IMMUNOGENS

and ANTIGENS
• IMMUNOGENICITY - The capacity of a substance to induce
an immune response under a given set of conditions

• IMMUNOGENS - Agent capable of binding immune receptors

and inducing an immune response by B cells and T cells

• ANTIGENS - Agent that binds with varying degrees of

specificity to immune receptors (antibodies on B cells; T cell
receptor on T cells)
EPITOPES RECOGNIZED BY
T OR B CELLS
 MAJOR CLASSES OF
ANTIGENS/IMMUNOGENS
a. Proteins or glycoproteins
b. Carbohydrates or polysaccharides
c. Lipids
d. Nucleic acids
 REQUIREMENTS FOR
IMMUNOGENICITY
a. Size, dose, route
b. Chemical composition
c. Foreignness
d. Adjuvants/Degradability
 HAPTENS

• low molecular weight compounds that are non-

immunogenic by themselves but become
immunogenic after conjugation to high molecular
weight carrier substances that are immunogenic
 ADJUVANTS

• An adjuvant is a substance, which when mixed with

an immunogen, enhances the immune response
against the immunogen
 Examples of Immunologic
Adjuvants
a. Freund’s Complete adjuvants
b. LPS
c. Muramyldipeptide
d. Systemic polynucleotide
e. Aluminum hydroxide
f. Cytokines
The Complement
system
 Introduction

• Components numbered in order of discovery.

• Sequence of activation is not in numerical
order.
• Components circulate in inactive precursor
form, develop activity upon activation.
• Complement proteins designated by “C” followed
by numbers and letters.
 The complement system

• refers to a set of serum proteins that cooperates with

both the innate and the adaptive immune systems to
eliminate blood and tissue pathogens.
• Complement kills microbes in three different ways
• 1. opsonization
• 2. inflammation
• 3. Cytolysis - MAC
• Paul Ehrlich coined the term “complement” as the
activity of blood serum that completes the action of
antibody
• Most components are synthesized in the liver
 General Properties of Complement

• Primary role is cell lysis.

• Activity of complement destroyed by heating sera to 56 C for
30 minutes.
• IgM and IgG are the only immunoglobulin capable of
activating complement (classical pathway).
• Complement activation can be initiated by complex
polysaccharides or enzymes (alternative or properdin pathway).
• Portions of the complement system contribute to chemotaxis,
opsonization, immune adherence, anaphylatoxin formation,
virus neutralization, and other physiologic functions
 FUNCTIONS OF THE
COMPLEMENT SYSTEM
a. Cytolysis of foreign organisms (e.g. bacteria)
b. Opsonization and phagocytosis of foreign
organisms
c. Activation of inflammation and directed migration
of leukocytes
d. Solubilization and clearance of immune complexes
e. Enhancement of humoral immune response
 Functional Categories of the
Complement
A. Initiator complement components – C1q complex,
Mannose Binding Lectin (MBL) and ficolin
B. Enzymatic mediators – C1r, C1s, MASP2 and factor B
C. Membrane – binding components or opsonin – C3b,
C4b
D. Inflammatory mediators – C3a, C5a and C4a
 Functional Categories of the
Complement
E. Membrane attack proteins – C5b, C6, C7, C8 and C9
F. Complement receptor proteins – CR1, CR2 and C5aR
G. Regulatory complement components – factor I,
protectin
 The
complement
system
 A Cascade system

• The complement works as a cascade system.

• Cascade is when one reaction triggers another reaction
which trigger others and so on. These types of systems
can grow exponentially very fast.
 Cascade activation

• Complement proteins are often designated by an

uppercase letter C and are inactive until they are split
into products.
• Example: C1
• When the products are split they become active. The
active products are usually designated with a lower
case a or b.
• Example: C1a and C1b
 Two Pathways

• The complement pathway can be activated by either

of two different pathways.
• Classical pathway (specific immune system)
• alternative (non-specific immune system)
 The Classical Pathway
• The classical pathway is
considered to be part of the
specific immune response
because it relies on antibodies
to initiate it.

