SE "Dnipropetrovsk Medical

Academy"

The principles of the immune

system, clinical and laboratory
evaluation of its disorders
 What is immunology ?
Immunology is a branch of biomedical science
that covers the study of all aspects of the immune
system in all organisms

The study of all aspects of the immune system,

including its structure and function, disorders of
the immune system, blood banking, immunization,
and organ transplantation
 Clinical immunology
 Clinical immunology is the study of diseases caused
by disorders of the immune system (failure, aberrant
action, and malignant growth of the cellular elements
of the system). It also involves diseases of other
systems, where immune reactions play a part in the
pathology and clinical features.
 Clinical immunologists also study ways to prevent
transplant rejection, in which the immune system
attempts to destroy allografts.
Pathogens representing the major categories
of microorganisms causing human diseases
THE NATURE OF ANTIGENS
Historically named as antibody generators
– Molecule which stimulates production of and
binds specifically to an antibody
Contemporary view distinguishes between
– Antigen
• Molecule which can bind to specific antibody but cannot
elicit adaptive immune response
– Immunogen
• Molecule which can stimulate adaptive immune response

Best immunogens are proteins with

MW > 10,000
THE NATURE OF ANTIGENS
 Carbohydrates, nucleic acids and lipids are
also potential antigens / immunogens
 Hapten
– Small (low MW) molecule unable to elicit
immune response
– Combines with larger carrier molecule which
together function as immunogen
– Antibody may react independently with hapten
following hapten/carrier adaptive immune
response
– Example
• Penicillin G (MW of 372)
• Albumin (MW of 66,000)
 Immune System: organs
CENTRAL:
 Thymus
 Bone marrow

PERIPHERAL:
 Spleen
 Tonsils and adenoids
 Lymph nodes
 Payer’s patches
 Appendix
 Lymphatic vessels
 MALT, GALT etc.
 THE LYMPHATIC SYSTEM
 Lymph
– Fluid and cells in lymphatic vessels

 Lymphatic vessels
– Collect and return interstitial fluid to blood
– Transport immune cells throughout body
– Transport lipid from intestine to blood

 Lymph nodes
– Kidney shaped organs at intervals along lymphatic vessels

 Other secondary lymphatic tissues and organs

 LYMPHOCYTES AND THE
LYMPH NODES
 Naive lymphocytes circulate between blood,
lymph and secondary lymph nodes
 Pathogens from infected tissue sites are picked
up by lymphatic vessels and arrive at closest
lymph node
 T and B cells congregate at specific regions of
nodes
 Architecture and size of nodes change in
response to activation of lymphocytes
Lymphnodes
 LYMPHOCYTES AND THE SPLEEN

 Spleen
– Lymphoid organ in upper left abdomen
– Functions
• Remove damaged or old erythrocytes
• Activation of lymphocytes from blood borne pathogens

 Architecture of Spleen
– Red pulp
• Erythrocytes removed
– White pulp
• Lymphocytes stimulated
THE SPLEEN
 SECONDARY LYMPHOID TISSUES
ASSOCIATED WITH MUCOUS MEMBRANES

 Primary portals of entry for pathogens

– Respiratory tract
– Gastrointestinal tract

 Secondary lymphoid tissues

– Bronchial-associated lymphoid tissue (BALT)
– Gut-associated lymphoid tissues (GALT)
• Tonsils, adenoids, appendix, Peyer’s patches

 Pathogens are directly transferred across

mucosa by “M” cells
Gut-associatedlymphoid tissues
 Immune system: cells
 Lymphocytes
– T-lymphocytes
– B-Lymphocytes, plasma cells
– natural killer lymphocytes
 Monocytes, Macrophage
 Granulocytes
– neutrophils
– eosinophils
– basophils
 ORIGIN OF CELLS OF THE
IMMUNE SYSTEM
 Derived from common progenitor cell
in bone marrow
– Pluripotent hematopoietic stem cell

 Progenitor Stem Cells

– Erythroid lineage
• Erythrocytes and Megakaryocytes
– Myeloid lineage
• Monocyte/macrophage, dendritic cells, PMN’s, mast
cells
– Lymphoid lineage
• Small and large lymphocytes
CELLS OF THE IMMUNE SYSTEM
Lymphocytes
Resting
lymphocyte
 T and B cells - electron
microscope
Resting lymphocyte
Activated
B cell

Alberts et al.

