THE IMMUNE SYSTEM

The Invaders The Defender

COMPONENTS OF THE IMMUNE SYSTEM
Cellular components of the immune system

Lymphocytes Phagocytes Granulocytes Others

B-cell Mononuclear phagocyte Basophil Platelets

T-cell Neurtrophil Mast cell

Large granular lymphocyte Eosinophil

Antibodies Cytokines Complement Inflammatory mediators

Soluble mediators of the immune system

 ORIGINATION OF CELLULAR
COMPONENTS
Lymphoid
stem cell

T-cell B-cell Plasma Natural

cell killer cell

Pluripotent Stem cell

Neutrophil Monocyte Eosinophil Basophil Mast cell Platelets

Myeloid
Stem cell

Macrophage
THE EVOLUTION OF IMMUNITY

Immunity

Non-specific Immediate onset Specific Delay onset

Innate immunity Acquired immunity

Humoral Cellular
Immune Response Immune Response
Antibodies production T-cell activation
 INNATE IMMUNITY
Pathogenic invasion Bacteria Viruses
1st Line Defense

Skin Ciliated cells Lysozyme Coughing Vomitting

2nd Line Defense

Blood clot Mast cell Cytokines Leukocytes

THE INFLAMMATORY RESPONSE

Tissue injury caused by pathogenic invasion

Capillary Increased Leukocyte Systemic

widening capillary attraction response
permeability

Increased Release Leukocytic Leukocytic

blood flow of fluid migration proliferation

Heat, Redness, Tenderness, Swelling, Pain

THE COMPLEMENT CASCADE
Classical Alternative

Antibody binding Mannose binding

The Complement System

Opsonization Inflammation Cytolysis

ACQUIRED IMMUNITY
Humoral Immunity

Cytokines

Activated B-Cell Activation

TH cell Antibodies Production

Cytokines

Cytotoxic T-Cell Activation

Cell lysis

Cell-mediated Immunity
 ATTRIBUTES OF ACQUIRED
IMMUNITY
Immunological Specificity
Immunological Tolerance

Receptor
APC

Immunological memory

B-cell Activated T-cell

Plasma Memory Mature Memory

cell B-cell T-cell T-cell
THE COMPLEMENT
SYSTEM
 complement system
It is a part of the immune system that helps or
complements the ability of antibodies and phagocytic
cells to clear pathogens from an organism.
It is part of the innate immune system, which is not
adaptable and does not change over the course of an
individual's lifetime.
However, it can be recruited and brought into action
by the adaptive immune system.
It consists of a number of small proteins found in the
blood, in general synthesized by the liver, and
normally circulating as inactive precursors (
pro-proteins).
 complement system
When stimulated by one of several triggers, proteases in the
system cleave specific proteins to release cytokines and
initiate an amplifying cascade of further cleavages. The end-
result of this activation cascade is massive amplification of the
response and activation of the cell-killing
membrane attack complex.
Over 30 proteins and protein fragments make up the
complement system, including serum proteins, serosal
proteins, and cell membrane receptors.
They account for about 5% of the globulin fraction of blood
serum and can serve as opsonins.
Three biochemical pathways activate the complement system:
the classical complement pathway, the
alternative complement pathway, and the lectin pathway.
 Overview
The complement system is part of the
innate immune system (vs adaptive)
It is named complement system because
it was first identified as a heat-labile
component of serum that
complemented antibodies in the
killing of bacteria
It is now known that it consists of over 30
proteins and contributes 3 g/L to overall
serum protein quantities
 History of
complement
Ehrlich role of complementing
antibodies in killing of bacteria.
1895 Bordet
Subsequent discovery of components
Current knowledge:-
> 30 proteins in plasma + on cell surfaces
~ 15% of globulin fraction of proteins
 Nomenclature
C1 C1q, C1r, C1s
C4, C2, C3, C5, C6, C7, C8, C9
Many referred to as zymogens
a and b added in to denote cleavage
products.
b larger fragment
Alternative pathway proteins:- Factors or
identified by single letters
Complement receptors:- named according to
ligand (eg C6 receptor) or using CD system.
 Main roles
Defends against pyogenic bacterial
infections
Bridges both the innate and adaptive
immunity systems
Assists in disposing of immune
complexes etc
Role in Inflammation
1. Opsonization:- C3b
is important!
2. Chemotaxis:-
complement
fragments diffuse
from target
stimulating cellular
movement and
activation.
3. Target cell
lysis:-membrane
attack complex
hydrophobic plug
inserted into lipid
membrane bilayer
Activation
Pathways:-
1. Classical
2. Lectin
3. Alternative
Common end point: formation of C3
convertase cleaves to C3a and C3b
Classical + Lectin pathways C4b2a
Alternative pathway C3bBb
Ultimately:- converted into C5 convertase
by further addition of C3b. Production of
MAC.
1. Classical pathway
Antibody directed
Begins with C1
C1
Pentamolecule C1q fragment (6 domains) + 2
x C1r + 2 x C1s
Antibody binds to two or more of the six
domains (binds either IgG or IgM molecules)
C1 complex undergoes conformational change
Autocatalysis of C1r
C1s activation
Classical Pathway

