IMMUNOLOGY & SEROLOGY BS MLS

LECTURE/PPT / PROF. JORDAN M. CALLUENG, RMT, MD 3rd Year

[TRANS] UNIT II: BASIC CONCEPTS OF IMMUNOLOGY

 Epitopes can trigger multiple or different

OUTLINE antibodies with multiple specificities
I Introduction  Antigen also refers to a substance that reacts with antibody
II What is an Antigen?
or sensitized T cells but may not be able to evoke an
III What is an Immunogen?
A How and Why We Respond to Particular Immunogens immune response in the first place
i Factors o Mature B cell and T cell who are competent enough to
B Factors Influencing the Immune Response bind or recognize an antigen can evoke or confer an
i Age immune response
ii Overall Health o Both B and T cell can bind antigen because they have
iii Dose their own receptors
iv Route of Inoculation
v General Capacity
C Traits of Immunogens
WHAT IS AN IMMUNOGEN?
i Macromolecular Size  Macromolecules capable of triggering an adaptive immune
ii Chemical Composition and Molecular Complexity response by inducing the formation of antibodies or
iii Foreignness sensitized T cells in an immunocompetent host
iv Ability to be Processed and Presented with MHC o Hapten: elementary, basic or simplest form of antigen
Molecules o Hapten: cannot trigger immune response on its own; it
IV What is an Epitope?
has to bind with carrier/schlepper molecule in order to
A Configuration of Epitopes
B Epitopes/Antigenic Determinants trigger an immune response
C Epitopes Recognized by B Cells may Differ from Those
Recognized by T Cells in a Given Antigen HOW AND WHY WE RESPOND TO PARTICULAR
V What are Haptens? IMMUNOGEN
A Example of Haptens  Individual differences on how and why we respond to
VI What is an Adjuvant
A Examples
antigens
VII Major Histocompatibility Complex
A Genes Coding for MHC Molecules (HLA Antigens) FACTORS
B Three Class of MHC Molecules  It includes
i Structure and Role of Class I Molecule o The nature of the immunogen itself
ii Structure and Role of Class II Molecule
VIII What is Immunoglobulins?
o Genetic coding of MHC molecules that must combine
A Gerald Edelman with an immunogen before T cells are able to respond
B Rodney Porter o Immunogen processing and presentation
C Alfred Nisinoff  How we respond to particular antigen is unique
D Biological Properties of Immunoglobulin Domains like a fingerprint
IX Immunoglobulin Variants
X Immunoglobulin Classes
 This is why some are allergic to peanuts and some
A Immunoglobulin G (IgG) are not
B Immunoglobulin M (IgM)
C Immunoglobulin A (IgA) FACTORS INFLUENCING THE IMMUNE RESPONSE
D Immunoglobulin D (IgD)  Nonspecific factors
E Immunoglobulin E (IgE)
AGE
 Older individuals are more likely to have a decreased
INTRODUCTION
response to antigenic stimulation and neonates do not fully
 ALL IMMUNOGENS ARE ANTIGENS, BUT THE
respond to immunogens
CONVERSE IS NOT TRUE
o Neonates: less than 28 days old; immune system is not
o Not all antigens are immunogenic totally developed yet

OVERALL HEALTH
WHAT IS AN ANTIGEN?
 Individuals who are malnourished, fatigued, or stressed are
 Any substance that can represent antigenic sites
less likely to mount a successful immune response
(epitopes) to produce corresponding antibodies
o Antigenic site or Antigenic determinant: epitope
o Antibody determinant: paratope DOSE
 Small molecules such as haptens and hormones to  Threshold dose for each individual immunogen - innate
macromolecules such as proteins, glycoproteins, immune response to take care of small amounts of
glycolipids and other natural products pathogens and leave the adaptive response for pathogens
 Antigens should have at least one epitope that are present in large
o Antigen can also express multiple epitopes o Each individual has its own threshold for an
 S. Aureus has multiple epitopes on its cell immunogen

