OUR LADY OF FATIMA UNIVERSITY

MEDICAL LABORATORY SCIENCE BATCH 2024

IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

COURSE OUTLINE: PRELIMS  An English country doctor by the name of Edward Jenner
1. Immunology and Serology was able to successfully prevent infection with smallpox
2. Types of Vaccines by injecting a more harmless substance—cowpox—from
3. Natural vs Adaptive Immunity a disease affecting cows.
4. Phagocytosis  Louis Pasteur: often called the Father of Immunology
5. Acute-Phase Reactants  He observed by chance that older bacterial cultures
6. Inflammation would not cause disease in chickens
7. Active vs Passive  Subsequent injections of more virulent organisms
8. Natural vs Artificial had no effect on the birds that had been previously
9. The Lymphoid System exposed to the older cultures.
10. T and B cells Maturation  He discovered the first attenuated vaccine; this
11. Nature of Antigens event can be considered the birth of immunology.
12. Major Histocompatibility Complex  Attenuation: make a pathogen less virulent; it takes
13. Cytokinesis place through heat, aging, or chemical means.
 Basis for many of the immunizations that are used
REFERENCE BOOKS today.
Stevens, C. D. (2016). Clinical Immunology and Serology A  Pasteur applied this same principle of attenuation to
Laboratory Perspective Fourth Edition. F.A. Davis Company the prevention of rabies in affected individuals

INNATE VERSUS ADAPTIVE IMMUNITY

AN OVERVIEW OF
A. Natural Immunity
IMMUNOLOGY
 It is the individual’s ability to resist infection by
IMMUNOLOGY: means of normally present body functions.
 No prior exposure is required
It can be defined as the study of a host’s reactions when  The response lacks memory and specificity
foreign substances are introduced into the body.
B. Adaptive Immunity
Defined as resistance to disease, specifically infectious
disease.  A type of resistance that is characterized by
specificity for each individual pathogen, or microbial
Study of the molecules, cells, organs, and systems agent, and the ability to remember a prior exposure
responsible for the recognition and disposal of foreign
(nonself) material.
INTRODUCTION TO IMMUNOLOGY
The desirable and undesirable consequences of immune  Study of the reaction of a host when foreign substances
interactions; and the ways in which the immune system can
are introduced into the body
be advantageously manipulated to protect against or treat
disease.  Study of all aspects of body defenses, such as antigens
and antibodies, allergy and hypersensitivity
Antigens - Foreign substances that induce a host response

Antibodies - Serum proteins produced by certain ROLE OF IMMUNE SYSTEM

lymphocytes when exposed to a foreign substance and they
react specifically with that foreign substance. a. Defending the body against infections
b. Recognizing and responding to foreign antigens
Immunity and Immunization - Immunology as a science has
c. Defending the body against the development of tumors
its roots in the study of immunity: the condition of being
resistant to infection. d. The function of the immune system is to recognize self
from non-self and to defend the body against non-self.
- The first recorded attempts to deliberately induce immunity
Such a system is necessary for survival.
date back to the 1500s when the Chinese inhaled powder
made from smallpox scabs in order to produce protection
against this dreaded disease. DEFINITION OF TERMS
 VARIOLATION Antigen
 VACCINATION Antibody
Attenuated
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

Cell of Differentiation (CD) cowpox lesion and then exposed them to smallpox. He
Interleukins (IL) thus proved that immunity to cowpox, a very mild
Major Histocompatibility Complex (MHC) disease, provided protection against smallpox.
Immunity  This procedure of injecting cellular material became
Immunization known as vaccination, from vacca, the Latin word for
Neutrophil ―cow.‖ The phenomenon in which exposure to one agent
Lymphocyte produces protection against another agent is
Monocyte known as cross-immunity. Within 50 years of this
Eosinophil discovery, most of the European countries had initiated a
Basophil compulsory vaccination program.
Dendritic Cell
Complement LOUIS PASTEUR
Tumor Necrosis Factor (TNF)
 In working with the bacteria that caused chicken cholera,
Cytokines
Louis Pasteur, a key figure in the development of both
Chemokinesis
microbiology and immunology, accidentally found that old
Chemotaxis
cultures would not cause disease in chickens.
Complement
 Subsequent injections of more virulent organisms had no
Acute Phase Reactants
effect on the birds that had been previously exposed to
Inflammation
the older cultures. In this manner, the first attenuated
vaccine was discovered. Attenuation, or change, may
HISTORY occur through heat, aging, or chemical means, and it
IMMUNITY AND IMMUNIZATION remains the basis for many of the immunizations that are
 The first written records of immunological used today.
experimentation date back to the 1500s, when the
Chinese developed a practice of inhaling powder made FIRST VACCINATION BY LOUIS PASTEUR
from smallpox scabs in order to produce protection  Pasteur applied this same principle of attenuation to the
against this dreaded disease. This practice of deliberately prevention of rabies, a fatal disease at that time. Although
exposing an individual to material from smallpox lesions he did not know the causative agent, he recognized that
was known as variolation the central nervous system was affected, and thus he
 Variolation (inoculation)- method of scratching the skin studied spinal cords from rabid animals. He noted that
and applying pulverized powder from a smallpox scab spinal cords left to dry for a few
 Variolation -fresh material taken from a skin lesion of a
person recovering from smallpox was subcutaneously days were less infectious to laboratory animals than fresh
injected with a lancet intothe arm or leg of a nonimmune spinal cords.
 In 1885, when a boy who was severely bitten by a rabid
―cow. dog was brought to his home, Pasteur tried out his new
procedure. The boy received a series of 12 injections
Edward Jenner beginning with material from the least infectious cords
 Further refinements did not occur until the late 1700s, and progressing to the fresher, more infectious material.
when an English country doctor by the name of Edward Miraculously, the boy survived, and a modification of this
Jenner discovered a remarkable relationship between procedure is the standard treatment for rabies today.
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
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

