IMMUNOLOGY & SEROLOGY

MITCH ROY BALBACAL MARISTELA, RMT, MD

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This review booklet belongs to:
___________________________________________________________
IMMUNOLOGY & SEROLOGY (50%)
Historical Background
Natural/ Innate Immunity
Acquired Immunity - Humoral
Acquired Immunity - Cell-mediated
Complement System
MHC, HLA, and Transplantation
Immunologic tests (Principles and Procedures)
Tumor Immunology
Hypersensitivity
Instrumentation and Quality Management
Definition of terms in Immunology
1. Immunology - study of the molecules, cells, organs, and systems responsible for the recognition and disposal of
foreign (non-self ) material; how body components respond and interact; the desirable and undesirable
consequences of immune interactions; and the ways in which the immune system can be advantageously
manipulated to protect against or treat disease
a. Study of host immune system and its responses to microbial pathogens and damaged tissues
2. Immune system - collection of cells, tissues, and molecules that mediate resistance to infection
3. Immune response - coordinated reaction of the immune system and molecules to infectious microbes
4. Immunity - state of resistance to disease
5. Antigen (Ag) - any molecule that can bind to the components of the specific immune response
6. Antibody (Ab) - family of defensive proteins the body makes when stimulated by an antigen
a. When secreted by plasma cells, they are termed immunoglobulins
7. Anamnestic/ Secondary response - pertains to the immunologic memory
a. Rapid rise in the immune reaction after re-exposure to an antigen
8. Cytokines - substances, mostly proteins, secreted by cells affecting the behavior of nearby cells
a. Those cells with cytokine receptors
IMMUNITY
 Role of Immune System

1. Defending the body against infections

2. Recognizing and responding to foreign antigens
3. Defending the body against the development of tumors
4. The immune system can injure cells and induce pathologic inflammation
 Protective or Destructive?

Protective yet may be destructive

 1500s

The first written records of immunological experimentation date back to the 1500s, when the
Chinese developed a practice of inhaling powder made from smallpox scabs in order to produce
protection against this dreaded disease. This practice of deliberately exposing an individual to
material from smallpox lesions was known as variolation. The theory was that if a healthy
individual was exposed as a child or young adult, the effects of the disease would be minimized.
However, this was not always the case.
 1700s - Edward Jenner

Edward Jenner discovered a remarkable relationship between exposure

to cowpox and immunity to smallpox. After observing the fact that milkmaids who were
exposed to cowpox had apparent immunity to smallpox, he deliberately injected
individuals with material from a cowpox lesion and then exposed them to smallpox. He thus
proved that immunity to cowpox, a very mild disease, provided protection against smallpox. This
procedure of injecting cellular material became known as vaccination, from vacca, the Latin
word for “cow.”

The phenomenon in which exposure to one agent provides protection against another agent is
known as cross-immunity.
 Louis Pasteur

In working with the bacteria that caused chicken cholera, Louis Pasteur, a key figure in the
development of both microbiology and immunology, accidentally found that old
cultures would not cause disease in chickens. 2 Subsequent injections of more virulent organisms
had no effect on the birds that had been previously exposed to the older cultures. In this
manner, the first attenuated vaccine was discovered.

Attenuation, or change, may occur through heat, aging, or chemical means, and it remains the
basis for many of the immunizations that are used today.

He also employed this principle to rabies.

 First line of Defense
1. Unbroken skin
a. Keratinization
b. Constant epithelial cell renewal - labile
cells
c. Normal microbiota - microbial
antagonism

1. Intact mucosa
a. Mucosal secretions - sebum, nasal
mucus, cerumen, lactic acid in sweat,
stomach acid, intestinal secretions,
acidity of the vagina, saliva, tears
i. Lysozyme - attacks and destroy the
cell wall particularly the
_____________ of gram positive
organisms
ii. Immunoglobulin A

1. Ciliary function
 Natural and Acquired Immunity

Natural, or innate or native, immunity is the ability of the individual to resist infection by
means of normally present body functions.