Activated T cell
 CELLS OF INNATE IMMUNITY

 Myeloid Lineage
– Neutrophil
• Principal phagocytic cell of innate immunity
– Eosinophil
• Principal defender against parasites
– Basophil
• Functions similar to eosinophils and mast cells
– Referred to as
• Polymorphonuclear leukocytes (PMN’s)
– Nuclei are multilobed (2 to 5)
• Granulocytes
– Cytoplasmic granules
Myeloid Lineage

http://www.lab.anhb.uwa.edu.au/mb140/CorePages/Blood/blood.htm
Myeloid Lineage

http://www.lab.anhb.uwa.edu.au/mb140/CorePages/Blood/blood.htm
http://www.lab.anhb.uwa.edu.au/mb140/CorePages/Blood/blood.htm
 CELLS OF INNATE AND ADAPTIVE
IMMUNITY
 Myeloid lineage
– Monocytes
• Leukocytes with bean shaped or brain-like
convoluted nuclei
• Circulate in blood with half life of 8 hours
• Precursors of tissue macrophages
– Macrophages
• Mononuclear phagocytic cells in tissue
• Derive from blood monocytes
• Participate in innate and adaptive immunity
CELLS OF INNATE AND ADAPTIVE IMMUNITY
Monocyte-Macrophage Cell System
 Macrophages participate in phagocytosis, inflammation, and
cellular immunity

 Macrophages are mainly involved in nonspecific immunity and

include the phagocytic cells: mononuclear phagocytes,
polymorphonuclear phagocytes (neutrophils), eosinophils and
mediator cells: basophils, mast cells and platelets

Macrophage - scanning
electron microscope

Monocyte
 CELLS OF INNATE AND ADAPTIVE
IMMUNITY
 Myeloid lineage
– Dendritic cells
• Cells with dendriform (star shaped)
morphology
• Interdigitating reticular cells (synonym)
• Capture and present antigens to T
lymphocytes
– Mast cells
• Located in mucous membrane
and connective tissue throughout body
• Major effector cell in allergy
• Modulation of initial immune response
Myeloid lineage
 Immune system: molecules

 Antibodies
 Complement
 Cytokines
 Interleukines
 Interferons
 CLASSIFICATION OF ANTIBODIES
(IMMUNOGLOBULINS)
 Five (5) classes (isotypes)
– Immunoglobulin A (IgA)
– Immunoglobulin G (IgG)
– Immunoglobulin M (IgM)
– Immunoglobulin D (IgD)
– Immunoglobulin E (IgE)

 Based on structural differences in constant

regions of heavy chains
 Classes have specialized effector functions
ANTIBODIES (IMMUNOGLOBULINS)
ANTIBODIES (IMMUNOGLOBULINS)
 IMMUNOGLOBULINS IgG

 70-75% of total immuniglobulin. Y-shaped molecule

having 2 light chains and 2 Gamma heavy chains
 4 subclasses which differ in their heavy chain
composition and in some of their characteristics such
as biologic activities IgG1, IgG2, IgG3 and IgG4
 Secreted in high quantities in secondary exposures
 Ability to cross the placenta, maternal IgG provides
the major line of defense against infection for the
first few weeks of a baby's life
 The serologic behavior and characteristics of IgG
antibodies make them one of the most clinically
significant in blood banking
 IMMUNOGLOBULINS IgG
 Most blood group antigens capable of eliciting
an immune response result in the production of
IgG antibodies.
• These antibodies are detected by serologic test procedures
based on their behavior characteristics, such as reactivity at 37
C, complement activation, indirect agglutination and hemolysis.
• Much of routine blood banking involves serologic test procedures
designed to detect and identify IgG antibodies.

