury, irritation, microbial invasion or
bacterial toxin by a complex series of events collectively referred to as
inflammation or inflammatory response.
• The 3 major events in acute inflammation are:-
i). An increase in the diameter of capillaries, which increases blood flow
to the site.
ii). Increased permeability of the capillaries, allowing the escape of
plasma and plasma proteins.
iii). Exit of leukocytes from the capillaries and their accumulation at site
of injury.
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Second line of defense cont’d
• The primary purpose of the inflammatory response
-Localize an infection
-Prevent the spread of microbial invaders
-Neutralize any toxins being produced at the site
-Aid in the repair of any damaged tissue

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Second line of defense cont’d
f). Phagocytosis:-
• Is the process by which phagocytes surround and engulf (ingest) foreign material.
• The phagocytic white blood cells are called phagocytes.
• The three major categories of leukocytes found in blood are:
-monocytes
-lymphocytes and
-granulocytes.
• The three types of granulocytes are:-
-eosinophils
-basophils and
-neutrophils.
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Second line of defense cont’d
• The two most important groups of phagocytes in the human body are
macrophages and neutrophils; sometimes called ‛professional
phagocytes ’ because their major function is phagocytosis.
• Phagocytes serve as a ‛clean-up crew’ to rid the body of unwanted
and often harmful substances such as dead cells, unused cellular
secretions, debris and microorganisms.
• Granulocytes –are named from the prominent cytoplasmic granules
that they posses. The phagocytic granulocytes include neutrophils and
eosinophils .
• Neutrophils are much more efficient in phagocytosis than eosinophils
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Second line of defense cont’d
• Macrophages- develop from monocytes during the inflammatory
response to infections.
• Those that leave the blood stream and migrate to infected areas are
called wandering macrophages.
• Fixed macrophages remain in the tissues and organs and serve to trap
foreign debris.
• Macrophages are extremely efficient phagocytes. They are found in
tissues of the reticuloendothelial system(RES).

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Second line of defense cont’d
g). Chemotaxis:-
• Phagocytosis begins when phagocytes move to the site where they are needed.
• The directed migration is called chemotaxis and is the result of chemical
attraction called chemotaxic agents.
• Chemotaxic agents are produced by various cells of the human body are called
chemokines & are produced during the complement cascade and inflammation.
• The phagocytes move along the concentration gradient, meaning they move
from areas of low concentration of chemotaxic agents to the area of the highest
concentration.
• The area of highest concentration is the site where the chemotaxic agents are
being produced or released- often the site of inflammation.
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SPECIFIC (acquired) IMMUNITY
• THIRD LINE OF DEFENSE
• Micro-organisms that overcome non-specific defenses are faced with
specific immunity
• The antigens of the invading micro-organisms comes into contact with
cells of immune system (macrophages and lymphocytes) and initiate a
response
• The response takes two ways:- humoral immunity and cell-mediated
immunity

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 The immune response
1st line 2nd line

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Humoral Immunity
• Humoral (antibody-mediated) immunity is directed primarily against
• Exotoxin-mediated diseases such as tetanus and diphtheria
• Infections in which virulence is related to polysaccharide capsules (e.g
meningococci, pneumococci, haemophilus influenzae).
• Certain viral infections.
• Antibody synthesis is by
• The primary response
• Secondary response
• Antibody response is a physiological reaction to the introduction into
the body of foreign materials, irrespective of whether it is harmful or
not.
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The primary response
• When an antigen is first encountered, antibodies are detectable in the
serum after a long period.
• The lag period is typically 7-10 days but can be longer depending on the
nature and dose of the antigen and the route of administration (oral or
parenteral).
• A small clone of B cells and plasma cells specific for the antigen is formed.
• Serum antibody concentration continues to rise for several weeks, then
drops to low levels.
• The first antibodies formed are IgM, followed by IgG, IgA or both.
• IgM levels decline earlier than IgG levels.
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The secondary response
• When there is a second encounter with the same antigen or closely
related one, months or years after the primary response, there is a rapid
antibody response (in 3-5 days) to higher levels than primary response.
• This is due to persistence of antigen-specific memory cells after the first
contact.
• In the secondary response the amount of IgM produced is qualitatively
similar to that produced during first contact with the antigen;
• However, much more IgG is produced and the levels tend to persist
longer than in the primary response.