Acquired, or adaptive or specific, immunity is a type of resistance that is characterized by

specificity for each individual pathogen, or microbial agent, and the ability to remember a prior
exposure, which results in an increased response upon repeated exposure.
 Second and Third line of
Natural Immunity Defense Adaptive Immunity
A. Innate resistance A. Acquired resistance
B. Non-specific B. Clonally distributed receptors
C. Normal body functions C. Specific
D. No memory D. Memory/ Recall
E. N response against antigens E. Diverse response
F. Acute Inflammation F. Uses the cells and molecules of
innate system to eliminate

Key players Key players

 Second and Third line of
Natural Immunity Defense Adaptive Immunity
Phagocytes
- Neutrophils - bacterial and acute
inflammation
- Monocytes
- Eosinophils
- Basophils/ Mast cells
- Lymphocytes - NK cells

Macrophages
1. Connective tissue - histiocytes
2. Lungs - alveolar, dust cells
3. Brain - microglia
4. Kidney - mesangial cells
5. Liver - Kupffer cells
6. Placenta - Hoffbauer cells
 NATURAL
IMMUNITY
Acute Phase
Reactants
Cytokines are mainly produced by

Monocytes and
Macrophages
 Lack of MBP

Recurrent yeast
infections
 Dendritic Cells
Named because they are covered with long membranous extensions that make them resemble nerve cell dendrites.

Their main function is to phagocytose antigen and present it to helper T lymphocytes. While their actual
developmental lineage is not known, they are believed to be descendents of the myeloid line. They are classified
according to their tissue location, in a similar manner to macrophages.

- Langerhans cells are found on skin and mucous membranes;

- Interstitial dendritic cells populate the major organs such as the heart, lungs, liver, kidney, and the
gastrointestinal tract;
- and interdigitating dendritic cells are present in the T lymphocyte areas of secondary lymphoid tissue
and the thymus.

They are the most potent phagocytic cell in the tissue.

 Toll-like Receptors

- Toll is a protein originally discovered in the fruit fly

Drosophila, where it plays an important role in antifungal
immunity in the adult fly
- highest concentration of these receptors occurs on
monocytes, macrophages, and neutrophils
- TLR2 recognizes teichoic acid and peptidoglycan found in
gram-positive bacteria, while TLR4 recognizes
lipopolysaccharide found in gram-negative bacteria
 E. Metchnikoff

Cellular theory of immunity through phagocytosis

Phagocytosis

Steps in Opsonization
Complement component which is
a chemotaxin

C5a
 The principal factor in determining
whether phagocytosis
can occur
physical nature of the surface
of the bacteria
and phagocytic cell
hydrophobic>hydrophilic
 Neutrophil Extracellular Traps
NETs

NETs function in innate immunity. They are composed of chromatin components, including histones, and neutrophil
antimicrobial proteins. Microbes are trapped in NETs, where they encounter high concentrations of antimicrobial
proteins.
 Bacteremia → Sepsis

If not contained by the immune system

Mononuclear phagocyte system
 Mononuclear phagocyte system
Common origin, similar morphology, and shared
functions, including rapid phagocytosis
mediated by receptors for IgG and the major
fragment of C3.
- Fixed
- Wandering
 Mononuclear phagocyte system
Macrophage

- Interferon-gamma (IFN-γ) and granulocyte colony-stimulating factor (G-CSF) activates

macrophage
- Macrophage secretes tumor necrosis factor α (TNF-α, cachectin), which can activate macrophages
itself under certain in vitro conditions.
 Mononuclear phagocyte system
Terminal stage of development

- multinucleated giant cell, which characterizes granulomatous inflammatory diseases such as

tuberculosis.

- Both monocytes and macrophages can be shown in the lesions in these diseases before the
formation of giant cells, thought to be precursors of the multinucleated cells.
Mononuclear phagocyte system
Terminal stage of development
 Cardinal Signs Inflammation

1. Rubor - redness
2. Calor - warmth
3. Dolor - pain
4. Tumor - swelling
5. Functio laesa - loss of function
 Acute Inflammation

5 R’s
1. Recognition of the injurious agent
2. Recruitment of leukocytes
3. Removal of the agent
4. Regulation (control) of the response
5. Resolution (and repair)
 Acute Inflammation

3 components
1. Dilation of small vessels - histamine
2. Increased microvascular permeability - endothelial cell
contraction
3. Emigration of leukocytes and their activation
 Recognition

Receptors (TLRs, Fc receptors, etc)