 Major functions / applications

• neutralize microbes and toxins
• opsonize antigens for phagocytosis
• activate the complement
• protect the newborn
 IMMUNOGLOBULINS IgM
 Largest of all the antibody molecules and
the structure consists of five of the basic
units (pentamer) joined together by a
structure known as J-chain.
 Accounts for about 5-10% of the immunoglobulin pool.
 Restricted almost entirely to the intravascular space due to
its large size.
 Fixes complement and is much more efficient than IgG in
the activation of complement and agglutination.
 First antibody to be produced and is of greatest importance
in the first few days of a primary immune response to an
infecting organism. does not cross the placenta.
 IMMUNOGLOBULINS IgM
 Many blood group antibodies that are
capable of agglutinating antigen positive
RBCs suspended in saline in tests
performed at 22 C are IgM causing visible
agglutination, ie, ABO antibodies
 IgM antibodies are potent agglutinators that activate
complement very efficiently
 Major functions / applications
• secreted first during primary exposure
• activates the complement
• used as a marker of recent
 IMMUNOGLOBULINS IgA

 Represents 10 to 15% of the total circulatory

immunoglobulin pool
 The dimeric IgA molecule, monomeric in serum
 Found in saliva, tears, colostrum breast milk and in nasal,
bronchial and intestinal secretions. IgA is present in large
quantities in colostrum and breast milk and can be
transferred across the gut mucosa in the neonate and plays
an important role in protecting the neonate from infection
 Produced in high concentrations by lymphoid tissues lining
the gastrointestinal, respiratory and genitourinary tracts and
protection it against infections and preventing absorption of
potential antigens in the food we eat
 IMMUNOGLOBULINS IgA
 In plasma IgA may exist as a single basic
structural unit or as two or three basic
units joined together
 The IgA present in secretions exists as 2 basic units (a
dimer) attached to another molecule know as secretory
component
• This substance is produced by the cells lining the
mucous membranes
• It is thought to protect the IgA in secretions from
destruction by digestive enzymes
 IgA does not cross the placenta and does not bind
complement
 IMMUNOGLOBULINS IgD
 Accounts for less than 1% of the total
immunoglobulin pool. Monomeric
 This is primarily a cell membrane
immunoglobulin found on the surface of B
lymphocytes
 IgD does not fix complement and does not cross the
placenta
 Little is known about the function of this class of antibody.
 No blood group antibodies have been reported to belong
to this class
 IMMUNOGLOBULINS IgD

 Major functions / applications

– present on the surface of B lymphocytes
– functions as membrane receptor
– role unclear
• has a role in antigen stimulated lymphocyte
differentiation
 IMMUNOGLOBULINS IgE
 Monomeric, mediates type I hypersensitivity
 Trace plasma protein (only about 0.004%)
in the plasma of non-parasitized individuals.
 Major importance because it mediates
some types of allergic reactions and is
generally responsible for an individual's
immunity to invading parasites
 Fc region binds strongly to a receptor on mast cells and
basophils and, when antigen is bound it causes the basophil (or
mast cell) to release histamines and heparin from these cells,
resulting in allergic symptoms
 IMMUNOGLOBULINS IgE
 Clinical effects of IgE mediated
reactions include increased vascular
permeability, skin rashes, respiratory
tract constriction (wheezing), and
increased secretions from epithelium
(watery eyes, runny nose)
 Not much else is known about its biologic role
 IgE does not fix complement and does not cross the
placenta
 No blood group antibodies have been reported to belong to
this class
 Complement
 Three primary functions:
– Lysis of antibody coated cells, such as bacteria
and RBCs
– Mediation of opsonization, preparation of
foreign cells for phagocytosis
– Generation of peptide fragments that regulate
features of the inflammatory and immune
response
Comparison of 3 Pathways
 The Classic Pathway
 Eleven components involved, numbered C1 to
C9 Two IgG, One IgM