Begins with antibody binding to a cell surface and ends

with the lysis of the cell
The proteins in this pathway are named C1-C9 (the order
they were discovered and not the order of the reaction)
When complement is activated it is split into two parts
a smaller of the two
B larger part and usually the active part (except with factor 2)
Remember 3 Key Words
ACTIVATION
AMPLIFICATION
ATTACK
Classical Pathway
ACTIVATION
C1q portion of C1 attaches to the Fc portion of
an antibody
Only IgG and IgM can activate complement
Once activated C1s is eventually cleaved which
activates C4 and C2
C4b & C2a come together to form the C4b2a
which is the C3 convertase
C3 convertase activates C3 to C3a and C3b
Classical Pathway
ACTIVATION
C3a binds to receptors on basophils and
mast cells triggering them to release there
vasoactive compounds (enhances
vasodilation and vasopermeability)
C3a is called an anaphylatoxin
C3b serves as an opsonin which facilitates
immune complex clearance
Classical Pathway

AMPLIFICATION
Each C1s creates many C4b and C2b fragments
Each C4bC2a creates many C3b (activated C3)
Each C3b goes on to create many Membrane
Attack Complexes
Example
1 C1S makes 100 C4bC2b
100 C4bC2b makes 10,000 C3b
10,000 C3b makes 1,000,000 MAC
Classical Pathway
ATTACK
Most C3b serves an opsonin function
Some C3b binds to C4bC2a to form the C5
convertase C4bC2aC3b
C5 convertase cleaves C5 leading to the
formation of the Membrane attack Complex
(C5-C6-C7-C8-C9)
The MAC punches holes in cell walls
resulting in lysis
 C5a is a: C3a binds to receptors on
C3 basophils and mast cells
C2
C4 1. Potent anaphylatoxin triggering them to release
C1q 2. Chemoattractant for there vasoactive compounds
neutrophils (enhances vasodilation and
vasopermeability) -
ANAPHYLATOXIN
C3a
C3b C5
2a
4
2b
4b
a
C5bC5a
C3-convertase
C5-Convertase

C7
C8
C9 C6

Classical
Pathway
2. Lectin pathway
Antibody independent
C1q calcium-dependent lectin (collectin)
Other members:- mannan-binding lectin (MBL),
conglutinin and lung surfactant A + D.
MBL may bind mannose grps on bacterial
surface then interacts with associated Serine
Proteases MASP1 and 2 (homologous to C1r
and C1s).
Antibody independent activation of classical
pathway
Downstream effects
C1 cleaves C4 forming activated C4b
Two isotypes exist
C4A binding amine grps (usually on proteins)
C4B hydroxyl grps on CHO
C4b allows binding of C2. Acted on by
C1s to release C2b.
C4b + C2a = classical pathway convertase
(C3)
By definition:- C3 convertase breaks up
C3 to C3a and C3b (focus of further
complement activation)
What about regulation?
C1 inhibitor serine proteinase inhibitor (aka
serprin) binds and inactivates C1r and C1s
Inhibition of formation of C3 convertase enzyme-
C4b2a (by ongoing catabolization of C4b by Factor
I and C4 binding protein)
Other complement control factors inhibit
complement binding to host cell surfaces
DAF: (Decay accelerating factor) CD55
CR1
MCP: Membrane co-factor protein
Inhibit binding of C2 to C4b; promote decay acceleration
of C2a from C4b. Assist in catabolism of C4b by Factor I
3.Alternative pathway
Spontaneous activation C3 is susceptible
to spontaneous hydrolysis by water
Tick over activation to form C3i
C3i acts as binding site for Factor B
(cleaved by Factor D to form Ba)
C3iBb alternative pathway C3
convertase
Most C3b generated becomes inactivated
in water. If it comes into contact with non-
self initiates amplification loop of
alternative pathway.
Regulation its always about
rules!!!
Factor H and I
DAF + CR1 accelerate dissociation of
C3bBb