CHARACTERISTICS OF CONVENTIONAL
VACCINES

NATURAL VS ADAPTIVE IMMUNITY

Innate, or natural immunity, is the individual’s ability to
resist infection by means of normally present body functions.
These are considered non-adaptive or nonspecific and are
the same for all pathogens or foreign substances to which
one is exposed. No prior exposure is required and the
response lacks memory and specificity. Many of these
mechanisms are subject to influence by such factors as
nutrition, age, fatigue, stress, and genetic determinants
Overview

Acquired immunity, in contrast, 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.

Both systems are essential to maintain good health; in

fact, they operate in concert and are dependent upon
one another for maximal effectiveness.

Natural immunity is responsible for both first and second line

of defense, while adaptive immunity is responsible for the
third line of defense

COMPARISON BETWEEN NATURAL AND

ACQUIRED IMMUNITY
NATURAL/ INNATE IMMUNITY
• Present at birth
• Standardized response for all Antigen
• Lacks memory
• Responsible for the first and second line of defense
in the body
• Pathogen recognized by receptors encoded in the
germline
• Receptors have broad specificity
• Immediate response
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

ADAPTIVE/ ACQUIRED IMMUNITY THE NATURAL IMMUNITY

• Not present at birth Second Line of Defense
• Diverse response for each antigen I.ACUTE PHASE REACTANTS
• Capable of recalling previous antigen thus with II.PHAGOCYTOSIS
memory III. Pathogen-Associated Molecular Patterns (PAMPS) and
• Secondary immune response is greater than Pattern Recognition Receptors (PRR)
primary immune response IV. INFLAMMATION
• Responsible for the Third line of defense in the
body I. Acute Phase Reactants
• Pathogen recognized by receptors generated normal serum constituents that increase rapidly by at least
randomly 25 percent due to infection, injury, or trauma to the tissues
They are produced primarily by hepatocytes (liver
• Receptors have very narrow specificity; i.e., parenchymal cells) within 12 to 24 hours in response to an
recognize a specific epitope increase in certain intercellular signaling polypeptides called
• Slow (3-5 days) response cytokines(e.g IL-6, IL1, and TNF-alpha)
They are indicator of inflammation

THREE LINE OF DEFENSE

1. First Line of Defense (innate immunity) I. ACUTE PHASE REACTANTS
The external defense system which is composed of a. C-Reactive Protein
structural barriers that prevent most infectious agents from CRP is a trace constituent of serum originally thought to be
entering the body. an antibody to the c-polyssacharide of the pneumococci
Elevated levels are found in conditions such as bacterial
COMPOSITION infections, rheumatic fever, viral infections, malignant
 Unbroken skin (pH of 5.5 to 5.6) and the mucosal diseases, tuberculosis and after a heart attack
membrane surfaces, Lactic acid in sweat, and the acidity In addition, CRP binds to specific receptors found on
in GIT and Vagina(pH about 5) , Cilia lining in respiratory monocytes, macrophages, and neutrophils, which promotes
tract, The flushing action of urine phagocytosis.
 Lactic acid in sweat, for instance, and sebum/fatty acids the most widely monitored of the acute-phase reactants
from sebaceous glands maintain the skin at a pH of and is the best indicator of acute inflammation
approximately 5.6. This acid pH keeps most Hs- CRP is a significant risk factor for myocardial infarction
microorganisms from growing. and ischemic stroke
 Lysozyme, an enzyme found in many secretions such as Test: Reverse passive agglutination, Precipitation, RIA,
tears and saliva, and it attacks the cell walls of Complement fixation
microorganisms, especially those that are gram-positive. Serum half-life: 19 hours
 The production of earwax (cerumen) protects the
auditory canals from infectious disease. b. Serum Amyloid A
 Normal microbial flora in the body. Many locations of the It is associated with HDL cholesterol, and it is thought to
body, there is normal flora that often keeps pathogens play a role in metabolism of cholesterol (removing
from establishing themselves in these areas. This cholesterol filled macrophages at the site of tissue injury)
phenomenon is known as competitive exclusion
 Three Line of Defense c. Mannose-Binding Protein
2. Second Line of Defense (innate immunity) a trimer that acts as an opsonin, which is calcium-dependent.
The internal defense system, in which both cells and It is widely distributed on mucosal surfaces throughout the
soluble factors play essential parts. The internal defense body. It has many similarities to the complement component
system is designed to recognize molecules that are unique to C1q, as binding activates the complement cascade and
infectious organisms. helps to promote phagocytosis
COMPOSITION: Lack of MBP has been associated with recurrent yeast
 Phagocytosis, Inflammation, Acute phase reactants, infections
Antimicrobial substances such as complement,
Properdin, Interferon alpha and beta, TNF, and d. Alpha1- Antitrypsin
Betalysin. A general plasma inhibitor of proteases released from
 CELLULAR COMPONENT - Mast cells, Neutrophils, leukocytes, especially elastase.
Macrophages, NK cells It also regulates expression of pro inflammatory cytokines
 HUMORAL COMPONENT- Complement, Lysozyme, such as TNF, Interleukin-1, and IL-6
Interferon Deficiency can lead to premature emphysema and
juvenile cirrhosis
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