CARIAGA, DELA CRUZ. GALANTO CSU BS MLS 3B 1

 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
ROUTE OF INOCULATION
 Actual amount of immunogen needed to generate an
immune response differs with the route of inoculation
o Intradermal
o Subcutaneous
o Oral
o Intramuscular
 EX: Polio vaccine (SABIN) is an oral vaccine;
other vaccine is injected
 SABIN was more effective in triggering immunity
compared to the injected form
 Route of inoculation made a difference on evoking
an immune response
GENETIC CAPACITY
 Predisposition is linked to the MHC (and to the receptors
generated during T and B lymphocyte development
TRAITS OF IMMUNOGENS
 The ability of an immunogen to stimulate a host response
depends on the following characteristics:
 Characteristics that contributes to immunogenicity of
antigen
o Macromolecular size
o Chemical composition and molecular complexity
o Foreignness
o The ability to be processed and presented with MHC
molecules
MACROMOLECULAR SIZE
 The rule of thumb is that the greater the molecular
weight, the more potent the molecule is as an
immunogen
o Mas malaki: mas potent
 Usually an immunogen must have a molecular weight of at
least 10,000 Daltons to be recognized by the immune
system
 The best immunogens typically have a molecular weight
of over 10,000 daltons
 However, there are exceptions, because a few substances
with a MW of less than 10,000 have been known to induce
an immune response
 SUPERANTIGENS: classified as such because of their
macromolecular size and composition and on manner on
binding with receptor on T cells
o They bind in binding cleft and overlap in areas where
MHC molecules are located - kaya nagkakaroon ng
superantigen effect
 Ex: TSS (toxic shock syndrome) exotoxin
 Streptococcal toxic shock syndrome
 Pag nainfect ka mas malala: like anaphylactic
shock
CHEMICAL COMPOSITION AND MOLECULAR
COMPLEXITY
 Proteins and polysaccharides are the best
immunogens
o Both are the fundamental or basic structural
component of microorganisms
 Proteins are powerful immunogens, because they are made
up of variety of units known as amino acids
o B cells recognize structures that project from the
external surfaces of macromolecules, and the more
complexity or branching there is, the easier it is for B
cells to respond.
CARIAGA, DELA CRUZ. GALANTO
o Proteins have epitopes that also stimulate T cells,
which is essential to generating T-cell help in antibody
production
 In contrast, synthetic polymers such as Nylon or Teflon are
made up of a few simple repeating units with no bending or
folding within the molecule, and these materials are non-
immunogenic.
o For this reason, they are used in making artifical heart
valves, elbow replacements, and other medical
appliances.
 Mga prosthetis; eto yung ginagamit panggawa
o Nylon: fishing line or fabric
o Teflon: pambalot sa gripo (puti)
 Both are synthetic and relatively inert, meaning
they are not immunogenic
 Carbohydrates are somewhat less immunogenic than
protein
o If complexed or combine with other macromolecule of
another type like lipids or proteins, they become more
immunogenic
o EX: blood group antigens (Rh, Lewis or ABO)
 As immunogens, carbohydrates most often occur in the
form of glycolipids or glycoproteins
o A,B, and H blood group antigens are glycolipids
o Rh and Lewis antigens are glycoproteins
 Pure nucleic acids and lipids are not immunogenic by
themselves, although a response can be generated when
they are attached to a suitable carrier molecule
o Autoantibodies to DNA that are formed in systemic
lupus erythematosus (SLE) are actually stimulated by
a DNA-protein complex rather than by DNA itself
 ETIOLOGY: Production of DNA protein complex
that trigger autoantibody production, targeting
your own DNA
 Pure DNA is not immunogenic
 Underlying pathology: DNA is complexed with
protein sa mga SLE patients
 Production of neo antigen (the DNA-protein
complex) that triggered production of
antibodies that kills or destroys self-antigens
FOREIGNNESS
 Immune system is normally able to distinguish between self
and nonself antigen
 Those substances recognized as nonself are immunogenic
o Self-antigen: non immunogenic
o Nonself: immunogenic
 This ability is acquired as lymphocytes mature in the
primary lymphoid organ.
o 8 weeks age of gestation: lymphocytes are already
developing
o Bata palang na re recognize mo na ano ang foreign or
not
o If naencounter mo: magkakaroon ka ng response
o NO SELF TOLERANCE: AUTOIMMUNE DISEASES
 Any lymphocyte capable of reacting with self-antigen is
normally eliminated
 Different types of antigen with respect to foreignness
of an immunogen
o AUTOANTIGENS
 Antigens that belong to the host
 Do not evoke immune response under normal
circumstances
 Unless in autoimmune diseases
 Loss of immune tolerance: autoimmune
o Autoantigens will evoke immune
response
o ALLOANTIGENS
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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
 From other members of the host’s species
 From person to person
 Example in cases of kidney transplant
 Capable of eliciting an immune response
 Important to consider in tissue transplantation and
in blood transfusion
o HETEROANTIGEN
 From other species such as other animals, plants,
or microorganisms
 HETEROPHILE ANTIGENS
 Special type of heteroantigen
 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 or the other
o Cross reactivity happens
o Ex: heterophile antibody during EBV
(Epstein Barr Virus) that causes
Infectious mononucleosis (IM)
o Production of heterophile antigen is
detected and is used as a screening test
for the infection
 Prior exposure to plants or animals
that triggered a primary immune
response
 Infection with EBV - person
produces heterophile antibodies that
cross reacts with antigens used as
reagent, for example in Sheep Red
Blood cells
 Sheep Red Blood Cell: Ag used
in reagent for Paul Bunnell test