Circulating proteins (complement)
 Recuitment

1. Margination
a. d/t slower blood flow due to
vasodilation
2. Rolling
a. Selectin (receptor) -- Sialyl Lewis X
(ligand)
3. Adhesion
a. Integrins (receptor) -- CAM (ligand)
4. Diapedesis
a. Postcapillary venules
b. CD31/PECAM-1
5. Chemotaxis
a. Bacterial products with
 Removal of agent
Regulation (control) of response
 Removal of agent
Regulation (control) of response
 Removal of agent
Regulation (control) of response
 Removal of agent
Regulation of response
 Removal of agent
Regulation of response
 Removal of agent
Regulation of response
 Removal of agent
Regulation of response
Interferon (IFN) Gamma - immune interferon

TNF alpha - cachectin; from macrophages

TNF beta - lymphotoxin; from T cells
Resolution/ Repair
Neutrophil Abnormalities
 DISEASE STATES INVOLVING
LEUKOCYTE
INTEGRINS
Leukocyte adhesion deficiency
- history of delayed separation of the umbilical cord, gingivitis, recurrent
and persistent bacterial or fungal skin infections, and impaired wound
healing. A lack of pus formation has also been noted.
M-M Abnormalities
COMPLEMENT
SYSTEM
 Complement Activation
Pathways

CLASSIC ALTERNATIVE LECTIN

Antigen-antibody complex Microbe surface molecules Mannose or other sugars on

IgG or IgM mediated microbial surface
Complement
Complement
 Opsonization

C3b, IgG
 Anaphylatoxins

C3a, C4a, C5a

Membrane Attack Complex

C5b-9
 Complement regulatory proteins

DAF/ CD55
MIRL/ CD59
C1 esterase inhibitor
 Complement Deficiencies
Early complement Increased risk of severe, recurrent pyogenic sinus and respiratory tract infections.
deficiencies (C1-C4) Increased risk of SLE.

Terminal complement Increased susceptibility to recurrent Neisseria bacteremia.

deficiencies (C5–C9)

PIGA gene defect Paroxysmal nocturnal hemoglobinuria

preventing the formation Complement-mediated intravascular hemolysis → haptoglobin, dark urine
of GPI anchors for
complement inhibitors

C1 esterase inhibitor Hereditary angioedema

deficiency Unregulated activation of kallikrein → bradykinin → inflammation
Decreased C4 levels
Complement Deficiencies
Complement Deficiencies
 NK cells
(Third Population)

Inhibitory receptors - MHC recognition

- KIRs (killer cell immunoglobulin-like receptors)
- ILT/LIR
- and CD94/NKG2A receptors

Activatory signals → perforins and granzymes

- MICA and MICB
- CD94/NKG2C and CD94/NKG2D
 NK cells
(Third Population)

- Perforins are pore-forming proteins that polymerize in the presence of Ca2  and
form channels in the target cell membrane.
- Granzymes are packets of serine esterase enzymes that may enter through the
channels and mediate cell lysis.
Antibody-dependent
Cell Cytotoxicity
ADAPTIVE
IMMUNITY
 Lymphoid Structures

Primary/ Central/ Generative

- Bone Marrow
- Thymus
Where B and T cells become mature and become
competent to respond to antigens

Secondary/ Peripheral
- Spleen, Lymph nodes, Tonsils, Peyer’s
patches
Where adaptive immune responses are initiated
 Lymphoid Structures

- The spleen is the largest secondary lymphoid

organ, having a length of approximately 12
cm and weighing 150 g in the adult. It is
located in the upper-left quadrant of the
abdomen, just below the diaphragm and
surrounded by a thin connective tissue
capsule. The organ can be characterized as a
large discriminating filter, as it removes old
and damaged cells and foreign antigens from
the blood.
- Periarteriolar lymphoid Sheath (PALS)
 Lymphoid Structures
- Lymph nodes are located along lymphatic ducts and serve as central collecting points
for lymph fluid from adjacent tissues. Lymph fluid arises from passage of fluids and low
molecular-weight solutes out of blood vessel walls and into the interstitial spaces
between cells.
- Enter via afferent vessels and HEV
- Cortex - B cells
- Paracortex - T cells
 Lymphoid Structures
MALT - GI, GU, respiratory
Tonsils
Appendix
The epidermis contains a number of intraepidermal lymphocytes.
Most of these are T cells, which are uniquely positioned to combat any antigens that enter
through the skin. This association of lymphocytes is known as the cutaneous-associated
lymphoid tissue.
Lymphocytes
T and B cells
Membrane glycoprotein found in
all hematopoietic cells

