COMPONENT COMPLEMENT

SYSTEM

Lecturer
Mr.Justine Malemba
 The complement system
One of the major effectors of innate immunity.

It is also an effector of humoral immunity.

Derived from experiments performed by Jules

Bordet shortly after the discovery of antibodies.

 The complement system
He demonstrated that if fresh serum containing an

antibacterial antibodies was added to bacteria at

37 degrees , the bacteria were lysed.
However, if the serum was heated at 56 degrees

or more, it lost its lytic capacity.

 The complement system
The loss was not due to decay of antibodies

activity because antibodies are heat stable.

He concluded that the serum contain another

heat –labile component that assists or

complements the lytic function of antibodies.
This component was later given the name

complement.
 The complement system
Complement system consists of serum and cell surface

proteins.
These proteins interact with one another in regulated

manner to generate products that function to eliminate

microbes.
Thus; by definition, complement are plasma proteins

that are normally inactive and only activated upon

binding to microbes or antibodies.

The complement system

Activation of complements involves the sequential

proteolysis of proteins to generate newly

assembled enzyme complexes with proteolytic
activity.
 The complement cascade.
The complement cascade may be activated by one

of the three pathways.

 The alternative pathway.

 The classical pathway.

 The lectin pathway.

The complement cascade.
The pathways of complement activation differ in

how they are initiated.

However, all of them result in the generation of

enzyme complexes that cleave the most abundant

complement protein C3.
Activation of pathways.

 Alternative pathway.

Activated on the microbial cell surface in the

absence of antibodies.
 Classic pathway.

Activated when antibodies bind to microbes or

other antigens(Ags).
Activation of pathways
 Lectin pathway.

Activated when a plasma protein, mannose -

binding lectin binds to terminal mannose residues.

The alternative pathway.
C3 is the most abundant complement protein.

Activation results in the proteolysis of C3.

Proteolysis of C3 generates C3a and C3b.

C3b attaches to microbial surface and C3a is

released.
Plasma protein called factor B binds to C3b which

is bound to microbial surface.

The alternative pathway
Bound factor B is in turn cleaved by factor D

releasing;
 A small fragment called Ba.

 A larger fragment called Bb.

Bb remains attached to C3b forming C3bBb.

 Another protein called Properdin binds to

stabilize the C3bBb.

The alternative pathway
C3bBb complex is known as C3 convertase which

cleaves more C3 to yield more C3b and C3a.

C3b binds to more bacterial cell walls.

Some of the C3b generated by C3 convertase bind

to the C3 convertase itself forming C3bBbC3b.

C3bBbC3b is known as C5 convertase.

C5 convertase cleaves C5 into C5a and C5b.

The alternative pathway
C5a is released .

C5b binds and activate C6,C7,C8 and C9 to form

cylindrical cytocidal membrane attack

complex(MAC).
This membrane attack causes lysis of bacteria.
Alternate Pathway
The classical pathway

Initiated by binding of complement protein C1 to

the IgG or IgM antibody that have bound antigen.

C1 is a large protein complex composed of;

 C1q,C1r and C1s subunits.

• C1q binds to the antibody and C1r and C1s are

proteases.
The classical pathway
Antibodies have binding sites for C1q.

However, these binding sites are only accessible

when an antibody binds to an antigen causing

conformation change of C1q.
Binding of C1q to the Fc region of IgM or IgG

activates C1r.
The classical pathway
Activated C1r cleaves and activates C1s.

Activated C1s cleaves the next protein in the

cascade,C4 generating C4a and C4b.

Note:

 C4 is homologous to C3 in alternative pathway.

 C4b contains thioester bonds similar to that in C3b.

The classical pathway
The smaller C4a fragment is released in the fluid

environment.
C4b attaches to antibody -antigen complex.

C2 then complexes with the cell surface- bound

C4b.
Complexed C2 is cleaved by a near by C1s yielding

C2a and C2b.

The classical pathway

C2a is released (has unknown biological activity).

Larger C2b fragment remains physically associated

with C4b on the cell surface.

The resulting C4b2b complex is the classical

pathway C3 convertase.
The classical pathway
This C3 convertase has the ability to bind and

cleave C3.
Cleavage of C3 generates C3a and C3b.

C3b forms covalent bonds with cell surfaces or

with antibody where complement activation was

initiated.
The classical pathway

Once C3b is deposited, it can bind factor B and

generate more C3 convertase by the

alternative pathway.

Factor B in alternative pathway is homologous

to C2.
The classical pathway
Some of the C3b generated by classical pathway

C3 convertase bind to the convertase itself

forming C4b2b3b complex.
The classical pathway
This complex function as the classical pathway C5

convertase.
This C5 convertase cleaves C5 to generate C5a and

C5b.
The subsequent steps are similar to alternative

pathway.
Classical Pathway
The lectin pathway

 Activation is triggered in the absence of antibody by

the binding of microbial polysaccharides to

circulating lectins.
Circulating lectins include;

 Plasma mannose ( or mannan)- binding lectin (MBL).