 Exposure to sheep red blood cells or
anything similar to it = primary
response = exposure to EBV =
produce heterophile = Paul Bunner
Test = detects the heterophile
antibodies
o Anti-A antibody, which is normally found
in individuals with blood types other than
A (type B and type O), is originally formed
after exposure to pneumocci or other
similar bacteria.
o Naturally occurring anti-B antibody is
formed after exposure to a similar
bacterial cell wall product
 ISOAGGLUTININS: naturally
occurring; triggered due to exposure
to antigens present in plants or
animals
o Cross-reactivity can be helpful for certain
diagnostic purposes
 During the early states of IM, a heterophile antibody is
formed, stimulated by an unknown antigen
o This antibody was found to react with sheep red blood
cells
 Paul-Bunnell screening test for mononucleosis
ABILITY TO BE PROCESSED AND PRESENTED WITH
MHC MOLECULES
 T and B cell recognize antigens in context with MHC
molecules
 APC (dendritic cell or macrophages)
endocytosed/phagocytosed microorganisms and they
digest it inside their organelles
o Part of the digested microorganism is presented on
surface of APC with MHC attached to it, for them to be
recognized by the T cell
 For a substance to elicit an immune response, it must be
CARIAGA, DELA CRUZ. GALANTO
subject to antigen processing, which involves enzymatic
digestion to create small peptides or pieces that can be
complexed to MHC molecules to present to responsive
lymphocytes
 If a macromolecule can’t be degraded and presented with
MHC molecules, then it would be a poor immunogen
 The particular MHC molecules produced also determine
responsiveness to individual antigens. Each individual
inherits the ability to produce a certain limited repertoire of
MHC molecules
WHAT IS AN EPITOPE?
 Antigenic determinant of an antigen
 Molecular shapes of configurations that are recognized by
B or T cells
 For proteins, epitopes recognized by B cells may consist of
as few as 6 to 15 amino acids
 Large molecules may have numerous epitopes, and each
one may be capable of triggering specific antibody
production of a T-cell response
CONFIGURATION OF EPITOPES
 Linear: sequential; the primary structure of proteins or its
amino acid sequence
o One antigen can have many antigenic determinant
 Conformational epitopes
o Higher degree of protein structure
o Secondary, tertiary, quaternary
 Alpha helix
 Beta helix
 Domain
 Epitopes are formed by nonsequential structure of amino
acid
o Isang segment nagdidikit, to form a conformational
epitope
EPITOPES/ANTIGENIC DETERMINANTS
 May be repeating copies, or they may have differing
specificities
 Sequential or linear (amino acids following one another on
a single chain)
 Conformational epitopes results from the folding of one
chain or multiple chains, bringing certian amino acids from
different segments of a linear sequence or sequences into
close proximity with each other so they can be recognized
together
EPITOPES RECOGNIZED BY B CELLS MAY DIFFER
FROM THOSE RECOGNIZED BY T CELLS IN A
GIVEN ANTIGEN
 Antigen with several epitopes, pwedeng simultaenously
recognized by T and B cells
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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
o Surface antibody on B cells may react with both linear
and conformational epitopes present on the surface
of an immunogen, triggering B cell activation
 B cell: its receptor is IgM
 Easily accessible
 Those in the surface of immunogen
 If the immunogen is a protein, B cells may
recognize the primary, secondary, tertiary, or even
quaternary structure
 It can recognize linear, sequential or
conformational epitope
 If an immunogen is a polysaccharide, the branch
points of branced chains may contribute most to
recognition
o T-cell epitopes are linear but may be molecules found
anywhere in the cell, rather than strictly surface
molecules
 Inside or outside the infected cell
 Because of the MHC molecules
 Ex: tumor cells manufacture abnormal
proteins inside the cell
 Because of the MHC Class I molecules, the
cancer antigens are presented in the cell
surface, to be recognzied by the T cell
receptors
 T cells recognize an epitope only as a part of a
complex formed with MHC proteins on the surface
of an antigen-presenting cell. The antigen-
presenting cell must process an immunogen first
and degrade it into small peptides for it to be
recognized by T cells.
 B cells : present on surface
 T cell: found anywhere in the cell; have the help of MHC
molecules
WHAT ARE HAPTENS?
 Antigens but not necessarily immunogenic
 Non-immunogenic materials that, when combined with a
carrier (schlepper molecules), create a new antigenic
determinants
o Immunogen na kasi naka bind na sa malaking carrier
o Increase size, increased immune response =
immunogenic
 Once antibody production is initiated, the hapten is capable
of reaction with antibody even when the hapten is not
complexed to a carrier molecule
o Kahit wala nang carrier = pwede na magreact sa
antibody
 However, precipitation or agglutination reactions will not
occur, because a hapten has a single determinant site and
cannot form the cross-links with more than one antibody
molecule that are necessary for precipitation or
agglutination
o Iisang epitope lang yung hapten
o Hindi matetest presence ng hapten Ag using
precipitation/agglutination based test
CARIAGA, DELA CRUZ. GALANTO
 Cross linking important on biological effect or function of
antibody
o Role of antibody
 Bind to antigen
 Bind to different effector cells
EXAMPLES OF HAPTENS
 Poison ivy (Rhus radicans) contains chemical substances
called catechols, which are haptens.
 Once in contact with the skin, these can couple with tissue
proteins to form the immunogens that give rise to contact
dermatitis
o Delayed type of hypersensitivity
o Contact Dermatitis: T cell mediated
 Haptens coupling with normal proteins in the body to