 Complement cascade requires presence of

cations, both calcium and magnesium

 Activation of the classic pathway almost

always initiated by immunoglobulin

– Requires only 1 molecule of IgM

– Requires 2 molecules of IgG

 Alternative (Properdin) Pathway
 Proteins in the alternative pathway perform activities
similar to those in the classic pathway but are usually non-
antibody triggered
 Any one of a variety of substances can initiate complement
activation including:
– bacterial polysaccharides and lipopolysaccharides,
– endotoxins,
– cobra venom,
– trypsin like enzymes,
– and aggregates of IgA and IgG4 that do not activate C1.
 C1, C4 and C2 do not participate
 Alternative pathway: C3,C5,C6,C7,C8,C9
Alternative Pathway
 Lectin Pathway
 Activation begins when mannan-binding protein (MBP)
binds to the mannose groups of the microbial
carbohydrates
 Two more lectin pathway proteins called MASP1 and
MASP2 (equivalent to C1r and C1s of the classical
pathway) now bind to the MBP
 This forms an enzyme similar to C1 of the classical
complement pathway that is able to cleave C4 and C2 to
form C4bC2a, the C3 convertase capable of
enzymatically splitting hundreds of molecules of C3 into
C3a and C3b
 Lectin Pathway
 The beneficial results are the same as in
the classical complement pathway above:
– trigger inflammation (C5a>C3a>c4a)
– chemotactically attract phagocytes to the infection site
(C5a)
– promote the attachment of antigens to phagocytes via
enhanced attachment or opsonization (C3b>C4b)
– serves as a second signal for the activation of naive B-
lymphocytes (C3d)
– cause lysis of gram-negative bacteria and human cells
displaying foreign epitopes (MAC)
– and remove harmful immune complexes from the body
(C3b>C4b)
 Regulation of Complement
 Activation of complement cascade results in complex series
of molecular event with potent biologic consequences
 Modulating mechanisms are necessary to regulate
complement activation and control production of biologically
active split products
 First mechanism is spontaneous decay of activated
components
 Second mechanism involves specific control proteins that
modulate the activity of certain complement components at
critical activation steps
– C1 inhibitor blocks activities of C1r and C1s.
– Other factors inhibit activation of other complement components.
 A number of proteins act to control the membrane attack unit
 THE IMMUNE RESPONSE AND
IMMUNITY
 Immune response
– Innate (non-specific)
– Adaptive (specific)
• Primary
• Secondary

 Immunity
– State of non-specific and specific protection

 Acquisition of Immunity
– Natural
– Artificial
 Immune system

Acquired Innate
immune system immune system

Humoral Cellular 1st line of 2nd line of

(antibody mediated) (Cell mediated) defense defense
immune response immune response

Skin
Mucous Inflammation
B-lymphocytes T-lymphocytes
membrane Phagocytosis
Complement
Interferons
Natural killers
 Types of immunity

 Innate (non-adaptive)
– first line of immune response
– relies on mechanisms that exist before infection

 Acquired (adaptive)
– Second line of response (if innate fails)
– relies on mechanisms that adapt after infection
– handled by T- and B- lymphocytes
– one cell determines one antigenic determinant
Immune system

Non-specific Specific
• Neutrophils
• Macrophages • Lymphocytes
• Complement
• Mechanical

“INNATE” “ADAPTIVE”
 Defense Against Disease

Nonspecific External Barriers

skin, mucous membranes

If these barriers are penetrated,

the body responds with

Innate Immune Response

phagocytic and natural killer cells, inflammation, fever

If the innate immune response is insufficient, the body responds with

Adaptive Immune Response

cell-mediated immunity, humoral immunity
Innate immunity (non-adaptive)
 Based on genetic make-up
 Relies on already formed components
 Rapid response: within minutes of
infection
 Not specific
– same molecules / cells respond to a range of
pathogens
 Has no memory
– same response after repeated exposure
 Does not lead to clonal expansion
 Innate immunity: mechanisms
 Mechanical barriers / surface secretion
– skin, acidic pH in stomach, cilia
 Humoral mechanisms
– lysozymes, basic proteins, complement, interferons
 Cellular defense mechanisms
– natural killer cells neutrophils, macrophages, mast cells,
basophils, eosinophils

NK Cell Eosinophils Monocyte

Neutrophil Basophils &
Mast cells Macrophage
 First line of defense
Non-specific defenses are designed to
prevent infections by viruses and bacteria.
These include:
– Intact skin
– Mucus and Cilia
– Phagocytes
The first lines of defense (Anatomical barriers):
The natural barriers

1- the skin 2- mucous membranes

That protect organ system open to the external environment such as the
digestive, respiratory, reproductive system and urinary system

