Complement: History

Discovered in 1894 by Bordet

It represents lytic activity of

fresh serum

Its lytic activity destroyed

when heated at 56C for 30
min
 Complement functions

• Host benefit:
– opsonization to enhance phagocytosis
– phagocyte attraction and activation
– lysis of bacteria and infected cells
– regulation of antibody responses
– clearance of immune complexes
– clearance of apoptotic cells

• Host detriment:
– Inflammation, anaphylaxis
 Proteins of the complement
system (nomenclature)
• 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), Complement receptor 1 (CR1), protein-
S (vitronectin)
 Definitions

• C-activation: alteration of C proteins such that they

interact with the next component
• C-fixation: utilization of C by Ag-Ab complexes
• Hemolytic units (CH50): dilution of serum which
lyses 50% of a standardized suspension of Ab-coated
r.b.c
• 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
 Activation product of complement proteins
(nomenclature)

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 one is C2b)
 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
 Components of the Classical
Pathway

C1r C1s
Ca++
C1q
C2 C3 C4

C1 complex
 Classical Pathway
Generation of C3-convertase

C4a C1r C1s

Ca++

C4 b
C1q
 Classical Pathway
Generation of C3-convertase

C4a C1r C1s C2b

2 a
Ca++ C
C1q
_____
Mg++
C4b2a is C3 convertase

C4b C2a
 Classical Pathway
Generation of C5-convertase

C4a C1r C1s C2b C3a

Ca++ ________

C1q C4b2a3b is C5 convertase;

it leads into the Membrane
Mg++ Attack Pathway

C3
b
C4b C2a
Biological Activities of Classical Pathway
12
Components
Component Biological Activity

C2b Prokinin; cleaved by plasmin to yield kinin, which

results in edema
C3a Anaphylotoxin; can activate basophils and mast
cells to degranulate resulting in increased vascular
permeability and contraction of smooth muscle cells,
which may lead to anaphylaxis
C3b Opsonin
Activation of phagocytic cells
C4a Anaphylaotoxin

C4b Opsonin
 Control of Classical Pathway Components
13

Component Regulation

All C1-inhibitor (C1-INH); dissociates C1r and C1s from

C1q
C3a C3a-inactivator (C3a-INA; Carboxypeptidase B)

C3b Factors H and I; Factor H facilitates the degradation

of C3b by Factor I
C4a C3a-INH
C4b C4 binding protein (C4-BP) and Factor I; C4-BP
facilitates degradation of C4b by Factor I; C4-BP
also prevents the association of C2a with C4b thus
blocking formation of C3 convertase
C1-inhibitor deficiency:
hereditary angioedema
Components of mannose-binding
lectin pathway

C4
MASP2

MBL C2 MASP1
 Mannose-binding lectin pathway

_____
C4a C2b
C4b2a is C3 convertase; it
will lead to the generation of
C5 convertase
MASP1 C4b
C4
MASP2 2a
CC2
MBL C4b C2a
 Components of the
alternative pathway

C3 B
P
 Spontaneous C3 activation

Generation of C3 convertase
D
H2O

Bb
C3 i C3b

C3a

C3iBb complex has a very short half life

 C3-activation
the amplification loop

If spontaneously-generated
C3b is not degraded

C3a C3b Bb C3 b
 C3-activation
the amplification loop

C3 b Bb C3b

C3a C3b Bb
C3a
 C3-activation
the amplification loop
D

C3 b Bb Bb C3b
C3b

C3a C3b Bb
C3a
C3a
 C3-activation
the amplification loop

C3b Bb Bb C3b
C3b

C3a C3b Bb
C3a
C3a
 C3-activation
the amplification loop

Bb Bb C3b
C3b

C3a C3b Bb
C3a
C3a
 Control of spontaneous
C3 activation via DAF

DAF prevents
C3b
the binding of B

DAF
CR1
factor B to C3b Autologous cell membrane
 Control of spontaneous
C3 activation via DAF

DAF dislodges
Bb Bb C3b
C3b-bound

DAF
CR1
factor Bb Autologous cell membrane
 Control of spontaneous
C3 activation via CR1

Bb
H

Bb C3b
C3b
I I

DAF
DAF

iC3b iC3b
CR1
CR1
Autologous cell membrane
Degradation of spontaneously
produced C3b

C3c C3c
C3b
I C3b
I
C3dg
iC3b C3dg
iC3b
 C3b stabilization and
C5 activation
C3a
C3b finds an activator
(protector) membrane
This is stable C5 convertase
P D of the alternative pathway

Bb
C3b C3 b
 C5-convertase of the two
pathways

C5-convertase of the C5-convertase of the

Classical and lectin Alternative Pathway
Pathways

C3b C3b Bb C3b

C4b C2a
 Lytic pathway

Generation of C5 convertase
leads to the activation of the

Lytic pathway
 Components of the lytic pathway

C7
C5 C6

C8
C
9
Lytic pathway
C5-activation

C5a

C5 b

C3b
C4b C2 a
 Lytic pathway
assembly of the lytic complex

C6

C7 C5 b
 Lytic pathway:
insertion of lytic complex into cell membrane

C6

C8 C5 b
C7
CC C C
C9 9 9 9C
9C C C9
9 9 9
Biological effects of C5a
 Biological properties of C-activation
products

Product Biological Effects Regulation

C2b
(prokinin) edema C1-INH

C3a mast cell degranulation; carboxy-

(anaphylatoxin) enhanced vascular peptidase- B
permeability; (C3-INA)
anaphylaxis
 Biological properties of C-activation
products

Product Biological Effects Regulation

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

C4a as C3, but less (C3-INA)

(anaphylatoxin) potent

C4b opsonization; C4-BP,

(opsonin) phagocytosis factor I
 Biological properties of C-activation
products

Product Biological Effects Regulation

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

C5b67 chemotaxis, attaches protein-S

to other membranes
 Complement Deficiencies and Disease 39
Classical Pathway

Pathway Component Disease Mechanism

C1INH Hereditary Overproduction of C2b

Angioedema (prokinin)

C1, C2, C4 Predisposition Opsonization of immune

to SLE complexes help keep
them soluble, deficiency
results in increased
precipitation in tissues
and inflammation
 Complement Deficiencies and Disease 40
Lectin Pathway

Pathway Component Disease Mechanism

MBL Susceptibility to Inability to initiate

bacterial infections lectin pathway
in infants or
immunosuppressed
 Complement Deficiencies and Disease 41
Alternative Pathway

Pathway/Component Disease Mechanism

Factors B or D Susceptibility Lack of sufficient

to pyogenic opsonization of bacteria
(pus-forming)
bacterial
infections
C3 Susceptibility Lack of opsonization and
to bacterial inability to utilize the
infections membrane attack pathway
C5, C6, C7 C8, or Susceptibility Inability to attack the outer
C9 to Gram- membrane of Gram-
negative negative bacteria
infections
 Complement Deficiencies and Disease 42
Alternative Pathway cont.

Pathway Component Disease Mechanism

Properdin (X-linked) Susceptibility Lack of opsonization of

meningococcal bacteria
meningitis

Factors H or I C3 deficiency Uncontrolled activation of

and C3 via alternative
susceptibility to pathway resulting in
bacterial depletion of C3
infections