• C1 becomes activated when it

binds to the ends of
antibodies
 The building of a C3 activation complex

• Once C1 is activated, it
activates 2 other complement
proteins, C2 and C4 by cutting
them in half
• C2 is cleaved into C2a and
C2b
• C4 is cleaved into C4a and
C4b
• Both C2b and C4b bind
together on the surface of the
bacteria
• C2a and C4a diffuse away
 C3 Activation complex

• C2b and C4b bind

together on the surface
to form a C3 activation
complex

• The function of the C3

activation complex is to
activate C3 proteins.
• This is done by cleaving
C3 into C3a and C3b
 C3b

• Many C3b molecules are produced by the C3

activation complex.
• The C3b bind to and coat the surface of the
bacteria.
• C3b is an opsonin
• Opsonins are molecules that bind both to
bacteria and phagocytes
• Opsonization increases phagocytosis by Opsonins
1,000 fold.
 C3a

C3a increases the inflammatory response by

binding to mast cells and causing them to
release histamine
 Building the C5 activation complex

• Eventially enough C3b is cleaved that the surface of

the bacteria begins to become saturated with it.
• C2b and C4b which make up the C3 activation
complex has a slight affinity for C3b and C3b binds
to them
• When C3b binds to C2b and C4b it forms a new
complex referred to as the C5 activation complex
 The C5 activation complex

• The C5 activation complex (C2b, C4b, C3b) activates

C5 proteins by cleaving them into C5a and C5b

• Many C5b proteins are produced by the C5activation

complex. These C5b begin to coat the surface of the
bacteria.
 The function of C5a

• C5a disperses away from the bacteria.

• Binds to mast cells and increases inflammation.
• Most powerful chemotactic factor known for leukocytes
 Building the Membrane Attack complex

• C5b on the surface of bacteria binds to C6

• The binding of C6 to C5b activates C6 so that it can
bind to C7
• C7 binds to C8 which in turn binds to many C9’s
• Together these proteins form a circular complex
called the Membrane attack complex (MAC)
 Membrane Attack complex

• The MAC causes Cytolysis.

• The circular membrane attack
complex acts as a channel in
which cytoplasm can rush out
of and water rushes in.

• The cells inner integrity is

compromised and it dies
Overview
Membrane Attack Unit
 Order of Activation in Classical
Pathway

• C1, C4, C2, C3, C5, C6, C7, C8, C9

 Alternative Pathway
(Properdin Pathway)

• Cleavage of C3 and activation of the remainder of the

complement cascade occurs independently of
antibody.
• Triggers for the alternative pathway include
• bacterial cell walls
• bacterial lipopolysaccharide
• fungal cell walls
• some virus infected cells
• and rabbit erythrocytes
 The alternative pathway

• The alternative pathway is part of the non-specific

defense because it does not need antibodies to
initiate the pathway.

• The alternative pathway is slower than the Classical

pathway
The Alternative
complement pathway
 Initiation of The Alternative pathway

• C3 contains in unstable
thioester bond.
• This unstable bond
makesC3 subject to slow
spontaneous hydrolysis to
C3b and C3a
• The C3b is able to bind to
foreign surface antigens.
• Mammalian cells contain
sialic acid which inactivates
C3b
 Alternative Pathway
(Properdin Pathway)
• Action by the 4 serum proteins on C3b proceeds to the
C3 activator stage without participation of C1, C4 or
C2.
• Activation sequence:
C3, C5, C6, C7, C8, C9.
 Factor B

• C3b on the surface

of a foreign cells
binds to another
plasma protein called
factor B
 Factor D
• The binding of C3b to factor
B allows a protein enzyme
called Factor D to cleave
Factor B to Ba and Bb.

• Factor Bb remains bound to

C3b while Ba and Factor D
disperse away.
 The C3 activation complex

• Properdin, also called factor P, binds to the C3bBb complex to

stabilize it.