e. Ceruloplasmin  Enhancement of phagocytosis by coating of foreign

It is the principal copper-transporting protein in human particles with serum proteins (Ex. Antibodies,
plasma complement c3b, CRP)
It acts as a ferroxidase, oxidizing iron from Fe2+ to Fe3+.  Without the influence of these chemotactic substances,
This may serve as a means of releasing iron from ferritin for cell motion is random (chemokinesis)
binding to transferrin.  Job syndrome / Hyperimmunoglobulinemia E =
A depletion of ceruloplasmin is found in Wilson’s disease, an abnormal chemotactic activity
autosomal recessive genetic disorder characterized by a  Lazy Leukocyte syndrome = abnormal random
massive increase of copper in the tissues (chemokinesis) and chemotactic activity
 Boyden-Chamber Assay (BCA) is the test for
f. Haptoglobin Chemotaxis
Its primary function is to bind irreversibly to free
hemoglobin released by intravascular hemolysis a. Positive chemotaxis –Migration is toward to the site of
Haptoglobin plays an important role in protecting the stimulation
kidney from damage and in preventing the loss of iron by b. Negative chemotaxis–Migration away from the site
urinary excretion. stimulation
However, its most important function may be to provide 4. Engulfment
protection against oxidative damage mediated by free 5. Digestion
hemoglobin, a powerful oxidizing agent that can generate
peroxides and hydroxyl radicals.

g. Fibrinogen
Fibrinogen is the most abundant of the coagulation
factors in plasma, and it forms the fibrin clot
Formation of a clot also creates a barrier that helps prevent
the spread of microorganisms further into the body.
Fibrinogen also serves to promote aggregation of red blood
cells, and increased levels contribute to an increased risk
for developing coronary artery disease, especially in
women

II. Phagocytosis
Elie Metchnikoff, a Russian scientist, observed that foreign
objects introduced into transparent starfish larvae became STEPS INVOLVED IN PHAGOCYTOSIS
surrounded by motile cells that attempted to destroy these (STEVEN’S 3rd Ed.)
invaders, a process called phagocytosis. (A) Adherence: physical contact between the phagocytic cell
Enhancement of phagocytosis by coating of foreign particles and the microorganism occurs, aided by opsonins.
with serum proteins is called a opsonization (B) Outflowing of cytoplasm to surround the
microorganism.
II. PHAGOCYTOSIS (C) Formation of phagosome: microorganism is completely
1. Initiation surrounded by a part of the cell membrane.
Phagocytosis is initiated as a result of tissue damage, either (D) Formation of the phagolysome: cytoplasmic granules
trauma or as a result of microorganism multiplication fuse with membrane of phagosome, emptying contents into
Activated phagocyte has increase surface receptors that this membranebound space.
allow for adherence (E) Digestion of the microorganism by hydrolytic
enzymes
2. Chemotaxis (F) Excretion of contents of phagolysosome to the outside
 Process by which cells tend to move in a certain by exocytosis.
direction under the stimulation of a chemical substances
such as opsonin ( ex. Antibodies, CRP, and complement
components)
 whereby cells are attracted to the site of inflammation by
chemical substances(chemotaxins) such as soluble
bacterial factors or acute-phase reactants including
complement components and CRP
 ‘’Phagos’’– From the greek word ―To prepare for
eating
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

