Introduction to IMMUNOLOGY

General Concepts
Let's get familiar with them
Read DIR-page 1-18

Where is your immune system?

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Immune System lies in your blood

...and somewhere else

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Blood
the average human has
5 litres of blood
it is a transporting fluid
it carries vital
substances to all parts
of the body

X 500
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Plasma (55%): liquid part of blood.

Plasma transports:Soluble food molecules
Waste products
Hormones
Antibodies
Red blood cells (5-6 millions /ml):
transport oxygen, specialised to do this
Also carry some CO2
White blood cells (5000 /ml)
Platelets (150,000 to 400,000 per
mm3):
if you get cut, platelets produce tiny
fibrin threads these form a web-like
mesh that traps blood cells.These
harden forming a clot, or "scab.
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x 1000

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COMPLETE BLOOD COUNT WITH

DIFFERENTIAL (CBC WITH DIFF)
References Ranges
Erythrocytes (RBC)

4.0 to 5.4

M/uL

Thrombocytes (Platelets)

145 to 400

K/uL

Leukocytes (WBC)

4.8 to 10.8

K/uL

40 to 74

Neutrophils
Band neutrophils

0 to 9

Eosinophils

0 to 6

Basophils

0 to 1

Lymphocytes
Monocytes

15 to 47
0 to 12

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ORIGIN OF CELLS OF THE IMMUNE SYSTEM

Derived from common progenitor cell in bone marrow
Pluripotent hematopoietic stem cell

Progenitor Stem Cells

1. Erythroid lineage
Erythrocytes and Megakaryocytes

2. Myeloid lineage
Monocyte/ macrophage, dendritic cells, PMNs, mast cells

3. Lymphoid lineage
Small and large lymphocytes
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CELLS OF INNATE AND ADAPTIVE IMMUNITY

2. 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 (PMNs)
Nuclei are multilobed (2 to 5)

Granulocytes
Cytoplasmic granules
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Diagram

wisegeek.com
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Real Image

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CELLS OF INNATE AND ADAPTIVE IMMUNITY

2. 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

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The real ones

Crawling Macrophage

Neutrophil and DCs

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CELLS OF INNATE AND ADAPTIVE IMMUNITY

3. Lymphoid Lineage
Large lymphocytes (large granular lymphocytes)
Natural killer (NK) cells (CD16, CD56)
Innate immunity to viruses and other intracellular
pathogens
Participate in antibody-dependent cell-mediated
cytotoxicity (ADCC)

Small lymphocytes
B cells (CD19)
T cells (CD3, CD4 or CD8)
Adaptive immunity

Lymphocytes refers to small lymphocytes

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Now, you know how to discriminate the cells of myeloid lineage

But

How to distinguish the cells of

lymphoid lineage?

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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)

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THE CLUSTER OF DIFFERENTIATION (CD)

CD markers on leukocytes (WBC, all types)
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+

Natural killer cell

CD45+, CD16+, CD56+, CD3-

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The immune system

Immune system
Innate (non-specific) immunity

Adaptive (specific) immunity

Anatomic barriers (Skin,mucous

membranes)

Antigen specificity
Diversity

Physological barriers (temperature,

pH)
Phagocytic Barriers (cells that eat
invaders)

Immunological memory
Self/nonself recognition

Inflammatory barriers (redness,

swelling, heat and pain)

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Types Of Immunity
1. Inborn or innate immunity: It is present at birth; This is our
First Line Of Defense.
2. Acquired or specific: It is not present at birth but becomes
part of our immune system as the lymphoid system
develops.
1970: WHO defined immunity as immune response to antigen (
Foreign body) in form of
1. Humoral ( activation of B-lymhocytes)
2. Cellular (by activation of T-lymphocytes)
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Our immune systems generate an almost

infinite variety of cells and substances
Foreign Recognition

Effector Response
To eliminate or neutralize particle

Memory
Upon 2 exposure produces enhanced response

*Problems of the Immune System:

In some cases, the IR fails to function; at other times, the IR can turn on its host

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Humoral and cellular immunity

(antibody mediated or cellular)

AFC

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Discussion Topics
1. Why do warm-blooded, long-lived animals require
particularly complex immune defense?- p4-DIR
2. Why would removal of Ag lead to the decline in an
immune response?- p14-DIR
3. Lets go to Critical Thinking, p18-DIR. Rewrite our
discussion as a homework.
And many more to explore in the DIR textbook

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THE INNATE IMMUNE RESPONSE

Mediated (initiated) by phagocytes, NK cells and soluble proteins
Phagocytes
Cells specialized in the process of phagocytosis
Macrophages
Reside in tissues and recruit neutrophils

Neutrophils
Enter infected tissues in large numbers

Recognize common molecules of bacterial cell surface using a few

surface receptors

Phagocytosis
Capture, engulfment and breakdown of bacterial pathogen

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Something to understand more about phagocytosis and its followed consequences

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THE INNATE IMMUNE RESPONSE

Inflammatory response enhances phagocytosis through acute phase
proteins
Mannose-binding lectin (MBL)
Binds to bacterial surface with particular spatial arrangement of
mannose or fucose
C-reactive protein (CRP)
Binds to phosphorylcholine on bacterial surface
Complement: More info comes up in the next lecture
Set of proteins which bind to bacterial surface
Inflammatory response
Accumulation of fluid and cells at infection site (swelling, redness, heat
and pain)