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ANTIBODIES
• Antibodies are globulin proteins (immunoglobulins) that react
specifically with the antigen that stimulated their production.
• Are formed by B lymphocytes
• Each individual has a large pool of different B lymphocytes that have a
lifespan of days or weeks and are found in the bone marrow, lymph
nodes, and gut associated lymphoid tissues
• B cells display immunoglobulin on their surface, serving as receptors
for specific antigen
• An antigen interacts with B lymphocyte that shows the best fit as per
its immunoglobulin surface receptor
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Antibody structure

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Antibody structure
• Immunoglobulins are glycoproteins made up of light(L) and heavy(H) polypeptide chains.
• The simplest Ab molecule has a Y shape and consists of 4 polypeptide chains: two H
chains and two L chains. The four chains are linked by disulfide bonds.
• L and H chains are subdivided into variable and constant regions. The regions are
composed of three-dimensionally folded, repeating segments called domains.
• The variable regions of both the light and heavy chain are responsible for antigen-
binding.
• The constant region of the heavy chain is responsible for various biologic functions, e.g.,
complement activation and binding to cell surface receptors.
• The complement binding site is in the CH2 domain.
• The constant region of the light chain has no known biologic function

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Antibodies or Immunoglobulins
IgG:-
• It has two identical antigen binding sites
• Is the predominant antibody in secondary responses and constitutes an
important defence against bacteria and viruses.
• It is the only antibody that can pass the placenta barrier and is the most
abundant immunoglobulin in newborns.
IgM:-
• Is the main immunoglobulin produced early in the primary response
• It is present in the surface of virtually all uncommitted B cells.
• It has ten binding sites
• It is the most efficient immunoglobulin in agglutination complement fixation
and other antigen body reactions
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Antibodies cont’d

IgA:-
• is the main immunoglobulin in secretions such as milk, saliva, and tears
and secretions of the respiratory, intestinal and genital tract.
• It protects the mucous membranes from bacteria and viruses.
IgE:-
• It binds to the receptor on the surface of mast cells, basophils, and
eosinophil. Known for mediating allergic reactions.
IgD:-
• It acts as an antigen receptor when present on the surface of certain B
lymphocytes
• It is present in serum only in trace amounts
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Functions of antibodies
• The most important functions of antibodies are
1) Neutralize toxins and viruses
2) Opsonize microbes so they are more easily phagocytosed.
Opsonization is the process by which antibodies make
microorganisms more easily ingested by phagocytic cells.
3) Activate the complement system,
4) Prevent the attachment of microbes to mucosal surfaces.

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 Antibody Functions

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 Complement System

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Cell-mediated Immunity
• Cell-mediated immunity (CMI) is an immune response that involves the
activation of macrophages and NK-cells, the production of antigen-specific
cytotoxic T-lymphocytes, and the release of various cytokines in response
to an antigen
• This depends on development of lymphoid cells which are specifically
sensitized to the inducing antigen and which react directly with the antigen
to bring about cytotoxic effects
• Development of activated macrophages can also result from this process
• Components: T cells, mФ, NK, cytokines
• Important in intracellular infections such as tuberculosis and viral
infections.
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Role of CMI
• Confer protection against
◦Intracellular protozoa (leishmania, toxoplasma); bacteria
(mycobacteria);
◦Fungi (candida and cryptococci).
• Mediate graft rejections
• Eliminate deleterious autoimmune reactions.
• Inappropriate or exaggerated activation of CMI leads to
o Tissue injury or hypersensitivity disorders
o Granulomatous hypersensitivity reactions and granuloma formation

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Antigen presenting cells
• Immune cell that specialize in presenting an antigen to a T cell,
• An antigen is any substance (immunogen/hapten) foreign to the body
that evokes an immune response alone or after forming a complex with
a larger molecule and that is capable of binding with a product (a T cell
or an antibody) of the immune response.
• Professional APCs : B cells, Macrophages and Dendritic cells and their
derivatives microglial cells, Kupffer's cells, and Langerhans' cells of the
skin
• Non-Professional APCs : Fibroblasts (skin), Thymic epithelial cells,
Thyroid epithelial cells, Glial cells (brain), Pancreatic beta cells, Vascular
endothelial cells
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 Antigen processing

