The Complement System

By Dr. B.P. Nayak

Position of Complements in
Innate Immune Response
• Inflammatory response enhances phagocytosis through
acute phase proteins
• Mannose-binding lectin (MBL)
• Binds to bacterial surface with particular spatial arrangement of
mannose or fucose
• C-reactive protein (CRP)
• Binds to phosphorylcholine on bacterial surface
• Complement
• Set of proteins which bind to bacterial surface
• Inflammatory response
• Accumulation of fluid and cells at infection site
(swelling, redness, heat and pain)
Position of Complements in
Adaptive Immune Response
• Creates millions of different B and T cells for specific
antibody-mediated and cell-mediated immunity
• Antibody-Mediated Immunity (AMI)
• Involves B lymphocytes, plasma cells and antibodies
• Humoral immunity
• Name derives from antibodies found in body fluids (humors - old medical term)
• Cell-Mediated Immunity (CMI)
• Involves T lymphocytes, antigen-presenting cells and MHC (major
histocompatibility complex) molecules
• Cellular immunity
 ADCC
Cross-linking of Fc
Antibody binds Fc receptors on receptors signals the Target cell dies by
antigens on the NK cells recognize NK cell to kill apoptosis and
surface of target cells bound antibody the target cell membrane damage

Around 1960-70, it was found that, ADCC is possible without the help of NKs
Then, HOW ?
The complement system

 A bridge between innate and adaptive immunity.

 It initiates a robust innate immunity (innate detection and

elimination of pathogenic infections),

 It can effectively modulate the adaptive immune

responses through regulation of interplay between B and
T cells
The complement system

 Complement refers to a complex set of 14 distinct serum

proteins (nine components) + 12 surface proteins that are
involved in three separate pathways of activation.

 Complement proteins designated by “C” followed by

numbers and letters.
General Properties
General Properties

 Primary role is cell lysis.

 Activity of complement destroyed by heating sera to 560 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
The complement system:
Mechanism of Action

• A defensive system consisting of over 30 proteins

produced by the liver and found in circulating blood serum.
• Complement kills microbes in three different ways
1. Opsonization
2. Inflammation
3. Cytolysis
How it Works?
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
THREE Pathways
• The complement pathway can be activated by either of three
different pathways.
The Classical Pathway

• C1 is a complex of 5 subunits

• C1 becomes activated when it

binds to the ends of antibodies

• It has intrinsic Serine protease

activity
The Classical Pathway
The building of a C3 activation complex

b a
C3 Activation complex

C3 activation complex
a OR
C3 convertase
OR
C4b2a
C3b
• Many C3b molecules are produced by the C3 activation
complex. (200:1)
• 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
1,000 fold.

Opsonins
C3a
C3a

C3a increases the inflammatory response by binding to

mast cells and causing them to release histamine
Building the C5 activation complex
C5 activation complex
OR
C5 convertase
OR
C4b2a3b
C5 activation complex
The C5 activation complex
• The C5 activation complex
(C4b2a3b) activates C5
proteins by cleaving them
into C5a and C5b

• Many C5b proteins are

produced by the C5
activation complex.

• These C5b begin to coat

the surface of the bacteria.
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
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

OR
MAC
OR
C5b6789
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
The alternative 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
 Virally infected cells
 Rabbit erythrocytes
The
Thealternative
alternativepathway
pathway

 Molecules of C3 undergo spontaneous cleavage at low level in

normal plasma.
 At least 4 serum proteins play important role.
 C3b attaches to appropriate site (activating surface) which is
actually a protective surface
 Progress to C3 activator stage without participation of C1, C4 or
C2.
 Activation sequence:
C3, C5, C6, C7, C8, C9.
Initiation: The alternative pathway

• C3 contains in unstable thioester bond.

• This unstable bond makes C3 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
Factor B and C3 convertase
Factor B has affinity for hydrolyzed C3 in plasma and
C3b on the surface of a foreign cells.
Factor D D
Factor

• Factor D has affinity for

C3bBb complex and cleaves
Factor B to Ba and Bb.

• Factor Bb remains bound to

C3b while Ba and Factor D
disperse away.

C3 activation complex
OR
C3 convertase
OR
C3bBb
TheThe
C3 C3 activation
activation Complex
complex

• The C3 activation complex (C3bBb) 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
Satbilization of C3 activation
complex

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

complex to stabilize it.
• C3bBbP make up the stable C3 activation complex for the
alternative pathway
C5 C5 activation
activation complex
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

• Activation of the lectin pathway begins when mannose-binding

protein (MBP) binds to the mannose groups of microbial
carbohydrates.
• Two more lectin pathway proteins called MASP1 and MASP2
(equivalent to C1r and C1s of the classical pathway) bind to the
MBP.
• This forms an enzyme similar to C1 of the classical complement
pathway that is able to cleave C4 and C2 to form C4bC2b, the C3
convertase capable of enzymatically splitting hundreds of
molecules of C3 into C3a and C3b
Lectin Pathway
LECTIN PATHWAY

 The beneficial results are the same as in the classical

complement pathway:
 Trigger inflammation (C5a >> C3a > C4a);
 Chemotactically attract phagocytes to the infection site (C5a);
 Promote the attachment of antigens to phagocytes via enhanced
attachment or opsonization (C3b > C4b >> C5b);
 Serves as a second signal for the activation of naive B-lymphocytes
(C3d);
 Cause lysis of gram-negative bacteria and human cells displaying
foreign epitopes (MAC).
 And remove harmful immune complexes from the body (C3b >
C4b).
Overview
Regulation: Complement Cascade

 Modulating mechanisms are necessary to regulate complement

activation and control production of biologically active split
products
 First type 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.
 Second type of control involves specific control proteins
 C1 inhibitor blocks activity of C1r and C1s.
 Factor H and I, activators in the presence of certain cofactors
inactivates C3b and C4b.
 A number of proteins act to control membrane attack unit
Regulation: Complement Cascade