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an overview on Inflammation

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THE ADAPTIVE IMMUNE RESPONSE

Creates millions of different B and T cells for specific antibody-mediated
and cell-mediated immunity
Antibody-Mediated Immunity (AMI)
Involves B lymphocytes, plasma cells and antibodies
Humoral immunity
Name derives from antibodies found in body fluids (humors - old
medical term)
Cell-Mediated Immunity (CMI)
Involves T lymphocytes, antigen-presenting cells and MHC (major
histocompatibility complex) molecules
Cellular immunity

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ANTIBODY-MEDIATED (HUMORAL) IMMUNITY

Directed against extracellular microorganisms
and toxins
B-lymphocytes (B cells)
Differentiate into plasma cells which produce antibodies
Function as antigen-presenting cells (APCs)

Classification of Antibodies (Immunoglobulins)

Immunoglobulin M (IgM)
Immunoglobulin G (IgG)
Immunoglobulin A (IgA)
Immunoglobulin D (IgD)
Immunoglobulin E (IgE)
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CELL-MEDIATED IMMUNITY (CMI)

Directed against intracellular microorganisms
Non-phagocytic cells and phagocytic cells
T-lymphocytes (T cells)
Differentiate into effector cells following antigen presentation by
antigen presenting cells (APCs)
Functional types of T cells
Helper (CD4 T cells)
TH1 and TH2 cells
Cytotoxic (CD8 T cells)
Regulatory
CD4 and CD8 Tregs

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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

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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)

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THE NATURE OF ANTIBODIES

Antibodies are glycoproteins
Exist as monomers, dimers or pentamers of basic structure
Basic antibody structure has 4 polypeptide chains
2 identical light chains
2 identical heavy chains

Regions of heavy and light chains

Variable
Constant

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THE NATURE OF ANTIBODIES

Also referred to as
Immune globulins / Immunoglobulins (IG)
Immune serum globulins (ISG)
Gamma globulins

Contemporary immunology
Antibody
Secreted form of IG made by plasma cells

Immunoglobulin
Antigen binding molecules of B cells
(B cell antigen receptors)

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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

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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

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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

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ACTIVATION OF ANTIBODY PROCDUCING

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

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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

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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
(APCs)
Antigen presenting cells (APCs)
Macrophages
Dendritic cells
B lymphocytes

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T LYMPHOCYTES AND CELL-MEDIATED IMMUNITY

Antigen presenting cells (APCs)
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

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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
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DISORDERS OF THE IMMUNE SYSTEM

Hypersensitivity Reactions
Over-reaction of adaptive immune response to
harmless antigens
Four Types of reactions (I- IV)

Autoimmunity
Misdirected adaptive immune response
Results from a loss of self-tolerance
Three Types (II, III, IV) of reactions

Immunodeficiencies
Components of immune system either absent or
defective
Genetic or acquired etiology

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IMMUNOLOGY FOR DIAGNOSIS OF DISEASES

Analytical methods using either antibody or

antigen with an indicator system for detecting
specific
Antibodies
Detected using antigens or antibody

Antigens
Detected using antibody

Indicator systems

Latex particles (colored)

Microspheres (colored) conjugated with antibody
Enzymes conjugated to antibody
Fluorochromes conjugated to antibody
Nitrocellulose membranes fixed with antigen or antibody

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METHODS IN DIAGNOSTIC IMMUNOLOGY

Latex agglutination (LA)
Latex particles (dyed) coated with antigen, antibody or?
Read visually for clumping of latex particles

Staphyloslide (Becton Dickinson)

Identification of Staphylococcus aureus
Staphylococcus aureus produces
Coagulase (bound and free)
Protein A
Blue latex particles coated and not coated with
Fibrinogen
IgG

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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

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PRINCIPLES OF OraQuick RAPID HIV-1/2 ICA

ANTIBODY TEST
Antigens and antibody immobilized onto
nitrocellulose membrane in T and C zones
Test (T) Zone
Synthetic peptides from HIV envelope region

Control (C) Zone

Goat anti-human IgG

Developer solution
Facilitates flow of specimen onto test strip
Rehydrates protein-A gold colorimetric reagent

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IMMUNOLOGY FOR PREVENTION OF DISEASE

Hepatitis B
Pre-exposure prophylaxis
Vaccination with hepatitis B surface antigen (HBsAg)

Post-exposure prophylaxis
Administration of
Hepatitis B Immune Globulin (HBIG) Human
Purified IgG antibody from plasma of donors with high
titer of antibody to the hepatitis B surface antigen (antiHBs)

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IMMUNOLOGY FOR TREATMENT OF DISEASE

Rheumatoid Arthritis
Remicade (Infliximab)
IgG kappa monoclonal antibody against tumor
necrosis factor alpha (TNF-alpha)

Breast Cancer
Herceptin (Trastuzumab)
IgG kappa monoclonal antibody against human
epidermal growth factor receptor 2 (HER2)

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