How C3b reacts is governed by the

surface to which it attaches protected
vs non-protected
Alternative
Pathway
Requires no specific recognition of
antigen in order to cause activation
Alternative
Pathway
ACTIVATION
Spontaneous conversion from C3 to C3b
occurs in body
Normally, C3b is very short lived and quickly
inactivated by proteins on the surface of the
bodys own cell walls
However, bacteria or other foreign material
may lack these surface proteins allowing
C3b to bind and stay active
Alternative
Pathway
AMPLIFICATION
Factor B binds to C3b
Factor B is then cleaved by factor D into Ba
and Bb
C3bBb remains which acts as a C3
convertase (C3 C3a and C3b)
C3bBbC3b is formed which acts as a C5
convertase
Alternative
Pathway
ATTACK
C5 is cleaved to C5a and C5b
C5b then starts the assembly of the
Membrane Attack Complex
 C3a
C3
C3b

Anaphylatoxin C3
C5

Alternative
Pathway
C3-Convertase
C5-Convertase C3a C3b
C5b C5aD
C7
C8 BBa
Bb
C9 C6
Summary - Activation
Complement can be activated by the
binding of antibody (Classical) or by the
adherance of C3b to foreign material
(Alternative)
The two pathways converge at the
formation of the C5 convertase (C4b2a3b
or C3bBbC3b)
The final common pathway is the formation
of the membrane attack complex
Summary - Function