CD45
B cell Differentiation
B cell Differentiation
Positive and Negative
Selection
Adaptive Immunity
 T-cell and B-cell Activation
T-cell Activation
Mitogen - peptide that induces cell division

T cells B cells
1. Phytohemaglutinnin 1. LPS
2. Concanavalin A 2. Poke Weed Mitogen
3. Poke Weed Mitogen
 Definition of terms

ANTIGEN - substance that reacts with antibody or sensitized T cells but may not be able
to evoke an immune response in the first place.

IMMUNOGEN - macromolecules capable of triggering an adaptive immune response by

inducing the formation of antibodies or sensitized T cells in an immunocompetent host.

ALL IMMUNOGENS are antigens, but NOT ALL ANTIGENS are immunogens.
 Definition of terms

EPITOPE/ DETERMINANT SITE - key portion of the immunogen, molecular shapes or

configurations that are recognized by B or T cells capable of triggering specific
antibody production or a T-cell response

PARATOPE - antigen-binding site

HAPTEN - non-immunogenic non-protein materials that, when combined with a

carrier, create new antigenic determinants; low MW

CARRIER MOLECULE - protein part and is immunogenic; high MW

Definition of terms
 Definition of terms

AUTOANTIGENS are those antigens that belong to the host.

ALLOANTIGENS Are from other members of the host’s species, and

these are capable of eliciting an immune response.
They are important to consider in tissue transplantation
and in blood transfusions.

HETEROANTIGENS are from other species, such as other animals, plants,

or microorganisms

● HETEROPHILE are heteroantigens that exist in

unrelated plants or animals but are either identical or
closely related in structure so that antibody to one will
cross-react
with antigen of the other. Ie. human blood group A and
B antigens, which are related to bacterial
polysaccharides
 Definition of terms

AUTOGRAFT From same individual

ALLOGRAFT From different individual

HETEROGRAFT/ XENOGRAFT From different species

ISOGRAFT From identical twins

 Definition of terms

ADJUVANT - is a substance administered with an immunogen that increases the immune

response. It acts by producing a local inflammatory response that attracts a large
number of immune system cells to the injection site

e.g. Aluminum salts, Freund’s complete adjuvant

Freund’s complete adjuvant

- mineral oil
- Emulsifier
- Killed mycobacteria (0.5 mg/mL)
Antibody Structure
and Function
Antibody Structure
and Function
 Affinity and Avidity

_____________Individual antibody-antigen interaction

_____________Cumulative binding strength of all antibody-antigen interactions in a

multivalent molecule
 Immunoglobulin Isotypes
Structure FUNCTION

Main antibody in secondary response to an antigen

IgG
Most abundant in serum
Crosses the placenta
Can fix, opsonize and neutralize

Produced in the primary (immediate) response to an antigen

IgM
Can fix
Monomer in B cell, Pentamer in secretions

Most produced antibody overall, low in serum; Monomer in circulation,

IgA
dimer in secretions; GI secretions, tears, saliva, mucus, breast milk
Picks up secretory component in epithelial cells → protects the Fc portion
from luminal proteases

Unclear function
IgD
On the surface of many mature naive B cells and in serum

Type I hypersensitivity reaction

IgE
Binds mast cells and basophils and cross-linking
Parasitism
Immunoglobulin Isotypes
 Antigen type and Memory
Antigen type Notes

Antigens lacking a peptide Cannot be presented by MHC to T cells

Thymus-
Weakly immunogenic
independent component Vaccines often require boosters and adjuvants
antigens - lipopolysaccharides - PPSV23 vaccine

Antigens containing a Class switching and immunologic memory occur as a result of direct
Thymus-
protein component contact of B cells with Th cells
dependent - PCV13
antigens
Antigen activation
Laboratory ID of
Lymphocytes
 Cell Flow Cytometry

Automated system for identification of cells

Based on scattering of light
Cells flow in a single file through a beam laser
Forward LS CELL SIZE
Side LS/ 90 deg/ Right Angle CELL COMPLEXITY/ GRANULARITY
Manual ID of
Lymphocytes
Types of Acquired Immunity
 Vaccination
Type Description Comments Examples