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T Cells
• T cells are one of two primary types of lymphocytes—B cells being the
second type—that determine the specificity of immune response
• Thymus-dependent lymphocytes & develop in the thymus.
• Have a unique antigen receptor of a specific idiotype [T-cell receptor
(TCR)] – distinguishing factor
• Have Fc receptors on some subsets and C3b receptors.
• Develop a series of thymus-induced differentiation markers labeled as
cluster of differentiation (CD) antigens.
• T cells function to actively destroy infected cells, as well as to signal
other immune cells to participate in the immune response.
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TCR and CD Markers
TCR (T Cell Receptor) is the antigen-specific receptor on T cells.
• Consists of a heterodimer
• Most T cells (95%) possess the TCR.
• is associated with CD3.

CD markers
• Arise on T cells during maturation in the thymus.
• Appear on T cells in the following sequence: CD2 (formerly known as
T11), CD3 (T3), CD4 (T4), and CD8 (T8).
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B cells
• B lymphocytes are the cells responsible for antibody responses.
• They develop from precursor cells in the bone marrow before migrating to other lymphoid
tissues but are thymus-independent lymphocytes
• Once activated, these white blood cells produce antibodies.
• B lymphocytes have further roles as antigen-presenting cells and cytokine secretors.
• Classified into four main groups: transitional, naïve, plasma, and memory B cells
• Have a unique surface immunoglobulin (S-Ig) receptor for antigen.
• Develop a series of markers during the differentiation process.
• S-Ig-
• Its the antigen-specific idiotype receptor on B cells.
• Its equivalent to an antibody molecule with a transmembrane projection.
• Its associated with signal transduction molecules, Ig-α and Ig-β.
• Undergoes capping and endocytosis after activation by antigen
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Active immunity
• Is induced after contact with foreign antigens.
• This contact may consist of clinical or sub-clinical infections, immunization with
live or killed infectious agents or antigen exposure to microbial products e.g
toxins, toxoids
• The host actively produces antibodies and lymphocytes acquire the ability to
respond to the antigens.
• Advantages:-
-long-term protection
• Disadvantages:-
-slow onset of protection.
-Need for prolonged or repeated contact with the antigen.
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Active immunity cont’d
Active immunity can be acquired naturally or artificially e.g
• The production of antibodies in response to a pathogen that has
entered the body is an example of natural active acquired immunity.
• The production of antibodies in response to a vaccine is an example of
artificial active acquired immunity.

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Passive immunity
• Is transmitted by antibodies/lymphocytes preformed in another host.
• Advantage:-
-prompt availability of large amounts of antibodies.
• Disadvantages:-
-short lifespan of antibodies
-Possible hypersensitivity reactions if antibodies from
another species are administered.
• Passive immunity can be acquired naturally or artificially

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Passive immunity cont’d
• A foetus receiving antibodies that were produced by the mother is an
example of natural passive acquired immunity.
• E.g. IgG passed from mother to fetus during pregnancy and IgA passed from
mother to newborn during breast feeding

• A soldier receiving antibodies contained in a shot of gamma globulin

is an example of an artificial passive acquired immunity.

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Herd immunity
• When a large proportion of people are immunized in a community,
even those few people who have not been vaccinated also get some
protection because the disease becomes so uncommon.
• This is called herd immunity.
• It is mainly effective for those diseases that pass from man to man e.g
measles, polio and pertusis.

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Immunization
• Is the process of protecting a person from a particular disease.
• It happens when a vaccine against a disease has been given. This is called active
immunization.
• Some vaccines are made from live bacteria or viruses that have been modified
enough not to cause a severe infection, but they are still similar to the original
bacteria or viruses for the body not to be able to make a difference. They are
called live attenuated vaccine.
• Other vaccines are made out of dead bacteria (inactivated) or by modifying the
toxins that some bacteria produce. The modified toxins are called toxoids.
• The vaccines are given either by mouth or injections. They act as antigens.