provoke an immune response occurs with certain drug-
protein conjugates that can result in a life-threatening
allergic response.
o Best known example: Penicillin
 Causing penicillin allergy
o It is a hapten mediated drug-protein conjugate immune
response
o Causes type 1 hypersensitivity
WHAT IS AN ADJUVANT
 Substance administered with an immunogen that increases
the immune response
o Used in vaccination
o Add adjuvant to increase titer or level of antibody
production after vaccination
 A local inflammatory response that attracts a large number
of immune system cells to the injection site occurs
 Prolonging the existence of immunogen in the area,
increasing the effective size of the immunogen, and
increasing the number of macrophages involved in antigen
processing
o Ex: hepatitis B vaccine
 With adjuvant
 Hepa B immunogen stay is prolonged
 Adjuvant attracts macrophages or APC sa site of
injected para maprocess nila
EXAMPLES
 Aluminum salts are the only ones approved for clinical
use in the United States, and these are used to complex
with the immunogen to increase its size and to prevent a
rapid escape from the tissues. It must be injected into the
muscle to work. The hepatitis B vaccination is an example
of using this type of adjuvant (intramuscular)
 Freud’s complete adjuvant, which consists of mineral oil,
emulsifier, and killed mycobacteria (0.5 mg/mL). Antigen is
mixed with adjuvant and then injected. It is released slowly
from the injection site.
o Freud’s adjuvant produces granulomas, or large areas
of scar tissue, and thus is not used in humans
o Masyadong malakas as adjuvant nagkakaroon ng scar
sa injection site
 Why do individuals respond to particular immunogens
differently?
 Why is there a major problem in matching organ donors to
recipients?
 How do we resolve disrupted paternity?
o Answer to all the three
 Because of the MHC genes and MHC molecules
known as Human Leukocyte Antigen (HLA)
MAJOR HISTOCOMPATIBILITY COMPLEX
 Every individual is unique in terms of MHC composition
o Like a fingerprint, no two individuals are the same
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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
 Evidence now indicates that the genetic capability to mount
an immune response is linked to a group of molecules
originally referred to as human leukocyte antigens (HLA)
 First described by discovering an antibody response to
circulating white blood cells (hence, HLA)
 Also known as MHC molecules, because they determine
whether transplanted tissues is histocompatible and thus
accepted or recognized as foreign and rejected
 MHC molecules are actually found on all nucleated cells
in the body, and they play a pivotal role in the development
of both humoral and cellular immunity
 Their main function is to bring antigen to the cell surface
for recognition by T cells, because T cell activation will
occur only when antigen is combined with MHC molecules
o B cell only recognizes immunogen / Ag present on
surface of cell
o T cell can recognize found anywhere in the cell (inside
or outside a cell)
 Involved in transfusion reactions, graft rejection, and
autoimmune diseases
 Genes controlling expression of these molecules are
actually a system of genes known as the major
histocompatibility complex (MHC).
GENES CODING FOR MHC MOLECULES (HLA
ANTIGENS)
 The MHC system is the most polymorphic system found
in humans
o Maraming alleles (alternate forms of genes)
 It is thought that this polymorphism is essential to our
survival, because MHC molecules play a pivotal role in
triggering the immune response to diverse immunogens.
 Genes coding for the MHC molecules in humans are found
on the short arm of chromosome 6 (chromosome 6p)
THREE CLASS OF MHC MOLECULES
 Class I
o Encodes for MHC molecule A,B,C
 Class II
o Encodes for MHC molecule D, can be DP, DQ, DR
 Class III
o Center
o Encodes for production of cytokines and compelement
which are part of the humoral component of immune
system
 The MHC system is polymorphic, because there are so
many possible alleles at each location
 For example, at least 580 different alleles of HLA-A, 921
alleles of HLA-B, and 312 alleles of HLA-C have been
identified at this time
CARIAGA, DELA CRUZ. GALANTO
 Alleles are alternate forms of a gene that code for slightly
different varieties of the same product
 The probability that any two individuals will express the
same MHC molecules is very low. An individual’s MHC type
is about as unique as a fingerprint.
 You have two copies of chromosome 6, and thus there is a
possibility of two different alleles for each gene on the
chromosome, unless that person is homozygous (has the
same alleles) at a given location. These genes are
described as codominant, meaning that all alleles that an
individual inherits code for products that are expressed on
cells
 Since the MHC genes are closely linked, they are inherited
together as a package called a HAPLOTYPE
o Alleles that encodes for MHC class are located in
proximity with one another (magkakalapit), you inherit
such as a haplotype (as a package)
o Disputed paternity: uses MHC molecule typing
 Because you inherit half of the genes from your
mother, the other half from your mother
 Class I molecules are the watchdogs ( primary frontliner) of
viral, tumor, and certain parasitic antigens that are
synthesized within the cell
 Class II molecules stimulate CD4 T cells in the case of
bacterial infections or the presence of other material that is
endocytosed by the cell
STRUCTURE AND ROLE OF CLASS I MOLECULE
 Glycoprotein dimer, made up of two noncovalently linked
polypeptide chains
o Alpha chain
o Beta chain
o Beta 2 microglobulin
 Peptide binding cleft:
o Dito nagbi bind Ag for recognition
o Made up of alpha 2 and alpha 1
 Alpha 3 domain
o React with CD8 on cytotoxic T cell
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 Reason why because liver hepatocytes has