III. Pathogen-Associated Molecular Patterns  _____________ - the process by which cells are
(PAMPS) and Pattern Recognition Receptors (PRR) capable of moving from the circulating blood to the
Three groups of PRRs exist: tissues by squeezing through the wall of a blood vessel
1. Secreted PRRs are molecules that circulate in blood and  Neutrophils are mobilized within 30 to 60 minutes after
lymph; circulating proteins bind to PAMPs on the surface of the injury and their emigration may last 24 to 48 hours
 Migration of macrophages and dendritic cells from
many pathogens. This interaction triggers the complement
cascade, leading to the opsonization of the pathogen and its surrounding tissue occurs several hours later and peaks
speedy phagocytosis at 16 to 48 hours
2. Phagocytosis receptors are cell surface receptors that 3. Resolution and repair- Initiated by fibroblast proliferation,
bind the pathogen, initiating a signal leading to the release of as a result : a. Affected area may be totally repaired b. Injury
effector molecules (e.g., cytokines). Macrophages have cell may lead to the formation of an abscess with some loss of
surface receptors that recognize PAMPs containing function c. Granuloma may be formed, typical of delayed
mannose. hypersensitivity or cell mediated immunity
3. Toll-like receptors (TLRs) are a set of transmembrane
receptors that recognize different types of PAMPs. THE CARDINAL SIGNS OF INFLAMMATION
 TLRs are found on macrophages, dendritic cells, a. Rubor - redness(erythema)
neutrophil and epithelial cells. b. Dolor- pain
 TLR2 recognizes teichoic acid and peptidoglycan c. Calor – Heat
found in gram-positive bacteria - TLR4 recognizes d. Tumor – Swelling(edema)
lipopolysaccharide found in gram-negative bacteria e. Function laesa – loss or diminished function
 TLR1 recognizes lipoprotein found in mycobacteria
 TLR5 recognizes bacterial flagellin  The acute inflammatory response acts to combat the
early stages of infection and also begins a process that
Consequently, the innate immune response may not be able repairs tissue Damage. The predominant WBC found is
to recognize every possible antigen, but may focus on a few neutrophil
large groups of microorganisms, called pathogen-  When the inflammatory process becomes prolonged, it
associated molecular patterns (PAMPs). The receptors of is said to be chronic. The failure to remove
the innate immune system that recognize these PAMPs are microorganisms or injured tissue may result in continued
called pattern recognition receptors (PRRs); e.g., Toll-like tissue damage and loss of function. The Predominant
receptors). Cells found in chronic inflammation are
monocytes/macrophages.
IV. Inflammation
 Inflammation is the body’s response to injury or invasion Third Line of Defense (acquired immunity)
by a pathogen, and it is characterized by increased  If a microorganism overwhelms the body’s natural
blood supply to the affected area, increased capillary resistance, a third line of defensive resistance exists.
 Acquired, or adaptive immunity is a more recently
permeability, migration of neutrophils to the surrounding
tissue, and migration of macrophages to the injured evolved mechanism that allows the body to recognize,
area. remember, and respond to a specific stimulus, an
 Refers the overall reaction of the body to injury or antigen.
 Adaptive immunity can result in the elimination of
invasion by an infectious agent
 Both cellular and humoral mechanisms are involved in microorganisms and recovery from disease and the host
this complex highly orchestrated process often acquires a specific immunologic memory. This
 The primary objective of inflammation is to localize and condition of memory or recall (acquired resistance)
eradicate the irritant and repair the surrounding tissue. allows the host to respond more effectively if reinfection
 Hypoxia can induce inflammation. Inflammation in with the same microorganism occurs.
response to hypoxia is clinically relevant. Ischemia in
organ grafts increases the risk of inflammation and graft Composition:
failure or rejection CELLULAR COMPONENT- T lymphocytes, B
lymphocytes, Plasma cells
HUMORAL COMPONENT- Antibodies, and cytokines
IV. INFLAMMATION
STAGES OF INFLAMMATION
THE ADAPTIVE/ACQUIRED IMMUNITY
1. Vascular response
The key cell involved in the adaptive immune response is the
 Increased blood supply to the infected area (Hyperemia)
Lymphocytes.
 increased capillary permeability caused by retraction of
Adaptive immunity is a type of resistance characterized by:
endothelial cells lining the vessels
• Specificity for each individual pathogen or microbial agent
2. Cellular response
• The ability to remember a prior exposure
 Migration of white blood cells, mainly neutrophils, from
• An increased response to that pathogen upon repeated
the capillaries to the surrounding tissue
exposure
 Migration of macrophages to the injured area
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

Subset of lymphocytes:  a small, flat, bilobed organ found in the thorax, or chest
T cells , B cells, and NK cells (note that NK cells are not cavity, right below the thyroid gland and overlying the heart
involved in adaptive immunity)  Site where T cells mature and develop their identifying
characteristics.
TYPES:  Not fully mature T cells = resides in the cortex
ACTIVE  Mature T cells = resides in the medulla and then released
 Long term immunity
 Slow response 2. Secondary Lymphoid Organs
 The immune system is challenge to produce a A. Spleen
specific antibody  Splenic tissue can be divided into two main types: Red
PASSIVE pulp, White pulp
 Short term immunity  The red pulp makes up more than one half of the total
 Immediate response volume, and its function is to destroy RBCs
 Antibody is readily available  White pulp- contains lymphoid tissue
 Immune system is not challenged, thus, there is no a. PALS (periarteriolar lymphoid sheaths) – Contains mainly
antibody produced by the host T cells
b. Primary follicles – contains naïve B cells
MODE OF ACQUISITION: c. Marginal zone- contains dendritic cells
ACTIVE NATURAL – Infection (chicken pox)
ACTIVE ARTIFICIAL- Vaccine-induced immunity (polio B. Lymph Nodes
vaccine)  are located along lymphatic ducts and serve as central
PASSIVE NATURAL- Transfer in-vivo colostrum collecting points for lymph fluid from adjacent tissues.
PASSIVE ARTIFICIAL- injected antibody (Rabies vaccine  Lymph nodes act as lymphoid filters in the lymphatic
and Snake anti venom) system.
 Lymph nodes respond to antigens introduced distally and
HUMORAL VS. CELLULAR ADAPTIVE IMMUNITY routed to them by afferent lymphatics
Layers
HUMORAL CELLULAR a. Cortex-contains macrophages, follicular dendritic cells,
naïve or resting B cells (in primary follicle), proliferating B
cells and plasma cells (in 2ndary follicles or Germinal
MECHANISM Antibody Cell mediated
centers)
mediated
b. Paracortex- contains mainly T cells c. Medulla- contains T
CELL TYPE B T lymphocytes cells, B cells, plasma cells, and macrophages
lymphocytes
LYMPHOCYTES
MODE OF Antibodies Direct cell-to-cell contact or
ACTION production soluble products secreted by T Lymphocytes
cells / cytokine production The T lymphocytes or T cells populate the following:
PURPOSE Primary Defense against viral and 1. Perifollicular and para cortical regions of the lymph nodes
defense fungal infections, intracellular 2. Medullary cords of the lymph nodes
against organisms, tumor antigens, 3. Peri arteriolar regions of the spleen
bacterial and graft rejection 4. Thoracic duct of the circulatory system
infection
B Lymphocytes
The B lymphocytes or B cells multiply and populate the
THE LYMPHOID SYSTEM following:
1. Follicular and medullary (germinal centers) of the lymph
nodes
LYMPHOID ORGANS
2. Primary follicles and red pulp of the spleen
1. Primary Lymphoid Organs
3. Follicular regions of GALT
A. Bone Marrow
4. Medullary cords of the lymph nodes
 It can be considered the largest tissue of the body, with a
total weight of 1300 to 1500g in adult
 The bone marrow functions as the center for
____________________________.
 Site where most blood cells mature including B cells and
NK cells.