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Types of vaccines
1. Inactivated Vaccine
2. Attenuated Vaccine
3. Toxoid Vaccine
4. Subunit Vaccine: Subunit vaccines use only part of a target pathogen to provoke a response from the immune
system. This may be done by isolating a specific protein from a pathogen and presenting it as an antigen on its own.
5. Conjugate Vaccine: they’re made using a combination of two different components using pieces from the coats
of bacteria. These coats are chemically linked to a carrier protein, and the combination is used as a vaccine
6. Valence Vaccine: Vaccines may be monovalent. A monovalent vaccine is designed to immunize against a single
antigen or single microorganism. A multivalent or polyvalent vaccine is designed to immunize against two or more
strains of the same microorganism, or against two or more microorganisms.
7. Heterotypic Vaccine: Heterologous vaccines also known as “Jennerian vaccines”, are vaccines that are pathogens
of other animals that either do not cause disease or cause mild disease in the organism being treated.
8. mRNA Vaccine: is a novel type of vaccine which is composed of the nucleic acid RNA, packaged within a vector
such as lipid nanoparticles.

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Major histocompatibility complex (MHC)
• It is a large locus containing closely linked polymorphic genes that code
for cell surface proteins essential for the adaptive immune system.
• Located on the short (p) arm of human chromosome 6.
• Encodes for the major histocompatibility antigens, known in humans as
human leukocyte antigens (HLA).
• Encodes for the class I antigens by the genes HLA-A, HLA-B, & HLA-C.
• Encodes for the class II antigens, defined by the genes in the D (immune
response) region, HLA-DR, HLA-DP, and HLA-DQ.
• Encodes for the class III antigens, which include various complement
components such as C2, C4, factor B, and the C3b receptor.
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• Immunologic role of the histocompatibility antigens:
• Antigens are recognized by the appropriate TCR only in the context of the
histocompatibility antigens.
• Exogenous antigens (e.g, bacteria), which would undergo processing by an APC,
would be expressed on the surface of the APC in the context of a class II MHC
molecule and be recognized by CD4+THcells.
• Endogenous antigens (e.g., virally transformed cell proteins) would be expressed
on the surface of any cell in the context of a class I MHC molecule and be
recognized by a CD8+TC cell.
• Because the MHC gene products, both class I and class II, are cell-surface proteins
and very antigenic, it is important that tissues be typed before transplantation

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• A T-cell-mediated reaction is the main cause of rejection of many types of grafts,
e.g., skin, but antibodies contribute to the rejection of certain transplants, especially
bone marrow
• The acceptance or rejection of a transplant is determined, in large part, by the class
I and class II MHC proteins on the donor cells, with class II playing the major role.
• These alloantigens activate T cells, both helper and cytotoxic, which bear T-cell
receptors specific for the alloantigens. The activated T cells proliferate and then
react against the alloantigens on the donor cells.
• CD8-positive cytotoxic T cells do most of the killing of the allograft cells.
• A graft that survives an acute allograft reaction can nevertheless become
nonfunctional as a result of chronic rejection. This can occur months to years after
engraftment.
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Hypersensitivity reactions
• Hypersensitivity is the term used when an immune response results in exaggerated
or inappropriate reactions harmful to the host.
• The term "allergy" is often equated with hypersensitivity but more accurately should
be limited to the IgE-mediated reactions (Type I: Immediate (Anaphylactic)
Hypersensitivity.)
• The clinical manifestations of these reactions are typical in a given individual and
occur on contact with the specific antigen to which the individual is hypersensitive.
• The first contact of the individual with the antigen sensitizes, i.e., induces the
antibody, and the subsequent contacts elicit the allergic response.
• Hypersensitivity reactions can be subdivided into four main types. Types I, II, and III
are antibody-mediated, whereas type IV is cell-mediated.
• Type I reactions are mediated by IgE, whereas types II and III are mediated by IgG.
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Antigen–Antibody Reactions in the
Laboratory
• An antigen will react only with antibodies elicited by itself or by a
closely related antigen.
• Because of the great specificity, reactions between antigens and
antibodies are suitable for identifying one by using the other. This is
the basis of serologic reactions.
• The results of many immunologic tests are expressed as a titer, which
is defined as the highest dilution of the specimen, e.g., serum, that
gives a positive reaction in the test.