Live attenuated Lose pathogenicity Lifelong BCG

Capable of inducing BOTH cellular and C/I in immunosuppressed Rotavirus
humoral response and pregnant MMR & Varicella
Polio (Sabin)
Influenza (intranasal)
Typhoid (Ty21a, oral)
Smallpox
Yellow fever

Killed/ Inactivated Heat/ chemical/ ageing Safer Rabies

Epitope maintained Weaker Influenza
Capable of inducing HUMORAL response Needs booster doses Polio (SalK)
Hepatitis A
Typhoid (Vi polysaccharide,
intramuscular)

Subunit Only the antigens that best stimulate the Low AE Hepatitis B
immune Expensive HPV
system Weaker aP
Hib
PCV13 and PPSV23
Meningococcal

Toxoid Denatured bacterial toxin with Protects vs toxins Tetanus

an intact receptor binding May require boosters as Diphtheria
site antitoxin levels wane
 MHA, HLA and
Transplantation
 Major Histocompatibility Complex
(MHC)
Group of genes encoding for HLA antigens

Genes coding for the MHC molecules in humans are found on the short arm (p arm) of
chromosome 6 at band 21 and are divided into three categories or classes.

Third class are not HLA antigens, but represent complement, TNF, etc.
 MHC
MHC I MHC II
LOCI HLA-A, HLA-B, HLA-C HLA-DP, HLA-DQ, HLA-DR
MHC I loci have 1 letter MHC II loci have 2 letters

BINDING TCR and CD8 TCR and CD4

STRUCTURE 1 long chain, 1 short chain 2 equal-length chains (2 α, 2 β)

EXPRESSION All nucleated cells, APCs, platelets (except RBCs) APCs

FUNCTION Present endogenous antigens (eg, viral or Present exogenous antigens (eg, bacterial
cytosolic proteins) to CD8+ cytotoxic T cells proteins) to CD4+ helper T cells

ANTIGEN Antigen peptides loaded onto MHC I in RER Antigen loaded following release of
LOADING after delivery via TAP (transporter associated invariant chain in an acidified endosome
with antigen processing)

ASSOCIATED β2-microglobulin Invariant chain

PROTEINS

STRUCTURE See next slide.

 MHC
MHC I MHC II

STRUCTURE
MHC
 HLA Subtypes and Associated
Diseases
HLA ASSOCIATED DISEASE/ CONDITION
A3 Hemochromatosis

B8 Addison disease, Myasthenia gravis, Graves

disease

B27 Psoriatic arthritis, Ankylosing spondylitis,

IBD-associated arthritis, Reactive arthritis

C Psoriasis

DQ2/DQ8 Celiac disease

DR2 Multiple sclerosis, hay fever, SLE,

Goodpasture syndrome

DR3 DM type 1, SLE, Graves disease, Hashimoto

thyroiditis, Addison disease

DR4 Rheumatoid arthritis, DM type 1, Addison

disease

DR5 Hashimoto thyroiditis

 HLA on RBCs

Blood group HLA type

Bennett-Goodspeed

Bga HLA-B7

Bgb HLA-B17

Bgc HLA-A28
 Immunologic tolerance

- Lack of response to antigens upon exposure of lymphocytes to antigens. The

lymphocytes are functionally inactivated or killed resulting in tolerance.
- May be induced during encounter of self-antigen with developing lymphocytes in
the primary or secondary lymphoid tissues.

Immunogenic response - activation, proliferation and differentiation of lymphocytes

into effector cells
Tolerogenic response - lymphocytes are either killed or inactivated
Immunologic ignorance - antigen-specific lymphocytes will not produce any reactions
 Immunologic tolerance

Central T cell tolerance Cell death (negative selection) and

generation of regulatory T cells

Peripheral T cell tolerance Functional inactivation of T cells (anergy) or

death (activation induced cell death) or
suppression by regulatory T cells

Central B cell tolerance Changes in receptor specificity (receptor

editing) or killing (negative selection)

Peripheral B cell tolerance Anergy

 Autoimmunity

Inappropriate immune attack against the body’s own proteins or antigens, mediated by
autoantibodies and self-reactive T cells. It is characterized by breakdown of immune
tolerance.