undetected MHC molecules
 Liver organ donor: no longer has HLA typoing
for compatibility matching to give to recipient
o Class I antigen is a glycoprotein dimer, amde up of two
noncovalently linked polypeptide chains.

STRUCTURE AND ROLE OF CLASS II MOLECULES

 Virally infected cells and tumor cells

o Virus needs to invade or infiltrate nucleus or organelle
for it to undergo viral replication
 Antigen processing mechanism inside a nucleated cell
infected by cancer
 Defective protein produced by the cell
o Processed/digested by proteosome
o Broken down into smaller particles  Dimer rin, with two polypeptide chain
 Cancerous cell must bind first to MHC molecule Class I o Alpha chain
 Packaged in rough endoplasmic reticulum o Beta chain
 Transported into golgi complex  Each chain has two domain
 Transported in the surface of infected cell  Alpha 1 and alpha 2 domain
 On the surface of the infected cell: Antigen of interest plus  Beta 1 and beta 2 domain
class I molecule  Peptide binding cleft mas flexible
 Class II molecules present on all APC
o Class I: all nucleated cell
o Class II: all APC
 REMINDERS:
o Found primarily on antigen-presenting cells, which
include B lymphocytes (actiavted), monocytes,
macrophages, and dendritic cells
 Dendritic cell: most potent among the APC
o Consist of two noncovalently bound polypeptide chains

 Naghihintay yung CD8 T cell

 CD8: Marerecognize nya yung target cell by way of its
MHC-Ag complex expressed on cell surface
 Binding of TCR receptor of T cell with CD8 molecule as a
coreceptor on target cell = cytotoxic T cell activated =
release cytokines = lysis of target cell
 REMINDERS:
o Class I MHC molecules are expressed on all nucleated  Ag is extracellular microorganism
cells, altough they differ in the level of expression
 First it is endocytosed
 Highest on lymphocytes and low or undetected on
 Digested to more simpler form
liver hepatocytes, neural cells, muscle cells, and
sperm

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 Invariant chain helps Class II molecules to maitain a
stabilized form, to stabilize naive or Class II molecules that
have not yet bind to Ag na wala pang antigen
 Class II plus invariant chain transport to rER
 To golgi complex: packaged
 Class II inside endosome meets the process antigen
 Binding
 Nagu unbind yung invariant chain
 Complex of Class II molecule and Ag is transported and
expressed on surface of APC
 Nagaantay yung T cell
 Narecognize yung dendritic cell
 Class II molecule: co receptor of T cell is CD4 receptor
 After binding, T cell is activated
 Proliferation of T cell
 T cell will ultimately activate B cell for it to proliferate to
plasma cell to produce an antibodies
WHAT ARE IMMUNOGLOBULINS?
 Immunoglobulins are Y-shaped glycoproteins found in the
serum portion of the blood. They are composed of 82 to 96
percent polypeptide and 2 to 14 percent carbohydrate.
 By mass, immunoglobulins make up about 20% of plasma
proteins in healthy individuals.
o Under Clinical Chemistry, we have albumin and
globulin.
 Antibodies are the effector molecules of humoral (B cell-
mediated) immunity. They are immunoglobulins that react
specifically and bind with the antigens that stimulated their
production.
o Immunoglobulins are the general term for both the
naturally occurring Immunoglobulin, meaning structure
that are present before antigen stimulation.
 In serum electrophoresis, it is that last to settle in
the medium, called Gamma Globulins.
 Note that the fastest migrating protein is Alpha,
Beta…last is Gamma
o Antibodies is a type of Immunoglobulin, which is
produced in response to antigen exposure. This is
more specific compared to Immunoglobulins.
 Antibody activity is associated with the slowest migrating
proteins on electrophoresis: the Y-globulins.
 Lecture Discussion:
o The function of immunoglobulin is supported or actually
due to its structure or anatomy.
CARIAGA, DELA CRUZ. GALANTO
o One immunoglobulin has heavy chain (the long one)
and light chain (shorter one).
o Note that it is like a mirror image, which is identical to
each other. The heavy chain is identical to the left and
same as through in the light chain.
o The light chain has two domains: variable and constant
region of the light chain
o The heavy chain has four domains: variable region of
the heavy chain and three constant regions of the
heavy chain. It can also be four, depending on the type
of immunoglobulin. Later on, you’ll find out that IgM,
which is the biggest, has not only three constant heavy
chain, but more.
o Refer to the upper boxed illustration (coded yellow): it
is the variable region (domain of light chain and heavy
chain – it is where the antigen will bind). The variable
region is also responsible for the specificity of the
antibody towards the antigen.
 Hinge region: responsible on the cross-linking of
antigens.
 Constant Domains of the heavy chains: the site for
complement binding.
 CH3: where macrophages and effectors combine
that will ultimately kill the infected cells.
 It has two identical antigen-binding sites (bivalent) at the
tips of the Y (Fab region) and binding sites for complement
components and/or various cell surface receptors on the tail
of the Y (Fc region).
 Each immunoglobulin molecule is composed of two
identical heavy (H) chains and two identical light (L) chains.
These polypeptide chains are held together by noncovalent
interactions that are stabilized by disulfide bonds.
 Parts of both the H and L chains form the antigen-binding
sites. Each immunoglobulin L and H chain consists of a
variable region of about 110 amino acid residues at its
amino-terminal end (the tips of the Y), which forms the
antigen-binding site, linked to a constant region.
 The H constant region is three or four times larger than that
of the L chain. Each chain is composed of repeating,
similarly folded domains: An L chain has one variable
region (VL) and one constant region (CL) domain, whereas
an H chain has one variable region (VH) and three or four
constant region (CH) domains. The amino acid sequence
variation in the variable regions of both L and H chains is
for the most part confined to three small hypervariable
regions that come together at the amino-terminal end of the
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molecule to form the antigen-binding site. Each antigen- BIOLOGICAL PROPERTIES OF IMMUNOGLOBULIN
binding site is only large enough to bind an antigenic DOMAINS
determinant the size of five or six sugar residues.
DOMAIN KNOWN OR PROBABLE FUNCTION
CH3 1. Cytotoxic reactions involving:
a. Macrophages and monocytes
b. Heterologous mast cells
c. Cytotoxic killer cells (K) cells
d. B cells
2. Noncovalent assembly of heavy and light chains
CH2 1. Binding of complement (C1q)
2. Control of catabolic rate
CH1/CL 1. Noncovalent assembly of heavy and light chains
2. Covalent assembly of heavy and light chains
3. Spacers between interdomain interactions involving
antigen binding and effector functions VH/VL1
1. Antigen binding
2. Noncovalent assembly of heavy and light chains