B. Thymus
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

delete the developing T cell by means of apoptosis, or

programmed cell death.

3. Mature T-cells
 Survivors of selection exhibit only one type of marker,
either CD4 or CD8, and they migrate to the medulla
 T cells bearing the CD4 receptor are termed helper, or
inducer cells, while the CD8-positive (CD8) population
consists of cytotoxic or suppressor T Cells.
 CD4 T cells recognize antigen along with MHC class II
protein, while CD8+ T cells interact with antigen and MHC
class I proteins.
 Helper T cells- acts as the orchestrator/ stimulator of the
THE T LYMPHOCYTE effector mechanisms of the immune response such as
 Represents 60 to 80 percent of circulating lymphocytes in Antibody synthesis, macrophage activation, Cytotoxic T
the peripheral blood cell killing, and NK cell activation
 Lymphocyte precursors called thymocytes enter the  Approximately two-thirds of peripheral T cells express CD4
thymus from the bone marrow antigen, while the remaining one-third express CD8
 Most of the circulating T cells express three of the antigen
following Cell of Differentiation (CD) markers:  T helper cells consist of two subsets, termed Th1 and Th2
o CD 2: sheep red blood cell receptor, the classical T cell cells.
surface marker a. Th1 cells produce interferon gamma (IFN-Y) and tumor
o CD 3: part of T-cell antigen-receptor complex necrosis factor beta (TNF-B), which protect cells against
o CD 4: receptor for MHC class II molecule intracellular pathogens.
o CD 8: receptor for MHC class I molecule b. Th1 subset secretes IL-2, interferon-gamma (IFN-γ) and
Trivia: TNF-β, which are responsible for the activation of cytotoxic T
Normal ratio of CD 4+ : CD 8+ cells _____________ lymphocytes and macrophages
In HIV, ratio of CD 4+ : CD 8+ cells _____________ c. Th2 cells produce a variety of interleukins, including IL-4,
AIDS CD 4+ cell count ________________ IL-5, IL-10, and IL-13. The essential role of the Th2 cells is to
Normal CD 4+ count= 500-1300 ul help B cells produce antibody against extracellular
pathogens.
T-CELL DIFFERENTIATION /MATURATION
Thymic stromal cells include epithelial cells, macrophages, Recently, an additional T-cell subpopulation has been
fibroblasts, and dendritic cells, all of which play a role in T described. These are called T regulatory cells (T reg), and
cell development. Interaction with stromal cells under the they possess the CD4 antigen and CD25. These cells
influence of cytokines, especially interleukin-7 (IL-7), is comprise approximately 5 to 10 percent of all CD4-positive T
critical for growth and differentiation cells. T regs play an important role in suppressing the
1. Double-negative thymocytes immune response to self-antigens. They are critical for
2. Double-positive thymocytes prevention of autoimmunity.
3. Mature T-cells
4. Antigen Activation 4. Antigen Activation
5. Activated T-cells  T cells exposed to antigen
 Activated T lymphocytes express receptors for IL-2, just as
1. Double-negative thymocytes activated B cells
 Lack CD 4 and CD 8 markers  CD 25(+) is the receptor for IL-2 5.
 Actively proliferate in the outer cortex under the influence
of IL-7 5. Activated T- Cells
 Rearrangement of the genes that code for the antigen  T lymphoblasts differentiate into functionally active small
receptor known as TCR lymphocytes that produce cytokines / lymphokines.
 Immune response is known as cell mediated immunity
2. Double-positive thymocytes
 Express both CD 4 and CD 8 THE B LYMPHOCYTE
 Positive selection –occurs in the thymic cortex. T cells  Represents 20-35 percent ( 10-20% other books) of
must recognize foreign antigen in association with class I lymphocyte population
or class II MHC molecules and allows only double-positive  Matures at bone marrow
cells with functional TCR receptors to survive.  Precursor cells in antibody production
 Negative selection – occurs in the thymic medulla. There is  B CELLS ARE THE PRECURSOR OF PLASMA CELLS
a Strong reactions with self-peptides send a signal to
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

