Complement System

Year III
Introduction
• The complement system is a series of serum proteins (9 factors) which, through
sequential proteolysis, nonspecifically increase immunity to infectious organisms
and proteins.

• The initial, inactive complement components are named C1 – C9

• Each complement component is synthesized in an “inactive” form.

• Once initial activation occurs, the inactive complement component is split into
fragments, designated by letters (e.g. C1q, C3a,)
Complement Components
• C1(C1q, C1r, C1s ) • Factor B
• C2(C2a, C2b) • Factor D
• C3(C3a, C3b) • CD35
• C4(C4a, C4b) • CD55
• C5(C5a, C5b) • Factor H
• C6 • Factor I
• C7 • DAF
• C8 • CR1
• C9
Definitions
• C-activation: alteration of C proteins such that they interact with the next
component

• C-fixation: utilization of C by Ag-Ab complexes

• C-inactivation: denaturation (usually by heat) of an early C-component resulting

in loss of hemolytic activity

• Convertase/esterase: altered C-protein which acts as a proteolytic enzyme for

another C-component
Complement Activation

• The complement system may be activated in 3 distinct ways, named:

• The Classical Pathway - activated by
• antigen/antibody complexes
• The Alternative Pathway- activated by
• Microbial cell walls
• (The Lectin Pathway - activated by
• Bacterial lectins.

• The end result of this activation is always the same.

Pathways of Complement Activation
CLASSICAL LECTIN ALTERNATIVE
PATHWAY PATHWAY PATHWAY

antibody antibody
dependent independent

Activation of C3 and
generation of C5 convertase

activation
of C5
LYTIC ATTACK
PATHWAY
Proteins of Complement System
• C1(qrs), C2, C3, C4, C5, C6, C7, C8, C9

• Factors B, D, H and I, properdin (P)

• Mannose Binding Lectin (MBL), MBL associated serine proteases (MASP-1

MASP-2)

• C1 inhibitor (C1-INH, serpin), C4-binding protein (C4-BP), decay accelerating

factor (DAF),

• C1 receptor (CR1), protein-S (vitronectin)

Activation Product of Complement Proteins
• Activated component are usually over-lined: e.g. C1qrs

• When enzymatically cleaved, the larger moiety, binds to the activation

complex or membrane and the smaller peptide is released in the
microenvironment

• Letter “b” is usually added to the larger, membrane-binding, peptide and “a”
to the smaller peptide (e.g., C3b/C3a, C4b/C4a, C5b/C5a), except C2 (the
larger, membrane-binding moiety is C2a; the smaller on is C2b)
Activation Product of Complement Proteins

• The ultimate end-result of complement activation is the formation of

the Membrane Attack Complex (MAC), formed by components C5
through C9, which literally punches holes in membranes

• In addition, intermediate products such as C3a, C5a, and C3b may

play a role in upregulating the immune response through chemotaxis
or opsonization.
Functions

• Opsonization to enhance phagocytosis

• Phagocyte attraction and activation

• Lysis of bacteria and infected cells

• Regulation of antibody responses

• Clearance of immune complexes

• Clearance of apoptic cells

Classical Pathway
• Initiation of this pathway requires the formation of antibody-antigen complexes.

• The classical pathway of complement activation begins when C1 is cleaved due to

interactions with Fc regions of IgM or IgG complexed with antigen.

• Two IgG1, IgG2 or IgG3 molecules are required for C1 binding, whereas a single
IgM is able to bind C1.

• The cascade continues as C2 and C4 are cleaved, yielding a new protease complex
which cleaves C3 to yield C5 convertase
Components of the Classical Pathway

C3 C4

C1 complex
Classical Pathway Generation of C3-convertase
Classical Pathway Generation of C3-convertase

C4b2a is C3 convertase

C4b
Classical Pathway Generation of C5-convertase

C4b2a3b is C5 convertase; it leads into

the Membrane Attack Pathway

C3 b
C4b
Alternative Pathway
• C1 is not involved
• Microorganisms can activate the C3bBb convertase to generate large amounts of
C3 cleavage products by stabilizing the enzyme on their (CHO) surfaces.(there by
protecting the C3b from factor H)
• It can also be activated by aggregated IgA and by substances found in the cobra
venom.
• Another protein, properdin (Factor P) acts subsequently on this bound convertase
to stabilize it further.
Alternative Pathway
• C3 is cleaved in three ways:
• C3 convertase of classic pathway,
• C3 convertase of alternative pathway
• C3 spontaneously breaking down at low rate to C3a and C3b.