 N-acetylglucosamine recognizing lectins known as

ficolins.
 The lectin pathway
MBL binds to mannose residues on polysaccharides

of microbes.
MBL also binds to MBL- associated serine
proteases(MASPs) such as; MASP-1,MASP-2 and
MASP-3.
MBL is typically associated with MASP-2 and MASP-

3.
The lectin pathway
These two proteases form complex similar to the

ones formed by C1r and C1s.

MASP-2 cleaves C4 and C2.

Subsequent events in this pathway are identical to

those that occur in the classical pathway.

The role of different complement
components in immunity.
Principal effector functions of the complement

system in innate immunity and specific humoral

immunity are;
 To promote phagocytosis of microbes.

 To stimulate inflammation.

 To induce lysis of microbes by (MAC).

The role of different complement
components in immunity.
In addition, products of complement facilitates

the activation of B-lymphocytes and the

production of antibodies.
The role of complement system in
host defense.
 Opsonization and phagocytosis.

Opsonization means “to make tasty” putting

butter on bread.
Microbes become coated with C3b or C4b and are

phagocytosed by the binding of these proteins to

specific receptors on macrophages and
neutrophils.
• C3b and C4b are opsonins.

 Stimulation of inflammatory responses.

C5a,C4a and C3a induce inflammation by

activating mast cells and neutrophils.
These smaller fragments(peptides) bind to mast

cells and induce degranulation.

Degranulation results in the release of vasoactive

mediators such as histamine.

In neutrophils,C5a stimulates

 Motility.

 Firm adhesion to endothelial cells.

 Respiratory burst and production of reactive

oxygen intermediates.
The role of complement system in
host defense
C5a may act directly on vascular endothelial cells

and induce increased vascular permeability and

expression of p- selectin which promotes
neutrophil binding.
C5a is the most important mediator of mast cell

degranulation
The role of complement system in
host defense
 Complement- mediated cytolysis.

Is mediated by the MAC.

Unfortunately, Most pathogens have evolved thick

cell walls or capsules that impede access of MAC to

their cell membranes(evasion strategy).
MAC works well on organisms with thin walls such as

those of genus Neisseria.

The role of complement system in
host defense
Genetic defect in MAC components results in

increased susceptibility to infections caused by

such organisms.
 Receptors for complement proteins
Complements exert their biological activities by

binding to membrane receptors expressed on various

cell types.

 Type 1 complement receptor(CR1,or CD35).

CR1 is a high- affinity receptor for C3b and C4b.

Promotes phagocytosis of C3b and C4b- coated

particles.
Receptors for complement
proteins
It is expressed on blood cells such as;

 erythrocytes,

 neutrophils,

 monocytes,

 Eosinophils.

 T and B lymphocytes
Receptors for complement proteins

 Type 2 complement receptor(CR2,or CD21).

 stimulates humoral immune responses by

enhancing B cell activation.
CR2 is present on

 B lymphocytes

 Dendritic cells

 Epithelial cells.
Receptors for complement
proteins
 In humans, CR2 is the cell surface receptor for

Epstein – Barr virus.

 Type 3 complement receptor(MAC-1
(CR3,CD11bCD18).
Receptor for iC3b fragments generated by
proteolysis of C3b.
Receptors for complement proteins
Mac-1 is expressed on the following cells;

 Neutrophils

 Mast cells

 NK cells

Promotes phagocytosis of microbes opsonized by

C3b.
May direct and recognize bacteria for
phagocytosis by binding to microbial molecules.
Receptors for complement
proteins
 Type 4 complement receptor(CR4,CD11cCD18).

 Its functions are similar to CR3.

Regulation of complement activation
Complement activation cascade is highly
regulated.
Regulation is mediated by cell and membrane

proteins.
Regulation is needed for two reasons;

 Spontaneous and continuous generation of

complements can damage normal cells and
tissues.
Receptors for complement
proteins
Different regulatory mechanisms function by:

(a) Inhibiting the formation of C3 convertase.

(b) Breaking down and inactivating C3 and C5

convertases.
Regulation of complement activation

(c) Inhibiting the formation of MAC.

proteolytic activity of C1r and C1s is inhibited by

plasma protein called C1 inhibitor (C1 INH).

MAC formation is inhibited by membrane protein

called CD59.
Evasion of complements by microbes

Pathogens have evolved diverse mechanisms for

evading the complement system.

Some of the Mechanisms employed by microbes

include;
 Expressing thick walls to avoid MAC e.g. gram +ve

bacteria and fungi.

 Recruiting host complement regulatory proteins.
Evasion of complements by microbes
 Expressing sialic acid which can inhibit alternative

pathway by recruiting factor H which displaces

C3b from Bb.
 Scavenging sialic acid from the host and
enzymatically transfer the sugar to their cell
surfaces.
 Synthesizing proteins which are antagonist of the

C5a.(Staph spp).
 Summary
Complement system plays a major role in both

innate and adaptive humoral immunity.

Three ways in which complement system is

activated.

 Alternative pathway- no Ab involved(innate

immunity).

 Classical pathway–involves Ab(adaptive hummoral

immunity).
Summary
All pathways have common final steps of activation.

Complements have various biological activities.

They are tightly regulated.

Organisms evade complement system by;

 Expressing thick walls to avoid MAC

 Recruiting regulatory protein

 Producing proteins antagonist to C5a

• Their deficiencies lead to multiple infections