1. PRO-B Cells (Progenitors) 5. ACTIVATED B CELLS

2. Pre-B CELLS (Precursors)  Antigen-dependent activation of B cells takes place in the
3. IMMATURE B CELLS primary follicles of peripheral lymphoid tissue.
4. MATURE B CELLS  Activated B cells exhibit identifying markers that include
5. ACTIVATED B CELLS CD25, which is found on both activated T and B cells and
6. Plasma cells acts as a receptor for interleukin-2 (IL-2).
 When B cells are activated in this manner, they transform
B CELL MATURATION/DIFFERENTIATION into blasts that will give rise to both plasma cells and so-
1. PRO-B Cells (Progenitors) called memory cells
 The pro-B cell has distinctive markers that include CD19,  Activated B Cells will turn to an Effector B cell/ Plasma Cell
CD45R, CD43, CD24, and c-Kit or Memory Cell
 Intracellular proteins found at this stage are terminal
deoxyribonucleotidetransferase(TdT) and recombination- 6. Plasma cells
activating genes RAG-1 and RAG-2, which code for  Plasma cells are spherical or ellipsoidal cells between 10
enzymes involved in gene rearrangement and 20 μm in size and are characterized by the presence
 C-Kit on the pro-B cell interacts with a cell surface of abundant cytoplasmic immunoglobulin and little to no
molecule called stem cell factor, which is found on stromal surface immunoglobulin
cells.  This represents the most fully differentiated lymphocyte,
 Rearrangement of genes on chromosome 14, which code and its main function is antibody production.
for the heavy-chain part of the antibody molecule  Plasma cells are not normally found in the blood but are
located in germinal centers in the peripheral lymphoid
2. Pre-B CELLS (Precursors) organs
 When synthesis of the heavy chain part of the antibody
molecule occurs, the pre-B stage begins.
 The first heavy chains synthesized are the μ chains, which MEMORY B / T CELLS
belong to the class of immunoglobulins called IgM. These Memory cells are populations of long-lived T or B cells that
μ chains accumulate in the cytoplasm. have been stimulated by antigen. They can make a quick
 Pre-B cells may also express μ chains on the cell surface, response to a previously encountered antigen. Memory B
accompanied by an unusual light chain molecule called a cells carry surface IgG as their antigen receptor; memory T
SURROGATE LIGHT CHAIN. cells express the CD45RO variant of the leukocyte common
antigen and increased levels of cell-adhesion molecules
3. IMMATURE B CELLS (CAMs), chemical mediators involved in inflammatory
 Immature B cells are distinguished by the appearance of processes throughout the body
complete IgM molecules on the cell surface; This indicates
that rearrangement of the genetic sequence coding for
light chains on either chromosome 2 or 22 has taken place
by this time.
 Other surface proteins that appear on the immature B cell
include CD21, CD 40, and major histocompatibility
complex (MHC) class II molecules
 CD 21 acts as a receptor for a breakdown product of the
complement component C3, known as C3d
 CD40 and MHC class II are important for interaction of B
cells with T cells

4. MATURE B CELLS
 In the spleen, immature B cells develop into mature cells
known as either marginal zone B cells or follicular B cells.
 Marginal B cells remain in the spleen in order to respond
quickly to any blood-borne pathogens they may come into
contact with.
 Follicular B cells migrate to lymph nodes and other
secondary organs. Unlike marginal B cells that remain in
the spleen, follicular B cells are constantly recirculating
throughout the secondary lymphoid organs
 In addition to IgM, all mature B cells exhibit IgD, another
class of antibody molecule, on their surface.
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

In order for the killing to occur, one of the following

mechanism should be followed:
1. INHIBITORY SIGNALS -There is a balance between
activating and inhibitory signals that enables NK cells to
distinguish healthy cells from infected or cancerous ones.
The inhibitory signal is based on recognition of MHC class I
protein, which is expressed on all healthy cells. If NK cells
react with MHC class I proteins, then inhibition of natural
killing occurs. Diseased and cancerous cells tend to lose
their ability to produce MHC proteins.
2. Antibody dependent Cell Cytotoxitiy (ADCC) - NK cells
recognize and lyse antibody-coated cells through a process
called antibody-dependent cell cytotoxicity. Binding occurs
through the CD16 receptor for IgG. Any target cell coated
with IgG can be bound and destroyed.

