THE COMPLEMENT SYSTEM

GSP 2620 General and systemic pathology

Dr Simpokolwe K
Objectives
• Define complement
• Describe importance/role of complement in
immune system
• Describe the different pathways of
complement
• Describe the functions of complement
• The complement system is a part of the immune
system that enhances (complements) the ability
of antibodies and phagocytic cells to
clear microbes and damaged cells from an
organism, promote inflammation, and attack the
pathogen's cell membrane.
• It is part of the innate immune system, which
is not adaptable and does not change during
an individual's lifetime. The complement
system can, however, be recruited and
brought into action by antibodies generated
by the adaptive immune system.
• The complement system consists of a number of
small proteins that are synthesized by the liver,
and circulate in the blood as inactive precursors.
When stimulated by one of several
triggers, proteases in the system cleave specific
proteins to release cytokines and initiate an
amplifying cascade of further cleavages.
• The end result of this complement
activation or complement fixation cascade is
stimulation of phagocytes to clear foreign and
damaged material, inflammation to attract
additional phagocytes, and activation of the
cell-killing membrane attack complex.
• Over 30 proteins and protein fragments make
up the complement system, including serum
proteins, and cell membrane receptors.
• Three biochemical pathways activate the
complement system:
i. The classical complement pathway
ii. The alternative complement pathway
iii. The lectin pathway.
Components of the complement system
• There are more than 20 components of which the
following are important.
1. C1,C2,C3,C4,C5,C6,C7,C8,C9
2. Factor B,
3. Factor D,
4. Factor H,
5. Factor I,
6. Properdin,
7. C1 inhibitor,
8. C4 binding protein,
9. S protein.
THE CLASSICAL PATHWAY
• The classical pathway is phylogenetically newer
and is named “classical”, only because it was
discovered earlier. It is activated by Antigen-
Antibody complexes. The classical pathway
consists of 4 proteins C1, C2, C3 and C4.
Summary Steps in classical pathway:
• C1 is a complex made of hexamer C1q and serine
proteases C1r and C1s.
• C1q binds to Fc region of antibody. IgM is more
efficient because of it’s high avidity as it can bind
to 3 C1q compared to IgG which can bind to only
one.
• Activated C1s cleaves C4 and C4b either binds to
antigen-antibody complex or to the adjacent cell
surface.
• C1s also cleaves C2 and the C4bC2a complex is
the C3 convertase of classical pathway.
• Once C3b is formed – it can either follow
alternative pathway or bind to C4bC2a (C3
convertase) to form C5 convertase.
• Similarly, Membrane attack complex(MAC) is
formed.
• C1 binds to Fc portion of IgM or IgG near hinge-
region. This region is exposed when the antigen
binds to the antibody.
• The classical pathway begins with the formation of
antigen-antibody complex (immune complex). When
an antigen enters the body, the antibody (IgM/IgG)
binds to it. This induces conformational changes in the
Fc portion of the antibody which exposes a binding site
for C1 protein. Hence, the antibody activates the
complement system only when bound to an antigen.
• C1 is a large, multimeric, protein complex composed of
one molecule of C1q and two molecules each of C1r
and C1s subunits. C1q binds to the antigen bound
antibody (Fc portion). C1r and C1s are proteases which
help to cleave C4 and C2.
• The immune complex bound to C1 calls another
protein C4 which is cleaved into C4a and C4b. C4a goes
away whereas activated C4b attaches to the target
surface near C1q. Now, C4b attracts C2 which is also
cleaved into C2a and C2b. C2a binds C4b forming the
C4b2a complex whereas C2b goes away. The active
C4bC2a activates C3. The C4b2a complex is also known
as C3 convertase as this converts C3 into an active
form by separating C3a and C3b. One molecule of
C4b2a can cleave a large number of C3 molecules. C3b
binds to the microbial surface or to the convertase
itself.
• C3b when binds to C3 convertase forms C4bC2aC3b (C5
convertase) which activates C5.
