COMPLEMENT SYSTEM

HISTOR
Y
Research on complement began in the 1890s, when

Jules Bordet at the Institut Pasteur in Paris

showed that sheep antiserum to the bacterium Vibrio
cholerae caused lysis of the bacteria and that heating
the antiserum destroyed its bacteriolytic activity.
 He named those substances as

Alexins.
 Paul Ehrlich coined the term

complement.
 INTRODUCTION
 It is named “complement system” because it was first identified as

a heat-labile component of serum that “complemented or augment”

antibodies in the killing of bacteria.

 Consists of serum and cell surface proteins involved in

defense
against pathogens and tissue damage mediated by antibodies

 The Complement system is the major effector of cellular and

humoral branch of immune system.

 Plays major role in both innate and adaptive immunity.

 Complement system represents a group of about 30
proteins which augment or complement the immune
response.

 Most of these proteins are found in serum or on cell

surfaces.

 Synthesized in liver as inative precursors and are activated by

proteolysis durig their interaction in a sequential manner.

 Also produced by blood monocytes, tissue macrophages and

epithelial cells of he gastrointestinal and genitourinary
tract.
 General properties
Present in serum of all animals but its concentration
is maximum in serum of guinea pig.
Complement of one species are able to react with
antibodies of other species but not to the same extent.
C- proteins constitute about 5% of normal serum
protein
.
Are glycoproteins.
Are synthesized rapidly in inflammatory responses
– hence are called acute phase proteins.
Heat labile and lost activity at 56⁰ C for 30 mins and
inactivated. Immunoglobulins are not inactivated at this
temperature.
Binds with Fc potion of immunoglobulns .
Three main effects of complement are:

1. Lysis of cells (bacteria, allografts, tumor cells)

2. Generation of mediators of inflammation

3. Opsonization – enhancement of phagocytosis

 Complement system
 Over 30 serum and cell surface proteins:

 Complement components-

- Components are designated by numbers (E.g. ; C1 – C9) or latters

(E.g. : Factor D)
- (in serum inactive, activated sequentially as a cascade)

 Complement receptors

(cell surface, recognize activated components)

 Regulatory proteins of complement

(both in serum and cell surface, inhibit activated components)

 Complement proteins: are activation by cleavage.
proenzymes -

 Example: C4

C4a C4b

a = smaller fragment. b= larger fragment.

-Diffusion -remains bound to microbe

 Exception: C2: C2a = large fragment

C2b = small
fragment
 Complement Pathway
Three pathway of complement activation
1. Classical pathway:-
Is antibody dependent pathway and triggered by
formation of soluble antigen-antibody complex or by
binding of the antibody to the antigen present on
the target cell surface.

2. Alternative pathway:-
Is antibody independent pathway stimulated by
antigen directly eg. Bacterial cell surface components.

3. Lectin Pathway:-
Also antibody independent but resembles
classical pathway.
 Stages of complement Activation
Three main stages in the activation of complement
by any pathway are

 Formation of C3 convertage
 Formation C5 convertage
 Formation of membrane attack complec(MAC)

• The initiation and formation of C3 convertage are

different in classical and alternative pathway . These
then follow the parralel route to merge at C5 convertage
stage and finally generate the MAC by a common
route.
Sequential activation of complement components
occurs via one of three pathways:

 the classic pathway,

 the lectin pathway, and
 the alternative pathway .

Of these pathways, the lectin and the alternative

pathways are more important the first time we are
infected by a microorganism because the antibody
required to trigger the classic pathway is not present.
The lectin pathway and the alternative pathway are,
therefore, participants in the innate arm of the immune
system.
 Classical pathway
Part of acquired immunity.
In the classic pathway, antigen–antibody complexes
activate C1 to form a protease, which cleaves C2
and C4 to form a C4bC2a complex, C2a and C4b split
off.
The C4bC2a is C3 convertase, which cleaves C3
molecules into two fragments, C3a and C3b.
C3b forms a complex with C4b,2b, producing a new
enzyme, C5 convertase (C4b2a3b), which cleaves
C5 to form C5a and C5b
C5b binds to C6 and C7 to form a complex that
interacts with C8 and C9 to produce the membrane
attack complex (C5b,6,7,8,9), which causes
cytolysis.
 Note that the "b" fragment continues in the main
 Classic Pathway
Components of the Classical Pathway
Native component Active component(s) Function(s)
Binds to antibody that has bound
C1q
antigen, activates C1r.
C1(q,r,s) Cleaves C1s to activate protease
C1r
function.
C1s Cleaves C2 and C4.
C2a Unknown.
C2 Active enzyme of classical
C2b
pathway; cleaves C3 and C5.
Mediates inflammation;
C3a
anaphylatoxin.
C3 Binds C5 for cleavage by C2b.
Binds cell surfaces for
C3b opsonization and activation of
alternate pathway.
C4a Mediates inflammation.
C4 Binds C2 for cleavage by C1s.
C4b Binds cell surfaces for
opsonization.
Only IgM and IgG fix complement.
 One molecule of IgM can activate complement;
however, activation by IgG requires two cross-linked
IgG molecules.
Of the IgGs, only IgG1, IgG2, and IgG3 subclasses
fix complement; IgG4 does not.
C1 is bound to a site located in the Fc region of the
heavy chain.
C1 is composed of three proteins, C1q, C1r, and C1s.
 C1q is an aggregate of 18 polypeptides that binds
to the Fc portion of IgG and IgM.
It is multivalent and can cross-link
several immunoglobulin molecules.
C1s is a proenzyme that is cleaved to form an active
protease.
 Classical Pathway
Generation of C3-convertase
Generation of C3-
convertase