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Type of immunologic diagnostic tests
1. Agglutination
• There are three types of agglutination reactions:
1. Active/Direct agglutination,
2. Passive agglutination and
3. Hemagglutination.
• In this test, the antigen is particulate(e.g. bacteria and red blood cells) or is an
inert particle (latex beads) coated with an antigen.
• Antibody, because it is divalent or multivalent, cross-links the antigenically
multivalent particles and forms a latticework, and clumping (agglutination) can be
seen.
• One very commonly used agglutination test is the test that determines a person's
ABO blood group.
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2. Precipitation (Precipitin)
• In this test, the antigen is in solution.
• The antibody cross-links antigen molecules in variable proportions, and aggregates
(precipitates) form.
• In the zone of equivalence, optimal proportions of antigen and antibody combine;
the maximal amount of precipitates forms, and the supernatant contains neither an
excess of antibody nor an excess of antigen
• In the zone of antibody excess, there is too much antibody for efficient lattice
formation, and precipitation is less than maximal.
• In the zone of antigen excess, all antibody has combined, but precipitation is
reduced because many antigen–antibody complexes are too small to precipitate;
i.e., they are "soluble”
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3. Radioimmunoassay (RIA)
• This method is used for the quantitation of antigens or haptens that can be radioactively
labeled.
• It is based on the competition for specific antibody between the labeled (known) and the
unlabeled (unknown) concentration of material.
• The complexes that form between the antigen and antibody can then be separated and the
amount of radioactivity measured.
• The more unlabeled antigen is present, the less radioactivity there is in the complex. The
concentration of the unknown (unlabeled) antigen or hapten is determined by comparison
with the effect of standards.
• RIA is a highly sensitive method and is commonly used to assay hormones or drugs in serum.
• The radioallergosorbent test (RAST) is a specialized RIA that is used to measure the amount of
serum IgE antibody that reacts with a known allergen (antigen).

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4. Enzyme-Linked Immunosorbent Assay (ELISA)
• This method can be used for the quantitation of either antigens or antibodies in
patient specimens.
• It is based on covalently linking an enzyme to a known antigen or antibody,
reacting the enzyme-linked material with the patient's specimen, and then
assaying for enzyme activity by adding the substrate of the enzyme.
• The method is nearly as sensitive as RIA yet requires no special equipment or
radioactive labels .
• For measurement of antibody, known antigens are fixed to a surface (e.g., the
bottom of small wells on a plastic plate), incubated with dilutions of the patient's
serum, washed, and then reincubated with antibody to human IgG labeled with an
enzyme, e.g., horseradish peroxidase.
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5. Immunofluorescence (Fluorescent Antibody)
• Fluorescent dyes, e.g., fluorescein and rhodamine, can be covalently attached to antibody
molecules and made visible by UV light in the fluorescence microscope.
• Such "labeled" antibody can be used to identify antigens, e.g., on the surface of bacteria
(such as streptococci and treponemes), in cells in histologic section, or in other specimens
• The immunofluorescence reaction is direct when known labeled antibody interacts directly
with unknown antigen and indirect when a two-stage process is used.
• For example, known antigen is attached to a slide, the patient's serum (unlabeled) is added,
and the preparation is washed; if the patient's serum contains antibody against the antigen,
it will remain fixed to it on the slide and can be detected on addition of a fluorescent dye–
labeled antibody to human IgG and examination by UV microscopy.
• The indirect test is often more sensitive than direct immunofluorescence, because more
labeled antibody adheres per antigenic site.
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6. Complement Fixation
• The complement system consists of 20 or more plasma proteins that interact with one another and with cell
membranes.
• Each protein component must be activated sequentially under appropriate conditions for the reaction to progress.
Antigen–antibody complexes are among the activators, and the complement fixation test can be used to identify
one of them if the other is known.
• The reaction consists of the following two steps
(1) Antigen and antibody (one known and the other unknown; e.g., use a known antigen and determine whether a
patient's serum contains antibodies to that antigen) are mixed, and a measured amount of complement (usually
from guinea pig) is added. If the antigen and antibody match, they will combine and use up ("fix") the complement.
(2) An indicator system, consisting of "sensitized" red blood cells (i.e., red blood cells plus anti–red blood cell
antibody), is added. If the antibody matched the antigen in the first step, complement was fixed and less (or none) is
available to attach to the sensitized red blood cells. The red blood cells remain unhemolyzed; i.e., the test is positive,
because the patient's serum had antibodies to that antigen. If the antibody did not match the antigen in the first
step, complement is free to attach to the sensitized red blood cells and they are lysed; i.e., the test is negative.