 Papain Digestion = 2 identical Fab fragments + 1 Fc

fragment
 Pepsin Digestion = 1 F(ab)2 fragment + 1 Fc Fragment
 The protective effect of immunoglobulins is not due simply
to their ability to bind antigen.
 They engage in a variety of biological activities mediated by
Fc region of the molecule.
 The Fc fragment of the antibody molecule determines what
will happen to the antigen once it is bound.
 Immunoglobulins with the same antigen-binding capacity
can have a variety of different Fc regions and therefore
different functional properties, such as activating the
complement system and attaching to Fc receptors on
macrophages, thereby aiding phagocytosis of antigens.
o Note that the difference lies on the composition of  Example: CH3, is where the site of macrophages,
monocytes, cytotoxic cells, etc. binds.
heavy chains, especially for Ig. Unlike for light chains
o Another, CH2, is where complements bind (C1q – the
that it is either Kappa or Lambda only.
first complement) binds.
o C1H, where it has biological property, particularly on
GERALD EDELMAN
the stabilization of immunoglobulin.
 Responsible for identifying that immunoglobulin is
composed of a heavy chain and a light chain.
IMMUNOGLOBULIN VARIANTS
 He isolated / measured the fragments, and was later called
heavy chain and light chain.
RODNEY PORTER
 Use Papain Enzyme
 If immunoglobulin is digested with Papain, the cleavage will
form on the upper part of disulfide bond, resulting to two
identical Fab fragments.
 End result: two monovalent bond and an intact FC.

ALFRED NISINOFF
 Used Pepsin Enzyme to further determine the structure of
Immunoglobulin.
 He found out that when you digest your Ig with pepsin, you
produce a 1 Fab, 2 FC Fragments
 Where is Pepsin found: Stomach, which is responsible for
protein digestion.
 End result: A cleavage of pepsin is right below the disulfide  Variants: dictated on the different structures of the Ig. This
bond. However, if disulfide bond is lost, FC fragments will happens because of gene rearrangement on the genes that
be lost / scattered. code for immunoglobulin.
o Isotype: the same heavy chain for each class.
o Allotype: variations in constant regions.

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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY

o Idiotype: variations in variable region.

Table shows the different variations in immunoglobulin

.
IMMUNOGLOBULIN CLASSES

 GMADE: Order of particular immunoglobulin class in the  IgG3 has the largest hinge region and the largest number
serum of interchain disulfide bonds; therefore, it is the most
o IgG – has the highest level of concentration in the efficient at binding complement, followed by IgG1.
serum. Followed by IgM, IgA, IgD, IgE.  Major functions of IgG include the following: (1) providing
 The important thing to know in the table above is its Valency immunity for the newborn because IgG can cross the
for antigen binding placenta; (2) fixing complement; (3) coating antigen for
enhanced phagocytosis (opsonization); (4) neutralizing
IMMUNOGLOBULIN G (IGG) toxins and viruses; and (5) participating in agglutination and
 The predominant immunoglobulin in humans, precipitation reactions.
approximately 75 to 80 percent of the total serum o Opsonization is the process by which an opsonin
immunoglobulins. surrounds an antigen on binds to an antigen for it to be
 has the longest half-life of any immunoglobulin class, recognized by a phagocyte, for enhanced
approximately 23 to 25 days, which may help to account for phagocytosis.
its predominance in serum.  All subclasses of IgG appear to be able to cross the
 four major subclasses: IgG1, 67 %; IgG2, 22 %; IgG3,7 %; placenta, although IgG2 is the least efficient.
and IgG4,4 %.  Macrophages, monocytes, and neutrophils have receptors
 Variability in the hinge region affects the ability to reach for on their surfaces that are specific for the FC region of IgG.
antigen and the ability to initiate important biological This enhances contact between antigen and phagocytic
functions such as complement activation. cells and generally increases the efficiency of phagocytosis.