C4b2a is C3
convertase

C4b
 Alternative Pathway

• Also known as the Properdin Pathway

• Part of innate immunity

• Bypasses C1, C4, and C2

• Does not require an antigen-binding protein

• Does not wait for antibody to be formed for

activation

• Acts synergistically with the classical

pathway
 Alternative Pathway

• Usually activated by products of micro-

organisms like endotoxin

• Other activators include:

1. Complexes containing IgA
2. Some virus-infected cells (e.g. EBV)
3. Many gram negative and gram
positive
organisms
4. Parasites – Trypanosomes,
Leishmania
5. Dextran SO4
6. Erythrocytes
7. Carbohydrates (agarose)
 Alternative Pathway

Components of the Alternate Pathway

Active
Native component Function(s)
component(s)
Mediates inflammation;
C3a
anaphylatoxin.
C3 Binds cell surfaces for
C3b opsonization and activation
of alternate pathway.
Binds membrane bound
B
C3b. Cleaved by Factor D.
Factor B Ba Unknown.
Cleaved form stabilized by
Bb
P produces C3 convertase.
Cleaves Factor B when
Factor D D
bound to C3b.
Binds and stabilizes
Properdin P
membrane bound C3bBb.
 Lectin Pathway
 Also known as the MBL Pathway

In the lectin pathway, mannan-binding lectin (MBL)

(also known as mannose-binding protein) binds to the
surface of microbes bearing mannan (a polymer of
the sugar, mannose).
 Binding causes activation of MASP (MBP- associated
serine proteases)  cleave C2 and C4 and activate
the classic pathway.

Note that this process bypasses the antibody-

requiring step and so is protective early in infection
before antibody is formed.
Lectin Pathway
• All three pathways lead to production of C3b
 central molecule of complement cascade

• Presence of C3b on surface of a microbe

marks it as foreign and targets it for
destruction

• C3b with two important functions:

1. Combines with other complement
components to generate C5 convertase
2. Opsonizes bacteria
Components of the Membrane-Attack Complex
Active
Native component Function(s)
component(s)
Mediates inflammation;
C5a
anaphylatoxin, chemotaxin.
C5 Initiates assembly of the
C5b membrane-attack complex
(MAC).
Binds C5b, forms acceptor
C6 C6
for C7.
Binds C5b6, inserts into
C7 C7 membrane, forms acceptor
for C8.
Binds C5b67, initiates C9
C8 C8
polymerization.
Polymerizes around
C9 C9n C5b678 to form channel
that causes cell lysis.
 Classical Pathway
Generation of C5-convertase

C4b2a3b is C5
convertase; it leads
into the Membrane
Attack Pathway

C3
C4b
b
 C4b-2a-3b functions as the classical
C5 convertase:
 Lytic pathway
assembly of the lytic complex

C6

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

C6

C7 b

CCCC
C99 9
9C
9 CC C9
 Membrane attack complex
 Cleavage of C5 into C5a and C5b.
 C5 (structurally homologous to C3 and C4, lacks
internal thioester bond )
 C5b initiates formation of MAC (complex of C5b,
C6, C7, C8 and multiple C9 molecules ) binds to
C6, and C7 , recruits C8 and complex penetrates
more deeply into the membrane.
 C9, a pore-forming molecule with homology to
perforin. The complex of C5b678 forms a nidus for
C9 binding and polymerization
 Penetrates membrane bilayers to form pores
 Disrupt the osmotic barrier, leading to swelling and
lysis of susceptible cells Abbas et.al.Cellular&Molecular immunology 6th edition
Biologic Effects of complement:
C3b is an opsonin
Opsonins are molecules
that bind both to
bacteria and phagocytes
Opsonization increases
phagocytosis by 1,000 fold.
4. Cytolysis (MAC)
• Disrupt the membrane & the entry of water and
electrolytes into the cell

5. Enhancement of antibody production

• Binding of C3b to its receptors on the surface of
activated B cells  enhanced antibody production
Regulation of Complement System
1. C1 inhibitor
• Important regulator of classic pathway
• A serine protease inhibitor (serpin)
• Irreversibly binds to and inactivates C1r and
C1s, as well as MASP in lectin pathway

2. Factor H
• Regulate alternative pathway
• Reduce amount of C5 convertase available
• With both cofactor activity for the factor I-
mediated C3b cleavage, and decay accelerating
activity against C3bBb (C3 convertase)
3. Properdin
• Protects C3b and stabilizes C3 convertase

4. Factor I
• Cleaves cell-bound or fluid phase C3b and C4b
 inactivates C3b and C4b

5. Decay accelerating factor (DAF)

• Glycoprotein on surface of human cells
• Prevents assembly of C3bBb or accelerates
disassembly of preformed convertase 
no formation of MAC
• Acts on both classical and alternative
6. C4b-binding protein (C4BP)
• Inhibits the action of C4b in classical pathway
• Splits C4 convertase and is a cofactor for factor I

7. Complement Receptor 1 (CR-1)

• Co-factor for factor I, together with CD46

8. Protectin (CD59) and Vitronectin (S protein)

• Inhibits formation of MAC by binding C5b678
• Present on “self” cells to prevent complement
from damaging them