Dr. Sanaa Tork

 Component Functions
Skin and mucous membranes – mechanical factors
Intact skin Forms a physical barrier to the entrance of microbes
Inhibit the entrance of many microbes, but not as
Mucous membranes
effective as intact skin
Mucus Traps microbes in respiratory and digestive tracts

Hairs (nostril hairs) Filter incoming air from microbes and dust in nose

Cilia (on cells lining the Together with mucus, trap and remove microbes
respiratory tract) and dust from upper respiratory tract

Tears dilute and wash away irritating substances

Tear ducts
and microbes
Washes microbes from surfaces of teeth and
Saliva
mucous membranes of mouth
Epiglottis Prevents microbes and dust from entering trachea

Urine Washes microbes from urethra

 Component Functions
Skin and mucous membranes – chemical factors
Gastric juice Destroys bacteria and most toxins in stomach
Acid pH of skin Discourages growth of many microbes
Unsaturated fatty acids Antibacterial substance in sebum (sweat)
Defensins (low found in the lung and gastrointestinal tract have
molecular weight antimicrobial activity
proteins)
in the lung (substances that promote phagocytosis
Surfactants
of particles by phagocytic cells)
found in tears, saliva and nasal secretions can
Lysozyme and
breakdown the cell wall of bacteria and destabilize
phospholipase
bacterial membranes (Antimicrobial substance)
Skin and mucous membranes – biological factors
On the skin and in the gastrointestinal tract can
prevent the colonization of pathogenic bacteria by
The normal flora secreting toxic substances or by competing with
pathogenic bacteria for nutrients or attachment to
cell surfaces
 Role of phagocytes
 Phagocytes are several types
of white blood cells (including
macrophages, dendritic cells,
eosinophils, neutrophils) that
seek and destroy invaders.
Some also destroy damaged
body cells
 Phagocytes are attracted by
an inflammatory response of
damaged cells
 Phases of phagocytosis
1. Chemotaxis and adherence of microbe to

phagocyte

2. Attachments and uptake (Ingestion of microbes by

phagocyte

3. Digestion

4. After ingesting a foreign invader

 Phases of phagocytosis
1. Chemotaxis and adherence of microbe to phagocyte:
when pathogens invade the body and cause an infection, some of the
cells under attack respond by releasing chemicals such as histamine.
These with any chemicals released by the pathogens themselves, attract
passin phagocytes to the site. The phagocytes destroy the pathogens by
phagocytosis.

2. Attachments and uptake (Ingestion of microbes by

phagocyte): The phagocytes move towards the pathogens, which
may be clustered together and covered in antibodies. This further
stimulates the phagocytes to attack them. This is because phagocytes
have receptor proteins on their surfaces that recognize antibody
molecules and attach to them.
 Phases of phagocytosis
3. Digestion:
formation of phagosome, fusion of phagosome with lysosyme to form
phagolysosome, digestion of the ingested microbes and the formation
of residual body

4. After ingesting a foreign invader

they “wear” pieces of it called antigens (antigenic determinant) on their
cell membrane receptors (Major Histocompatibility Complex (MHC) –
in this case it named Antigen Presenting Cell (APC) this tells other
types of immune system cells what to look for
Phases of phagocytosis
Second line of innate defence
Complement:
 Complement is not a cell but a group of proteins
 Set of about 30 different kinds of proteins that circulate in an inactive form
in the blood
 These proteins can act together (in complement) with other defense
mechanisms
 Made in the liver
 Activated by infection: Substances on the surfaces of many microbes
trigger a cascade of steps that activate the complement system, leading
to the lyses (bursting) of the invaders, activated complements:
– help to recruit phagocytes to site of inflammation and activate them
– bind to receptors on phagocytes, helping to remove agent of infection
– form pores in the invader or infected cell’s membrane (like the NKs do)
– activate mast cells to release histamine and other factors
– Certain complement proteins help trigger the inflammatory responses
Second line of innate defence Complement:
 Natural killer cells (NK cells)
 instead of attacking the invaders, they attack
the body’s own cells that have become
infected by viruses
 they also attack potential cancer cells, often
before they form tumors
 they bind to cells using an antibody “bridge”,
then kill it by secreting a chemical (perforin)
that makes holes in the cell membrane of the
target cell. With enough holes, the cell will
die, because water rushing inside the cell will
induce osmotic swelling, and an influx of
calcium may trigger apoptosis
 Mast cells
 found in tissues (connective tissues and mucous
membranes) like the skin, near blood vessels