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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
 IgG1 and IgG3 are particularly good at initiating
phagocytosis, because they bind most strongly to FC
receptors.
 IgG has a high diffusion coefficient that allows it to enter
extravascular spaces more readily than other
immunoglobulin types. In fact, it is distributed almost
equally between the intravascular and extravascular
spaces. Thus, it plays a major role in neutralizing toxins and
viruses. Agglutination and precipitation reactions take place
in vitro, although it is not known how significant a role these
play in vivo.
 Each IgG subclass, has a unique biological property.
 Example, IgG1 and IgG3, supports phagocytosis because
they have macrophage receptor on the FC fragment of the
IgG.
 Protein A: protein from S. aureus.
 Placental transport to Fetus: IgG2 cannot be able to cross
the placenta.
 Again, analyze the table and memorize its content as the
discussion is the same on what is shown above.
IMMUNOGLOBULIN M (IGM)
 Known as a macroglobulin.
 With half-life of IgM is about 10 days-much shorter than that
of IgG.
 It accounts for between 5 and 10 percent of all serum
immunoglobulins.
 The pentamer form is found in secretions, while the
monomer form occurs on the surface of B cells.
 Found mainly in the intravascular pool and not in other body
fluids or tissues. It cannot cross the placenta.
 IgM is known as the primary response antibody, because it
is the first to appear after antigenic stimulation, and it is the
first to appear in the maturing infant.
CARIAGA, DELA CRUZ. GALANTO
 IgG is better at precipitation reactions than at agglutination,
because precipitation involves small soluble particles,
which are more easily brought together by the relatively
small IgG molecule. Agglutination is the clumping together
of larger particles such as red blood cells, and being a larger
molecule, IgM is much more efficient at this than IgG.
o Opsonin coats the antigen and bring to the
macrophage by way of the FC receptor, binding to the
FC receptor of IgG.
o Precipitation: the combining of small soluble particles.
o Agglutination: the clumping of larger particles, forming
a lattice formation .
o IgG and IgM is first to produce if you are first infected.
But the first to appear in the serum is IgM. But later on,
during the primary infection, you are already producing
IgG
 It is synthesized only as long as antigen remains present,
because there are no memory cells for IgM.
 Functions of IgM include (1) complement fixation, (2)
agglutination, (3) opsonization, and (4) toxin neutralization.
o IgM can fix the complement pathway, considering that
it is big and it is a macroglobulin.
 IgM is the most efficient of all immunoglobulins at triggering
the classical complement pathway because a single
molecule can initiate the reaction as a result of its multiple
binding sites. This probably represents the most important
function of IgM.
 The larger number of binding sites also makes IgM more
efficient at agglutination reactions, especially with
multivalent antigens. Thus, IgM forms a potent defense
against many bacterial diseases.
 Because IgM has a J chain, it can occasionally acquire a
secretory component like IgA does, and this allows it to
traverse epithelial cells and patrol mucous membranes.
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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY
 IgM structure.
 Refer to the labels in the illustration.
 It is the most efficient of all Ig because it has many binding
sites compared to other Ig.
 IgM has low affinity (how strong an antibody binds with a
specific epitope or antigen). However, it is high avidity (it
has many binding sites).
 Figure B: conformational spatial
 Graph illustrating on primary response of infection to the
secondary response of infection.
 Refer to the labels in the illustration.
 For example, first exposure to an immunogen / antigen,
Dengue.
o You are not yet exposed before.
o Black line represents your IgM concentration in the
blood, while the Red Line represents your IgG
concentration in the blood.
 Lag phase: is the period between the infection and
production of the antibody, which is long since it is still
processing.
 Primary compared to secondary, has a longer lag phase.
Take note that that first Ab to be produced in the primary
response is the IgM because initially in the B-cell, that is
what is present.
CARIAGA, DELA CRUZ. GALANTO
 Later on, during its period, there is class switching so that
IgG is what is produced in an incremental manner during
the primary response.
 It is therefore clear that even if it is your first infection, you
are already producing both IgM and IgG. You don’t only
produce IgG during secondary response, but instead on
primary.
 Memory Cell: particular cell that causes shorter lag phase
for secondary response.
o Take note, that there is no memory B-cell for IgM, but
only on IgG. That is why during the secondary
response, IgG is higher in levels.
IMMUNOGLOBULIN A (IGA)
 In the serum, a monomer IgA represents 10 to 15 percent
of all circulating immunoglobulin,
 There are two sub-classes, designated IgA1 and IgA2.
They differ in content by 22 amino acids, 13 of which are
located in the hinge region and are deleted in IgA2.
 Lack of this region appears to make IgA2 more resistant to
some bacterial proteinases that are able to cleave IgA1.
Hence, IgA2 is the predominant form in secretions at
mucosal surfaces, while IgA1 is mainly found in serum.
 IgA2 is found as a dimer along the respiratory, urogenital,
and intestinal mucosa, and it also appears in milk, saliva,
tears, and sweat.
 Since mucosal surfaces are a major point of entry for
pathogens, IgA2 serves to keep antigens from penetrating
further into the body.
 Secretory IgA is synthesized in plasma cells found mainly
in mucosal-associated lymphoid tissue, and it is released in
dimeric form. IgA is synthesized at a much greater rate than
that of IgG-approximately 3 grams per day in the average
adult-but because it is mainly in secretory form, the serum
concentration is much lower.
 Main function of secretory IgA is to patrol mucosal surfaces
and act as a first line of defense by neutralizing toxins
produced by microorganisms, and by preventing bacterial
adherence to mucosal surfaces. This prevents pathogens
from colonizing the mucosal epithelium. Since IgA is found
in breast milk, breastfeeding helps to maintain the health of
newborns.
 It appears that IgA is not capable of fixing complement by
the classical pathway, although aggregation of immune
complexes may trigger the alternate complement pathway.
 Lack of complement activation may actually assist in
clearing antigen without triggering an inflammatory
response, thus minimizing tissue damage.
 Neutrophils, monocytes, and macrophages possess
specific receptors for IgA. Binding to these sites triggers a
respiratory burst and degranulation. This occurs for both
serum and secretory IgA, indicating that they are capable
of acting as opsonins.
 The success of oral immunizations such as the Sabin
vaccine, which induces IgA almost exclusively,
demonstrates the effectiveness of IgA's protective role on
mucosal surfaces.
o Vaccination is not just for IgG that is being produced
(common notion).
o For Sabin Vaccine, it is IgA that is being produced.
o Oral route: effective route for Polio infection.
IMMUNOGLOBULIN D (IGD)
 IgD was not discovered until 1965, when it was found in a
patient with multiple myeloma.
o Bence-Jones Protein: detected in urine for multiple
myeloma.
 It is extremely scarce in the serum, representing less than
0.001 percent of total immunoglobulins.
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 TRANS: BASIC CONCEPTS OF IMMUNOLOGY