• C3bBbP make up the C3 activation complex for the alternative

pathway
 The C3 activation Complex
• The C3 activation complex
causes the production of
more C3b.
• This allows the initial steps of
this pathway to be repeated
and amplified
• 2X106 molecules can be
generated in 5 minutes
 C5 activation complex
• When an additional C3b
binds to the C3 activation
complex it converts it into a
C5 activation complex.

• The C5 activation complex

cleaves C5 into C5a and C5b.

• C5b begins the production of

the MAC.
Overview
 Lectin Pathway

• the most recently discovered mechanism for

complement activation
• When macrophages ingest bacteria, viruses, and
other foreign matter by phagocytosis, they release
cytokines that stimulate the liver to produce lectins,
proteins that bind to carbohydrates,
Overview
Solubilization and Clearance of
Immune Complexes
Overview
 Regulation of the Complement Cascade

• Modulating mechanisms are necessary to regulate

complement activation and control production of
biologically active split products
 Regulation of the Complement Cascade

• First means of control is extreme lability of activated

complement
• If activated complement does not combine within
milliseconds the activity is lost or decreased.
• Active fragments rapidly cleared from the body.
 Regulation of the Complement Cascade

• Second type of control involves specific control

proteins
• C1 inhibitor blocks activity of C1r and C1s.
• Factor I in activator in the presence of certain cofactors
inactivates C3b and C4b.
• A number of proteins act to control membrane attack unit
• C1INH inhibits both C3b and the serine protease MASP2
 Regulated by protein stability and
cell–surface composition
• relative instability of many complement components
is the first means by which the host protects itself
against inadvertent complement activation.
• second passive regulatory mechanism depends upon
the diff erence in the cell surface carbohydrate
composition of host versus microbial cells
Other Complement Inhibitors
Complement deficiencies
 Classical pathway deficiencies
result in tissue inflammation
• Deficiency of any of the C1 subunits (C1q, C1r,
or C1s) invariably causes severe disease with typical
SLE features including skin lesions and kidney
damage
• C4 deficiency also causes severe SLE
• C2 deficiency is the commonest complement
deficiency in Caucasians
 Deficiencies of MBL are associated
with infection in infants
• associated in infants with increased susceptibility to
bacterial infections
Alternative pathway and C3 deficiencies
are associated with bacterial infections

• Prevent complement amplification through the

alternative pathway amplification loop, markedly
reducing the efficiency of opsonization of pathogens
• Individuals with C3 deficiency present early in
childhood with a history of severe, recurrent
bacterial infections affecting the respiratory system,
gut, skin, and other organs.
Terminal pathway deficiencies
predispose to Gram-negative bacterial
infections
• Individuals with terminal pathway deficiencies usually
present with meningitis, which is often recurrent and
often accompanied by septicemia
• In Caucasians and most other groups investigated,
C6 deficiency is the most common;
• In the Japanese population, C9 deficiency is very
common, with an incidence of more than 1 in 500 of
the population.
Complement Deficiencies and
Associated diseases
 C1 inhibitor deficiency leads to
hereditary angioedema (HAE)
• Often triggered in the skin or
mucous membranes by minor
trauma
• Involves a mutation that prevents
synthesis of C1inh by the defective
gene
• In Type II HAE, the plasma levels
of C1inh may be normal or even
high, but its function is markedly
impaired
• Deficiency in alternative pathway regulators produce
a secondary loss of C3 predisposes to bacterial
infections and yields a clinical picture identical to
primary C3 deficiency
 Properdin deficiency causes severe
meningococcal meningitis
• Inherited in an X-linked manner
• The first attack is often fatal and survivor do not
usually have recurrent infection
 Assignment

1. Explain in details the different biological

functions mediated by complement
activation
2. Hoes does complement clear immune
complexes from the circulation?
3. Explain how the complement activation
pathways are affected if a certain
component is missing.