Principal factors for development:

- Inheritance of susceptibility genes
- Environmental factors
Autoimmune diseases
Autoimmune diseases
Autoimmune diseases
Autoimmune diseases
Autoimmune diseases
Autoimmune diseases
Autoimmune diseases
 What is the most sensitive
method for crossmatching and antibody
identification
Single cell analysis by Flow Cytometry
 Transplantation
What are the indications?

How long a patient waits for a transplant depends on the

following factors:
- Blood type (some rarer than others)
- Tissue type
- Height and weight of transplant candidate
- Size of donated organ
- Medical urgency
- Time on the waiting list
- Distance between donor’s hospital and potential donor organ
- Number of donors in local area over time
- Transplantation center’s criteria for accepting organ offers
Depending on the type of organ needed, some factors are more
important than others.
Transplantation
Transplantation
GVHD
 most efficient and probably the most
economical method available for the prevention
of posttransfusion GVHD

Blood product irradiation

Graft Rejection
Graft Rejection
 Transplantation

The donor and recipient may be incompatible. HLA matching is the primary
consideration in assessing whether a donor is acceptable for a given patient and
overshadows any other non-HLA factors, including ABO incompatibility
- Improves the chances for a successful transplantation
- Promotes engraftment, the process of donated cells beginning to grow and
produce new blood cells in the host
- Reduces the risk of post-transplantation graft-versus-host disease (GVHD)
 Minimum matching levels

- For adults at least 6 out of 8

- For cord blood units at least 4 out of 6
 Immunologic tests
Principles & Procedures
 Antigen-Antibody Complex

- non covalent, reversible, intermolecular forces.

 Affinity

- strength of the reaction between a

single antigenic determinant and
binding site
- sum of the attractive and repulsive
forces
- the equilibrium constant that
describes the antigen-antibody
reaction
 Avidity

- a measure of the overall strength of

binding of an antigen with many
antigenic determinants and
multivalent antibodies.
- Avidity is influenced by both the
valence of the antibody and the
valence of the antigen.
Law of Mass Action
 Specificity

- ability of an individual antibody combining site to react with only

one antigenic determinant
 Sensitivity

- defined by the notion that antibodies may or may not react to

certain antigens under specific conditions.
 Cross reactivity

- ability of an individual antibody

combining site to react with more
than one antigenic determinant
Zonal reactions - False negative results
 Phases of Antigen - Antibody Interaction

- Primary Phase – initial or physical attachment

- Secondary Phase – visible manifestations
- Tertiary Phase – biologic consequences
Primary phase
Primary phase
Secondary phase
Secondary phase
Secondary phase
 Precipitation

- Antigen: precipitation
- Antibody: precipitin
- Soluble nature converted to an insoluble complex
- Occurs when an Ag-Ab reaction results to the cross-linking of the Ab until it
forms a lattice structure
- Requires at least two antigenic determinants per molecule and a bivalent
antibody
 Precipitation
In Solutions In Gel medium

Four Methods: Semisolid support

1. Tube
2. Capillary Agarose – most common and best medium
3. Slide because of its relative negative charge that
4. Ring/ Interfacial enhances the Ag-Ab reaction
Oudin - simple diffusion
Detection of bacterial Ags in tissues

Identification and typing of bacteria

Flocculation Test for Syphilis

C reactive protein test

Identification of blood stains

 Precipitation
Ouchterlony
Precipitation
Electrophoresis
Precipitation Measurement
 Agglutination

- Antigen:
- Antibody:
- Aggregation of particulate matter due to combination with specific antibody
- Requires at least two antigenic determinants per molecule and a bivalent
antibody

Sensitization and Lattice formation

Agglutination
 Agglutination
1. Direct: uses antigens found naturally on the surface of cells
2. Passive: employs particles that are coated with antigens not normally found on
their surfaces (carriers used: RBC, polystyrene latex, bentonite, charcoal)
3. Reverse passive: antibodies rather than antigens are attached to carrier particle
4. Agglutination Inhibition - Based on competition between particulate and soluble
antigen for limited antibody combining sites, and a lack of agglutination is an
indicator of positive reaction
5. Coagglutination - uses bacteria as the inert particles to which the antibody is
attached
Agglutination
 Neutralization
Antigenic activity is stopped (Neutralized) by its specific antibody