 It is synthesized at a low level and has a half-life of only 2

to 3 days.
 Most of the IgD present is found on the surface of
immunocompetent but unstimulated B lymphocytes.
 It is the second type of immunoglobulin to appear (IgM
being the first), and it may play a role in B-cell activation.
 The high level of surface expression and its intrinsic
flexibility make it an ideal early responder to antigen. Those
cells bearing only IgM receptors appear incapable of an IgG
response, while those with both IgM and IgD receptors are
capable of responding to T-cell help and switching to
synthesis of IgG, IgA, or IgE. Thus, IgD may play a role in
regulating B-cell maturation and differentiation.
 Because of its unusually long hinge region, IgD is more
susceptible to proteolysis than other immunoglobulins. This
may be the main reason for its short half-life.
 In the secreted form in the serum, IgD does not appear to  Mast cells does not create IgE.
serve a protective function, because it does not bind  One theory is that, Basophil is produced form the HSC of
complement, it does not bind to neutrophils or the bone marrow and undergone processes, eventually
macrophages, and it does not cross the placenta.
turned into mast cells.
 Another theory is from HSC, turned into mast cells.
IMMUNOGLOBULIN E (IGE)
o Basophil: blood form
 The least abundant immunoglobulin in the serum,
o Mast cell: tissue form
accounting for only 0.0005 percent of total serum
 Refer to the illustration: sensitized mast cell with bound IgE.
immunoglobulins.
Once exposed to allergen, in order for mast cell to be
 IgE is the most heat-labile of all immunoglobulins; heating
activated, the Ag must be able to cross link two antibodies
to 56 degrees C for between 30 minutes and 3 hours results
together.
in conformational changes and loss of ability to bind to
o Cross linking result: mast cell degranulation.
target cells.
Chemotactic factors later on attract neutrophils and
 IgE does not participate in typical immunoglobulin reactions other effector cells for inflammation to begin causing
such as complement fixation, agglutination, or rashes or secretions during allergic reactions.
opsonization, it is incapable of crossing the placenta.
 Instead, shortly after synthesis, it attaches to basophils and
tissue mast cells by means of specific surface proteins,
termed high-affinity FC R1 receptors, which are found REFERENCES
exclusively on these cells. The molecule binds at the CH3
domain on the FC region. This leaves the antigen-binding Notes from the discussion by Prof. Jordan M. Callueng, RMT,
sites free to interact with specific antigen. MD
o Effector cells like Tissue Mast Cells
 Plasma cells that produce IgE are located primarily in the
lung and in the skin.
o Diseases such as asthma, skin allergy, etc. Another big challenge, yet you overcame. Things will never be
 Mast cells are also found mainly in the skin and in the lining easy but the moment you have read this one, this calls for a
“mini” celebration of some sort, because you are able to survive
of the respiratory and alimentary tracts. One such cell may
and to learn these concepts that you will be needing ahead.
have several hundred thousand receptors, each capable of
binding an IgE molecule. It may not be a full transcript of discussion, but we hope could
 May serve a protective role by triggering an acute help.
inflammatory reaction that recruits neutrophils and
eosinophils to the area to help destroy invading antigens We may have different ways and means of learning the subjects,
that have penetrated IgA defenses. but we are one in mission to be a help to one another.
 Eosinophils, especially, play a major part in the destruction
of large antigens such as parasitic worms that cannot be Best of luck, future RMT!
easily phagocytized.
#LetLeniLead #LeniKiko2022 :>

CARIAGA, DELA CRUZ. GALANTO CSU BS MLS 3B 12