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7. Neutralization Tests
• These use the ability of antibodies to block the effect of toxins or the infectivity of viruses. They can be used
in cell culture (e.g., inhibition of cytopathic effect and plaque-reduction assays) or in host animals (e.g.,
mouse protection tests).

8. Immune Complexes
• Immune complexes in tissue can be stained with fluorescent complement. Immune complexes in serum can
be detected by binding to C1q or by attachment to certain (e.g., Raji lymphoblastoid) cells in culture.

9. Hemagglutination Tests
• Many viruses clump red blood cells from one species or another (active hemagglutination). This can be
inhibited by antibody specifically directed against the virus (hemagglutination inhibition) and can be used to
measure the titer of such antibody. Red blood cells also can absorb many antigens and, when mixed with
matching antibodies, will clump (this is known as passive hemagglutination, because the red cells are passive
carriers of the antigen).
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10. Antiglobulin (Coombs) Test
• Some patients with certain diseases, e.g., hemolytic disease of the newborn (Rh
incompatibility) and drug-related hemolytic anemias, become sensitized but do not exhibit
symptoms of disease. In these patients, antibodies against the red cells are formed and bind
to the red cell surface but do not cause hemolysis
• These cell-bound antibodies can be detected by the direct antiglobulin (Coombs) test, in
which antiserum against human immunoglobulin is used to agglutinate the patient's red
cells.
• In some cases, antibody against the red cells is not bound to the red cells but is in the serum
and the indirect antiglobulin test for antibodies in the patient's serum should be performed.
• In the indirect Coombs test, the patient's serum is mixed with normal red cells and antiserum
to human immunoglobulins is added. If antibodies are present in the patient's serum,
agglutination occurs.

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11. Western Blot
• This test is typically used to determine whether a positive result in a
screening immunologic test is a true-positive or a false-positive result.
• For example, patients who are positive in the screening ELISA for HIV
infection or for Lyme disease should have a Western blot test
performed.

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12. Flow Cytometry
• Also called Fluorescence-Activated Cell Sorting
• This test is commonly used to measure the number of the various types of
immunologically active blood cells
• For example, it is used in HIV-infected patients to determine the number of CD4-
positive T cells.
• In this test, the patient's cells are labeled with monoclonal antibody to the
protein specific to the cell of interest, e.g., CD4 protein if the number of helper T
cells is to be determined. The monoclonal antibody is tagged with a fluorescent
dye, such as fluorescein or rhodamine. Single cells are passed through a laser
light beam, and the number of cells that fluoresce is counted by use of a machine
called a fluorescence-activated cell sorter (FACS).
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Antigen–Antibody Reactions Involving Red
Blood Cell Antigens
• Many different blood group systems exist in humans.
• Each system consists of a gene locus specifying antigens on the erythrocyte surface. The two most
important blood groupings, ABO and Rh, are described below.

• The ABO Blood Groups & Transfusion Reactions

• All human erythrocytes contain allo antigens (i.e., antigens that vary among individual members of a
species) of the ABO group. This is an important system, which is the basis for blood-typing and
transfusions.
• The A and B genes encode enzymes that add specific sugars to the end of a polysaccharide chain on
the surface of many cells, including red cells
• People who inherit neither gene are type O. The genes are codominant, so people who inherit both
genes are type AB.
• People who are either homozygous AA or heterozygous AO are type A, and, similarly, people who are
either homozygous BB or heterozygous BO are type B.
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Compatibility of blood types