NATURE OF ANTIGENS & MAJOR

HISTOCOMPATIBILITY COMPLEX
MITOGENS
- Substances that stimulate cell division
NATURE OF ANTIGENS
T Cell Mitogens
 Antigens refers to a substance that reacts with antibody or
 Phytohemagglutinin
sensitized T cells but may not be able to evoke an immune
 Concanavalin A
response in the first place
 Pokeweed
 Immunogens refers to a substance that is capable of
inducing an immune response
B Cell Mitogens
 Thus, all immunogens are antigens but not all antigens are
 Lipopolysaccharide
immunogens
 Staphylococcal protein A
 In discussing serological reactions or particular names of
 Pokeweed
substances such as blood groups, the term antigen is still
more commonly used; hence both terms are used in this
THIRD POPULATION LYMPHOCYTES OR chapter.
NATURAL KILLER CELLS
 Represents 5-10(<10%)percent of total Lymphocyte FACTORS INFLUENCING THE IMMUNE RESPONSE
population. Other Books 10 to 15%  Age
 They are under the NATURAL IMMUNITY, thus they are  Overall health
non-specific compared to T and B cells  Dose
 These lymphocytes are generally larger than T cells and  Route of inoculation
B cells at approximately 15 μm in diameter, and they
 Genetic
contain kidney-shaped nuclei with condensed chromatin
and prominent nucleoli.
TRAITS OF IMMUNOGEN
 They have been named natural killer, or NK, cells
I. Molecular size
because they have the ability to mediate cytolytic
reactions and kill target cells without prior exposure to  MW of at least _________
them.  The greater the size, the more immunogenic
II. Chemical composition & complexity
 THEY POSSESSED CD 16, CD 56, AND CD 94 markers.
 They are essential mediators of virus immunity. Their  Protein = most immunogenic
deficiency in humans lead to uncontrolled viral replication  Carbohydrates= 2nd most immunogenic (less
and poor clinical outcome immunogenic than protein)
 They can destroy virally infected cells or tumor cells  Lipids & Nucleic acid = least or non-immunogenic alone
without/No MHC restriction (MHC-unrestricted cytolysis), III. Foreignness
or antigen presentation.  Able to distinguish self and non-self. Those substance
 NK CELLS, similar with cytotoxic T cells, recognized as non-self are immunogenic.
KILL/ANNIHILATE Virally infected cells or cancerous  The more distant taxonomically the source of the
cells through the secretion of granzymes and perforin. immunogen is from the host, the more successful it is as
a stimulus
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

Autoantigen - Antigen of the same individual MAJOR HISTOCOMPATIBILITY COMPLEX

Alloantigen - Antigen from different individual but of the  Originally referred to as human leukocyte antigens (HLA)
same spp.  Genes coding for the MHC molecules in humans are
Heteroantigen - Antigen from two different spp. found on the short arm of chromosome 6
Heterophile antigens - Heteroantigens that exist in
 Major function is to bring or present antigens in the body
unrelated plants or animals but are either identical or closely
to the surface cells for recognition of T cells.
related in structure so that antibody to one will cross-react
 Regulate immune response and play a role in
with antigen of the other
graft/transplant rejection
IV. The ability to be processed and presented with MHC
 MHC molecules are produced/synthesized in the rough
molecules
endoplasmic reticulum
 Any immunogen that is capable of cross-linking surface
 Transporters associated with antigen processing (TAP1
immunoglobulin molecules is able to trigger B-cell
and TAP2), are responsible for the adenosine
activation.
triphosphate-dependent transport of peptides suitable for
 The immunogen does not necessarily have to be
binding to class I molecules
degraded first. However, for T cells to be able to
 Class I genes are found at three different locations or loci,
recognize an immunogen it must first be degraded into
termed A, B, and C.
small peptides by an antigen-presenting cell (APC). Then
 Class II genes are situated in the D region, and there are
the peptides form a complex with MHC proteins.
several different loci, known as DR, DQ, and DP
 Class III molecules are secreted proteins that have an
NATURE OF EPITOPES immune function, but they are not expressed on cell
 This key portion of the immunogen is known as the surfaces, as are class I and II. Class III molecules also
determinant site or epitope. have a completely different structure compared with the
 Epitopes can be sequential / linear or they can be other two classes. They do not participate in antigen
conformational epitope presentation
 It is the part of antigen/immunogen than binds to the
paratope of the antibody MHC CLASS I
 Paratope = part of antibody that binds to the antigenic 1. Endogenous antigen within cytosol is degraded by
determinant site proteasome.
a. Linear epitopes consist of sequential amino acids on a 2. Peptides transported into endoplasmic reticulum by TAP.
single polypeptide chain. 3. Alpha chain of class I MHC binds Beta macroglobulin
b. Conformational epitopes result from the folding of a 4. Alpha chain of class I MHC binds peptide.
polypeptide chain or chains, and non-sequential amino acids 5. Peptide-class I MHC transported to Golgi complex and
are brought into close proximity. then to cell surface.
6. Class I MHC peptide binds to CD8+ T cell.
HAPTENS AND ADJUVANTS MHC CLASS II
HAPTENS 1. Class II MHC binds invariant chain to block binding of
 Substances that are too small to be recognized by endogenous antigen.
themselves, but if they are complexed to larger 2. MHC complex goes through Golgi complex.
molecules, they are then able to stimulate an immune 3. Invariant chain is degraded, leaving CLIP (class II
response. invariant chain peptide) fragment.
 Haptens are non immunogenic materials that, when 4. Exogenous antigen taken in and degraded and routed to
combined with a carrier, create new antigenic intracellular vesicle.
determinants. Thus, by themselves haptens are antigens 5. CLIP fragment exchanged for antigenic peptide.
but not immunogens 6. Class II MHC antigenic peptide is transported to cell
 Common examples are drugs and therapeutic hormones surface.
7. Class II MHC peptide complex binds to CD4+ T cell.
ADJUVANTS
ADDITIONAL INFORMATION
 An adjuvant is a substance administered with an
immunogen that increases the immune response. IMPORTANCE OF HLA TYPING
 Addition of an adjuvant to a substance used for an 1. Tissue/Organ transplant -must be ABO and HLA matched
immunization helps to make the immunization more a. siblings- 50% chance HLA matched
effective b.parents-25% chance HLA matched
2. Disease associated
 Adjuvants can enhance cell mediated immunity, humoral
immunity, and the macrophage phagocytic system a. HLA B-27= associated with ankylosing spondylitis
―bamboo spine
 Examples are Aluminum salts, and Freund’s complete
b. HLA DR3= associated with SLE
adjuvant (mineral oil, emulsifier, and killed
c. HLA DR4= associated with RA
mycobacterium).
3. Paternity testing – Using HLA-A and HLA-B related
 OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024
IMMUNOLOGY AND SEROLOGY
MS. CAROL JANE LORENZO, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON

METHODS FOR HLA DETECTION The major cytokine families include tumor necrosis factors
1. Serological method (TNF), interferons (IFN), chemokines, transforming
2. Cellular method growth factors (TGF), and colony stimulating factors
3. Molecular method – The preferred test for HLA antigens (CSF). There are also the interleukins (IL), which currently
number from IL-1 to IL-32.

CYTOKINES Cytokines of the Innate immune response

Chemokines, Interferon type 1 (IFN alpha and beta), IL-1, IL-
Cytokines 6, IL-10, 1L-12, IL-15, IL-18, TNF
 These are small soluble proteins that regulate the
immune system, orchestrating both innate immunity and Cytokines of the Adaptive immune response
the adaptive response to infection. Interferon Gamma, IL-2, IL-4,IL-5, IL-13, Lymphotoxin, TGF-
 These chemical messengers, produced by several Beta
different types of cells, have activity-modulating effects on
the hematopoietic and immune systems through Major inflammatory cytokines
activation of cell-bound receptor proteins. TNF-a, IL-1, IL-6, IFN-Gamma
 Cytokines are induced in response to specific stimuli—
such as bacterial lipopolysaccharides, flagellin, or other Major anti-inflammatory cytokines
bacterial products—through the ligation of cell adhesion TGF-Beta, IL-10, IL-13, IL-35
molecules or through the recognition of foreign antigens
by host lymphocytes. MUST KNOW AND LIST OF SOME IMPORTANT
 Cytokines can be categorized under natural immunity or CYTOKINES
adaptive immunity. IL-1 -IL-1α and IL-1β are pro inflammatory cytokines
 Cytokines that are produced by the T cells are known as produced by monocytes, macrophages, and dendritic cells
Lymphokines early on in the immune response
 Cytokines that are produced by the o IL-1 acts as an endogenous pyrogen (induces fever) in
Monocytes/Macrophages are known as Monokines the acute-phase response through its actions on the
 Cytokines are not produced by B cells. hypothalamus

Effects of Cytokine - The effects of cytokines in vivo include IL-2 -T cell and B cell growth factor
regulation of growth, differentiation, and gene expression by
many different cell types, including leukocytes. These effects IL-3 -Known as multi-colony-stimulating factor. It promotes
are achieved through both autocrine stimulation (i.e., hematopoiesis . It stimulates Myeloid, lymphoid, and
affecting the same cell that secreted it) and paracrine (i.e., erythroid lineage
affecting a target cell in close proximity) activities. IL-6 - It is a major factor for production of Acute phase
Occasionally, cytokines will also exert endocrine (i.e., reactants Can be used as a Renal stone marker.
systemic) activities.
TNF-a - Known as cachectin ,produced mainly by the
Pleiotropy - A single cytokine that can have many different macrophages and NK cells Lipopolysaccharide found in
actions. gram negative bacteria – major stimulus for TNF-a
production Secretion of higher levels will lead to septic
Redundancy - Different cytokines activate some of the same shock.
pathways and genes. This redundancy may be explained by
the fact that many cytokines share receptor subunits. For TNF-B - Known as lymphotoxin, produced by T cells For
instance, IL-6, IL-11, leukemia inhibitory factor, oncostatin M, killing and endothelia activation
ciliary neurotrophic factor, and cardio trophin all utilize the
gp130 subunit as part of their receptors. Type 1 IFN
o IFN-alpha (leukocyte interferon) – secreted by
Synergistic reaction - Cytokines often act in networks; if the leukocytes
effects complement and enhance each other. o IFN-beta (Fibroblast IFN /Epithelial cell IFN) –
Antagonism- A cytokine that may counteract the action of secreted by fibroblasts
another cytokine Function: They both inhibits viral replication

Cytokine Storm - A massive overproduction and Type 2 IFN

dysregulation of cytokines produced by hyper stimulation of o Interferon gamma (Immune interferon) – secreted by
the immune response or hyper cytokinemia. Cytokine storms T cells and NK cells
may lead to shock, multi organ failure, or even death, thus Function: Major macrophage activator, and increases
contributing to pathogenesis expression of MHC class 1 and 2