 are activated after antigen binds to a specific type

of antibody called IgE that is attached to receptors
on the mast cell

 activated mast cells release substances that

contribute to inflammation, such as histamine

 mast cells are important in allergic responses but

are also part of the innate immune response,
helping to protect from infection
Soluble factors
 Interferon
Interferon is a protein produced by virus-infected cells that inhibits
the synthesis of viral proteins, leading to decreased viral
replication. It also can cause apoptosis of virus-infected cells

 Acute phase proteins

– proteins in the plasma that increase during infection and
inflammation
– can be used diagnostically to give an indication of acute
inflammation
– an example of an acute phase protein is C-reactive protein,
which fixes complement.
 Adaptive immunity
second line of response
 Based upon resistance acquired during life
 Relies on genetic events and cellular growth
 Responds more slowly, over few days
 Is specific
– each cell responds to a single epitope on an antigen
 Has anamnestic memory
– repeated exposure leads to faster, stronger response
 Leads to clonal expansion
NATURALLY ACQUIRED IMMUNITY
 Active
– Antigens enter body naturally with response of
• Innate and adaptive immune systems
– Provides long term protection

 Passive
– Antibodies pass from mother to
• Fetus across placenta
• Infant in breast milk
– Provides immediate short term protection
NATURALLY ACQUIRED IMMUNITY
Active Passive
Immunity Immunity
Natural clinical, sub- via breast milk,
clinical infection placenta

Artificial Vaccination: immune serum,

immune cells
Live, killed,
purified antigen
vaccine
Adaptive immunity: mechanisms
 Cell-mediated immune response (CMIR)
– T-lymphocytes
– eliminate intracellular microbes that survive within
phagocytes or other infected cells
 Humoral immune response (HIR)
– B-lymphocytes
– mediated by antibodies
– eliminate extra-cellular
microbes and their toxins

Plasma cell
(Derived from B-lymphocyte,
produces antibodies)
 Cell-mediated immune response
1. T-cell
– recognizes peptide antigen
on macrophage in
association with major
histo-compatibility
complex (MHC) class
– identifies molecules on
cell surfaces
– helps body distinguish self
from non-self
2. T-cell goes into effectors
cells stage that is able to
kill infected cells
 Cell mediated immune response
 Primary response
– production of specific clones of effector T cells and memory
clones
– develops in several days
– does not limit the infection
 Secondary response
– more pronounced, faster
– more effective at limiting the infection

Example - cytotoxic reactions against intracellular parasites, delayed

hypersensitivity (e.g., Tuberculin test) and allograft rejection.
 T lymphocytes
and cell-mediated immunity
 Originate from stem cells in bone marrow
followed by migration to thymus gland
 Maturation takes place in thymus gland followed
by migration to secondary lymphoid tissue
 Respond to antigens on the surface of antigen
presenting cells (APC’s)
 Antigen presenting cells (APC’s)
– Macrophages
– Dendritic cells
– B lymphocytes
 T lymphocytes
and cell-mediated immunity
 Antigen presenting cells (APC’s)
– Ingest and process antigens then display fragments
(short peptides) on their surface in association with
molecules of major histocompatibility complex (MHC)
 Major histocompatibility (MHC) molecules
– MHC class I molecules
• Present antigens to CD8 T cells
– MHC class II molecules
• Present antigens to CD4 T cells
 T cells which encounter antigen differentiate into
effector T cells
Major histocompatibility (MHC) molecules
Major histocompatibility (MHC) molecules
Major histocompatibility (MHC) molecules
Immune response
THE CLUSTER OF DIFFERENTIATION (CD)
 A protocol for identification and
investigation of cell surface molecules
 CD number assigned on basis of 1 cell
surface molecule recognized by 2 specific
monoclonal antibodies
 CD nomenclature established in 1982
– 1st International Workshop and Conference on
Human Leukocyte Differentiation Antigens (HLDA)
THE CLUSTER OF DIFFERENTIATION (CD)