Opsonization C3b
Chemotaxis C5a (attracts neutrophils)
Increases vasodilation & permeability
of capillary beds via mast cell and
basophil activation C3a & C5a
(Anaphylatoxins)
Cellular Lysis via the MAC
 Initiators of complement
activation pathways
Classical
Immune complexes
Apoptotic cells
Viruses + GN bacteria
CRP bound to ligand
Lectin
Mannose groups terminal ends of
microbes
Alternative
Bacteria, fungi, viruses, tumour cells etc
 Membrane attack
complex
Requires enzymatic cleavage of C5
Sequential binding of C6, C7 (hydrophobic
status), C8, C9 (up to 14 monomers)
Formation of lytic plug majority of damage
caused by C9
C9 analogous to perforin (used by T
lymphocytes)
C5b67 can be inactivated by numerous means
(S protein vitronectin etc)
RBC immunity: poorly lysed by homologous
complement
CD59: glycophospholipid foot. Inhibits insertion +
unfolding of C9 into membranes.
Clinically speaking
CH50 / THC (total haemolytic complement):-
requires all nine components of classical
pathway to give normal value used to screen
for deficiency of classical pathway.
If very low - ? Homozygous deficiency of classical
pathway component
Less dramatic reduction during inflammatory process
AH50: alternative pathway measure
C3/4:- helpful as activity markers in those with
SLE
Anaphylatoxins:- C5a / C3a if increased
complement activation
 Elevated complement levels = inflammatory
response (i.e acute phase reaction) esp C3 /
C4 / B
Reduced levels: often a/w disease involving
immune complexes / autoantibodies. May be
useful for Dx + Mx of certain diseases (eg
SLE, Sjogrens, vasculitis etc)
Low C4 / C3 + N FB classical pathway
activation
Low FB + C3 + N C4 alternative pway
activation
C4 + FB low = both pways activated
Clinical implications
1. Complement deficiencies
2. Glomerulonephritis
3. C1 inhibitor deficiency
4. SLE
5. PNH
6. Sepsis
7. APLS
Complement deficiency:-
Increased susceptibility to pyogenic
infections
Contributing factors
Deficient opsonisation
Deficiency compromising lytic activity
Deficient manose-binding lectin pathway
1. Pyogenic infection:-
Site of defect:- antibody production, complement proteins of classical pathway,
phagocyte function
Usually bacteria is opsonised with Ab complement is then activated,
phagocytosis occurs and intracellular killing
Key player:- C3b
2. Impaired lysis
MAC component deficiency a/w Neisserial disease*
Risk of meningococcal disease ~ 0.5% / yr (RR 5000 cf normal population)
3. Deficient lectin
Deficiency occurs due to 1 of 3 point mutations a/w reduced levels.
Associated with higher risk of infection in children whilst losing passive
immunity
? Protective against mycobacterial infections
 IN SUMMARY OF CLASSICAL
PATHWAY
The classical pathway is triggered by activation of the C1-
complex.
The C1-complex is composed of 1 molecule of C1q, 2
molecules of C1r and 2 molecules of C1s, or C1qr2s2.
This occurs when C1q binds to IgM or IgG complexed with
antigens.
A single IgM can initiate the pathway, while several, ideally
six, IgGs are needed.
This also occurs when C1q binds directly to the surface of
the pathogen.
 Such binding leads to conformational changes in the
C1q molecule, which leads to the activation of two
C1r molecules. C1r is a serine protease.
They then cleave C1s (another serine protease). The
C1r2s2 component now splits C4 and then C2,
producing C4a, C4b, C2a, and C2b. C4b and C2b
bind to form the classical pathway C3-convertase
(C4b2b complex), which promotes cleavage of C3
into C3a and C3b; C3b later joins with C4b2b (the C3
convertase) to make C5 convertase (C4b2b3b
complex). The inhibition of C1r and C1s is controlled
by C1-inhibitor.
 IN SUMMARY OF THE ALTERNATE
PATHWAY
It is continuously activated at a low level as a result of spontaneous C3
hydrolysis due to the breakdown of the internal thioester bond (C3 is mildly
unstable in aqueous environment).
It does not rely on pathogen-binding antibodies like the other pathways.
C3b that is generated from C3 by a C3 convertase enzyme complex in the
fluid phase is rapidly inactivated by factor H and factor I, as is the C3b-like C3
that is the product of spontaneous cleavage of the internal thioester.
In contrast, when the internal thioester of C3 reacts with a hydroxyl or amino
group of a molecule on the surface of a cell or pathogen, the C3b that is now
covalently bound to the surface is protected from factor H-mediated
inactivation.
The surface-bound C3b may now bind factor B to form C3bB. This complex in
the presence of factor D will be cleaved into Ba and Bb.
Bb will remain associated with C3b to form C3bBb, which is the alternative
pathway C3 convertase.
 The C3bBb complex is stabilized by binding oligomers of factor P.
The stabilized C3 convertase, C3bBbP, then acts enzymatically to
cleave much more C3, some of which becomes covalently
attached to the same surface as C3b.
This newly bound C3b recruits more B,D and P activity and greatly
amplifies the complement activation. When complement is
activated on a cell surface, the activation is limited by endogenous
complement regulatory proteins, which include CD35, CD46, CD55
and CD59, depending on the cell.
Pathogens, in general, don't have complement regulatory proteins
(there are many exceptions, which reflect adaptation of microbial
pathogens to vertebrate immune defenses). Thus, the alternative
complement pathway is able to distinguish self from non-self on the
basis of the surface expression of complement regulatory proteins.
 Host cells don't accumulate cell surface C3b (and the proteolytic
fragment of C3b called iC3b) because this is prevented by the
complement regulatory proteins, while foreign cells, pathogens
and abnormal surfaces may be heavily decorated with C3b and
iC3b. Accordingly, the alternative complement pathway is one
element of innate immunity.
Once the alternative C3 convertase enzyme is formed on a
pathogen or cell surface, it may bind covalently another C3b, to
form C3bBbC3bP, the C5 convertase. This enzyme then cleaves
C5 to C5a, a potent anaphylatoxin, and C5b. The C5b then
recruits and assembles C6, C7, C8 and multiple C9 molecules to
assemble the membrane attack complex. This creates a hole or
pore in the membrane that can kill or damage the pathogen or
cell.
 IN SUMMARY OF THE LECTIN
PATHWAY
It is homologous to the classical pathway, but with the
opsonin, mannose-binding lectin (MBL), and ficolins,
instead of C1q.
This pathway is activated by binding of MBL to mannose
residues on the pathogen surface, which activates the
MBL-associated serine proteases, MASP-1, and MASP-2
(very similar to C1r and C1s, respectively), which can
then split C4 into C4a and C4b and C2 into C2a and C2b.
C4b and C2a then bind together to form the classical C3-
convertase, as in the classical pathway.
 Ficolins are homologous to MBL and function
via MASP in a similar way. Several single-
nucleotide polymorphisms have been described
in M-ficolin in humans, with effect on ligand-
binding ability and serum levels.
Historically, the larger fragment of C2 was
named C2a, but it is now referred as C2b.
Thank you for
listening