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Rh Blood Type & Hemolytic Disease of the
Newborn
• About 85% of humans have erythrocytes that express the Rh(D) antigen, i.e., are Rh(D)+.
• When a Rh(D)–person is transfused with Rh(D)+ blood or when an Rh(D)–woman has an Rh(D)+fetus
(the D gene being inherited from the father), the Rh(D) antigen will stimulate the development of
antibodies.
• This occurs most often when the Rh(D)+ erythrocytes of the fetus leak into the maternal circulation
during delivery of the first Rh(D)+ child.
• Subsequent Rh(D)+pregnancies are likely to be affected by the mother's anti-D antibody, and
hemolytic disease of the newborn (erythroblastosis fetalis) often results.
• This disease results from the passage of maternal IgG anti-Rh(D) antibodies through the placenta to
the fetus, with subsequent lysis of the fetal erythrocytes.
• The direct antiglobulin (Coombs) test is typically positive.
• The problem can be prevented by administration of high-titer Rh(D) immune globulins (Rho-Gam) to
an Rh(D)–mother at 28 weeks of gestation and immediately upon the delivery of an Rh(D)+child. These
antibodies promptly attach to Rh(D)+erythrocytes and prevent their acting as sensitizing antigen. This
prophylaxis is widely practiced and effective.
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Immunological tolerance

• Immunological tolerance is a complex series of mechanisms that

impair the immune system to mount responses against self antigens.
• Central tolerance occurs when immature lymphocytes encounter self
antigens in the primary lymphoid organs, and consequently they die
or become unreactive.
• Peripheral tolerance occurs when mature lymphocytes, escaped from
negative selection during ontogeny, encounter self antigens in
secondary lymphoid organs and undergo anergy, deletion or
suppression.

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Immunological tolerance cont’d
• A heterogeneous family of T regulatory cells has recently been identified,
which have been found to play an important role in suppressing immune
responses against self.
• Failure or breakdown of immunological tolerance results in
autoimmunity and autoimmune diseases.
• Such events are related to both genetic and environmental factors, the
latter being mainly represented by infections.
• Infectious agents can indeed promote autoimmune responses either by
inducing tissue inflammation and therefore an unintended bystander
activation of autoreactive T cells, or by promoting T cell responses to
microbial epitopes that cross react against self peptides.
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Autoimmune diseases
• In certain circumstances tolerance may be lost and immune reactions
to host antigens may develop, resulting in autoimmune diseases.
• The most important step in the production of autoimmune disease is
the activation of self-reactive helper (CD4) T cells.
• These self-reactive Th-1 or Th-2 cells can induce either cell-mediated
or antibody-mediated autoimmune reactions, respectively.
• Autoimmunity, thus is the state in which the immune system reacts
against the body’s own normal components, producing disease or
functional changes.

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• Many autoimmune diseases exhibit a marked familial incidence,
which suggests a genetic predisposition to these disorders.
• There is a strong association of some diseases with certain human
leukocyte antigen (HLA) specificities, especially the class II genes.
• For example, rheumatoid arthritis occurs predominantly in individuals
carrying the HLA-DR4 gene. Ankylosing spondylitis is 100 times more
likely to occur in people who carry HLA-B27, a class I gene, than in
those who do not carry that gene.

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Tumor immunology
• The innate and adaptive immune systems function to protect the host from
foreign pathogens, and are generally tolerant toward host tissues –
adequately differentiating between “self” and “non-self” antigens.
• In the setting of an evolving tumor, the immune system is likely exposed to
numerous, previously unseen, antigens arising from genetic abnormalities.
• The immune system is able to perceive and eliminate some tumors early on
in their development.
• However, the theory of immunoediting, which involves the process of
immunosurveillance, suggests that certain tumors escape from an
equilibrium state previously held in check by the immune system, and
become clinically significant.
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Tumor immunosurveillance:-
• First, the immune system by eliminating or suppressing viral
infections, protects the host from tumors induced by viruses
• Second, by the elimination of pathogens and prompt resolution of
inflammation, it prevents the establishment of an inflammatory
environment that is conducive to tumorigenesis
• Third, the immune system could specifically identify and eliminate
tumor cells by the expression of tumor-specific antigens or molecules
that are induced by cellular stress

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• Mechanisms responsible for immune cell dysfunction in cancer are
numerous.
• The escape from tumor immunity can be achieved in three principally
distinct ways:
1. Lack of recognition by means of loss or alteration of molecules which are
important for the recognition and activation of the immune system
2. Lack of susceptibility, i.e., escape from the effector mechanisms of cytotoxic
lymphocytes
3. Induction of immune dysfunction

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• Questions?

• Clarification?

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 REFERENCES
• Sherri’s Medical Microbiology 7th edition
• Immunology by A.K. Chemtai, 3rd edition; JKF 2006

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 THANK YOU!

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