Cells CD markers on leukocytes

Granulocyte CD45+, CD15+
Monocyte CD45+, CD14+
T lymphocyte CD45+, CD3+
T helper lymphocyte CD45+, CD3+, CD4+
T cytotoxic lymphocyte CD45+, CD3+, CD8+
B lymphocyte CD45+, CD19+, CD20+, CD22+,
Natural killer cell CD45+, CD16+, CD56+, CD3-
 Roles of effector T cells
in immune response
 CD8 cytotoxic T cells
– enter bloodstream and travel to infection site
– kill cells infected with viruses and other
intracellular microorganisms
 CD4 TH1 helper T cells
– enter blood stream and travel to infection site
– help activate macrophages
 CD4 TH2 helper T cells
– work within secondary lymphoid tissues
– help activate B cells
 Humoral Immune Response
 Production of antibodies induced when the
host's immune system comes into contact
with foreign antigenic substance and reacts
to this antigenic stimulation.
 Two types of responses:
– primary
– secondary
 Humoral Immune Response
 Antibody production occurs in four phases
following antigen challenge:
– lag phase when no antibody is detectable
– log phase in which antibody titer rises
logarithmically
– plateau phase during which the antibody titer
remains steady
– decline phase during which antibody levels
gradually decline
 Humoral Immune Response
 You must be able to differentiate a primary vs secondary
immune response based on the following:
‒ Time
‒ Antibody Titer
‒ Antibody Class
‒ Antibody affinity and avidity
 These are critical to understanding reactions obtained in
Blood Banking
 The following chart nicely illustrates the concepts.
 Humoral Immune Response
1. B lymphocytes recognize
specific antigens
– proliferate and
differentiate into
antibody-secreting
plasma cells
2. Antibodies bind to specific
antigens on microbes;
destroy microbes via
specific mechanisms
3. Some B lymphocytes evolve
into the resting state -
memory cells
 B lymphocytes
and humoral immunity

 Originate from stem cells in bone marrow

 Maturation in bone marrow followed by
migration to secondary lymphoid tissue
 Antigen exposure in secondary lymphoid tissue
 Following exposure to antigen, differentiation
into plasma cells and memory cells
 Plasma cells produce antibodies of all IG classes
ACTIVATION OF ANTIBODY PRODUCING
CELLS BY CLONAL SELECTION

 B lymphocytes recognize intact pathogenic

microorganisms and toxins
 B lymphocytes possess specific surface
receptors for recognition of specific antigen
‒ IgM and IgD
 Binding of specific antigen results in
proliferation of a clonal population of cells
 Antigen determines clonal proliferation
ACTIVATION OF ANTIBODY PRODUCING
CELLS BY CLONAL SELECTION

Proliferation of activated cells is followed

by differentiation into
– Plasma cells
• Life span of
– 4 to 5 days
– 1 to 2 months
• Produce 2,000 antibody molecules / second
– Memory cells
• Life span of years to decades
• Differentiate into plasma cells following stimulation
by same antigen
 PRIMARY AND SECONDARY
ANTIBODY RESPONSE
 Primary Response
– Following exposure to an antigen, there is a slow rise in
IgM followed by a slow rise in IgG

 Secondary Response
– Following exposure to previously encountered antigen,
there is a rapid rise in IgG and slow or no rise in IgM
• Memory or anamnestic response
 Sequential IgM-IgG humoral response

 IgM
– produced as a first response to many antigens (3-5 days)
– levels remain high transiently
 IgG
– produced after IgM (14-17 days)
– higher levels persist in small amounts throughout life
– produced in large amounts during secondary response
• persistence of antigen sensitive ‘memory cells’
after primary response
Dengue infection: immune response
 METHODS IN DIAGNOSTIC
IMMUNOLOGY
Immunochromatographic assay (ICA)
– Antibody or antigen immobilized (Test line)
– Antibody immobilized (Control line)
– Membranes
• Nitrocellulose, cellulose acetate
– Read visually for colored test and control lines
– Examples
• Group A Streptococcus (GAS) antigen
• Influenza A and B antigens
• Respiratory syncytial virus (RSV) antigen
• Rotavirus antigen
• HIV-1/2 antibody
Immunochromatographic assay (ICA)