Antibody renders the antigenic microorganisms non-infective

Example:

ASO (toxin neutralization)

Viral neutralization
- In vivo test
- In ovo test
- Pock reduction test
- In tissue culture
- Plaque reduction test
Complement fixation
Complement fixation
 Labelled
Immunoassays

Labeling techniques
1. Enzymes - horseradish peroxidase, ALP
2. Radioactive substances - 131I, 125I (the label of choice), tritriated hydrogen
3. Fluorescence - FITC (Fluorescein isothiocyanate), TRITC (Tetra methyl rhodamine isothiocyanate),
Phycobiliproteins
4. Chemiluminescence- Luminol, Acridium esters, Dioxetane phosphate, Peroxyoxalate, Ruthenium
derivatives
 Labelled
Immunoassays
Labelled Immunoassays
Clinical Applications
 Tumor
Immunology
 Cancer Hallmarks

1. Self-sufficiency in growth signals

2. Insensitivity to growth-inhibitory signals
3. Altered cell metabolism (Warburg effect or
aerobic glycolysis)
4. Evasion of apoptosis
5. Limitless replicative potential (immortality)
6. Sustained angiogenesis
7. Ability to invade and metastasize
8. Ability to evade the host immune response
 Tumor antigens

1. Tumor-specific antigens (TSAs) on

chemically induced tumors
2. Tumor-associated antigens (TAAs) on virally
induced tumors
3. Carcinofetal antigens
4. Spontaneous tumor antigens
Tumor Markers
Tumor Markers
Tumor Markers
 Others
BRAF Melanoma, Non-Hodgkin lymphoma,
Papillary thyroid CA, Hairy cell leukemia

BCR-ABL1 Chronic myeloid leukemia (CML), Acute lymphoblastic leukemia (ALL)

BCL-2 Follicular lymphoma

C-MYC Burkitt lymphoma

N-MYC Neuroblastoma

JAK2 Chronic myeloproliferative disorders

KRAS Pancreatic CA, Colon CA, Lung CA

HER2/neu, ER, PR Breast cancer

RET MEN2A, MEN2B, Papillary thyroid CA, Medullary thyroid CA, Pheochromocytoma

ALK Lung adenocarcinoma

Others
 Marker for neuroendocrine tumors

S-100, NSE,
chromogranin A,
synaptophysin
 Antigen for Multiple Myeloma

Bence-Jones
Protein
 Guardian of the genome

P53
Li-fraumeni syndrome
Most commonly mutated TSG
 Governor of the cell

Rb
Osteosarcoma and Retinoblastoma
Hypersensitivity
Hypersensitivity
Hypersensitivity Type I
Hypersensitivity Type II
Hypersensitivity Type II
Hypersensitivity Type II
Hypersensitivity Type
III
Hypersensitivity Type IV
Hypersensitivity Type IV
Blood transfusion
Reactions
 Immune mediators

1. Type I - IgE
2. Type II - IgM, IgG
3. Type III - IgM, IgG
4. Type IV - T cells
 Classical Complement mediated

Type I and II
RIST vs RAST
 XX
Other
ImmuneDiseases
Autoimmune: organ- specific vs systemic
Immunoproliferative disorders
Bacterial infections
Parasitic infections
Toxoplasma
Fungal infections
Spirochete infections
Syphilis
Syphilis
Syphilis
Viral Hepatitis
HBV
HBV Notes
EBV
EBV
EBV
CMV
 HIV
ELISA
Western blot
Flow cytometry
RT-PCR

1 - 9 subgroups
2- 2 subgroups
 HIV
ELISA
Western blot
Flow cytometry
RT-PCR

1 - 9 subgroups
2- 2 subgroups
 HIV
ELISA
Western blot
Flow cytometry
RT-PCR

1 - 9 subgroups
2- 2 subgroups
 HIV
ELISA
Western blot
Flow cytometry
RT-PCR

1 - 9 subgroups
2- 2 subgroups
Dengue, Malaria, Leptospirosis,
Typhoid Notes
Dengue, Malaria, Leptospirosis,
Typhoid Notes
Immunologists
Immunologists
 Must-Know
Immunologists
 “Good luck, Future RMTs!”

Thank
y ou —KUYA MITCH