• Microbial surfaces directly activate the interaction of Factor D with its substrate
Factor B.

• Once the C3 convertase is formed (as C3bBb) the addition of an additional C3b
molecule will result in formation of the C5 convertase.

• This will result in conversion of the C5-9 MAC.

C3-activation The Amplification Loop

If spontaneously-generated C3b
is not degraded

C3b b C3 b
C3-activation The Amplification Loop

C3 b b C3b

C3b
C3-activation The Amplification Loop

b
C3 b C3b C3b

C3b
C3-activation The Amplification Loop

C3b C3b C3b

C3b
C3-activation The Amplification Loop

C3b C3b

C3b
Mannose-binding Lectin Pathway

C4b2a is C3 convertase; it will lead to the

generation of C5 convertase

MASP1

MBL
Complement Opsonization

• Complement interacts with the immune cells through specific cell

surface receptors, functionally similar to Fc Receptors on B cells.

• Interaction of complement with these receptors increases phagocytosis

and may increase immune reactivity (through C3b/CD21 and the
membrane Ig receptor)
Complement Regulation

• Because the complement system is a cascade, which is capable of self

initiation, a number of proteins exist which regulate the process.

• These include C1 inhibitor-prevent activation of C1 hence inhibiting

clasic pathway

• Factor H, Factor I,or DAF (Decay Accelerating Factor) expressed on

the surface of self cells and inhibits C3 convertase.
 C1qrs Breakdown

C1Inh
Control of Spontaneous C3 Activation via DAF

DAF prevents
C3b

the binding of
CR1
factor B to C3b Autologous cell membrane
Control of Spontaneous C3 Activation via DAF

DAF dislodges
b C3b
C3b-bound
CR1
factor Bb
Autologous cell membrane
Control of Spontaneous C3 Activation via CR1

C3b C3b

CR1 iC3b
CR1

Autologous cell membrane

Degradation Of Spontaneously Produced C3b

C3c C3c

C3b C3b

C3dg
iC3b C3dg
iC3b
C5-convertase of The Two Pathways

C5-convertase of the Alternative

C5-convertase of the Classical and
Pathway
lectin Pathways

C3b C3b
C3b
C4b
 Lytic Pathway

Generation of C5 convertase leads

to the activation of the
Lytic pathway
Components of The Lytic Pathway

C7
C6

C
9
Lytic Pathway C5-activation

C3b
C4b
Lytic Pathway Assembly of The Lytic Complex

C6

C7 b
Lytic Pathway Assembly Of The Lytic Complex

C6

C7 b
Lytic Pathway: Insertion of Lytic Complex into Cell Membrane

C6

C7 b

C C
9 9 9 C
C 9
C
9 C
9
C
9 C C
9 9
Biological Effects of C5a
Opsonization and Phagocytosis
Biological Properties of C-activation Products

Product Biological Effects Regulation

C2b
(prokinin) edema C1-INH

C3a (anaphylatoxin) mast cell degranulation; carboxy-

enhanced vascular peptidase- B
permeability; anaphylaxis (C3-INA)
Biological Properties Of C-activation Products

Product Biological Effects Regulation

C3b (opsonin) opsonization; phagocyte factors H & I
activation

C4a (anaphylatoxin) as C3, but less (C3-INA)

potent

C4b (opsonin) opsonization; C4-BP, factor I

phagocytosis
Biological properties of C-activation products

Product Biological Effects Regulation

C5a (chemotactic anaphylactic as C3, but much carboxy-
factor) more potent; peptidase-C
attracts & activates PMN causes (C3-INA)
neutrophil aggregation,
stimulation of oxidative
metabolism and leukotriene
release

C5b67 chemotaxis, attaches to protein-S

other membranes
Summary

• Opsonization (via C3b)

• Inflammation (via C3a & C5a)

• Lysis ( via C5-9)