• C5 convertase cleaves C5 into C5a and C5b. C5a
diffuses away but C5b is stabilized by binding C6. Then
C5bC6 binds to C7. C5bC6C7 complex is then inserted
into the phospholipid bilayer of the cell membrane
which further binds C8. These all (C5b678) activate C9
to form a macromolecular structure called
the membrane attack complex (MAC). This makes hole
in the bacterium, as a result, the intracellular contents
leak out and unwanted substances get in. Thus, the cell
cannot maintain its osmotic stability and the lysis
occurs by an influx of water and loss of electrolytes.
THE ALTERNATIVE PATHWAY
• It is phylogenetically more primitive pathway
compared to the classical pathway but was
named “alternative” probably because it was
discovered later among the two. This pathway
consists of proteins known by the term “Factors”
like Factor B, Factor D.
Steps in alternative pathway:
• Spontaneous hydrolysis of C3.
• C3b product is very reactive and can bind to
invader’s cell surface.
• If C3b cannot find the cell surface to bind within
60 microseconds – it is hydrolyzed.
• Factor B binds C3b on cell surface.
• Factor D cleaves Factor B, leaving “Bb” which has
a “chainsaw mechanism”
• C3bBb acts as C3 convertase which cleaves C3 –
this creates a continuous loop and many C3b can
be deposited on the invader’s cell surface.
• C3bBb together with another C3b molecule can
act as C5 convertase which cleaves C5
• C5b together with C6, C7, C8 and C9 forms MAC
• C5b, C6, C7, and C8 form a “stalk” that anchors
the complex in the bacterial cell membrane. Then
C9 proteins are added to make a channel that
opens up a hole in the surface of the bacterium.
• When a bacterium enters the host body, as a result of
inflammation, complements reach towards the site,
where C3 molecules directly touch antigen and become
active. In this pathway, serum C3 containing an
unstable thioester bond undergoes slow spontaneous
hydrolysis to yield C3a and C3b. C3b binds the surface
of foreign cell and then binds to another serum protein
called factor B. Now the factor B exposes the site which
serves as the substrate for enzymatically active serum
protein D. Then factor D cleaves B into Ba and Bb
forming C3 convertase (C3bBb). C3 convertase then
forms C5 convertase which ultimately forms a MAC as
in classical pathway.
THE LECTIN PATHWAY
• The lectin pathway consists of C2, C3, C4 and some
calcium-dependent lectin family proteins which are
homologous to C1 component.
Steps in Lectin Pathway:
• In the blood, MBL (Mannose Binding Lectin) binds to
another protein called MASP (Mannan Associated
Serine Protease).
• When MBL grabs its target (mannose on the surface of
a bacterium, for example), the MASP protein functions
like a convertase to clip C3 complement proteins to
make C3b
• Some bacteria can activate complement system
without having antibody and endotoxin. This occurs
through MBL pathway which is activated when
circulating lectin (MBL) binds to mannose residues on
glycoproteins or carbohydrates on the surface of
microorganisms. Microorganisms inducing MBL
pathway are bacteria, such as Salmonella, Listeria, and
Neisseria strains, some fungi and some viruses
including HIV-1. MBL is an acute phase protein and its
concentration increases during inflammation. The
lectin recognizes and binds the carbohydrate of the
target cell which then activates complements.
• MBL pathway resembles classical pathway as it
proceeds through the action of C4 and C2 to produce
activated proteins of the complement system. MBL
works same as C1q which it resembles in structure.
• After the MBL binds to carbohydrate residues on the
surface of a cell or pathogen, two components, MASP-
1 and MASP-2 bind to MBL. MASP stands for MBL-
associated serine proteases. Two proteases form a
tetrameric complex similar to the one formed by C1r
and C1s and cleaves C4 and C2 forming C3 convertase.
The process now continues to form of C5 convertase
and the MAC as in classical pathway.
FUNCTIONS OF COMPLEMENTS
Some major functions of complements are:
[Link] and phagocytosis
• C3b, bound to immune complex or coated on
the surface of pathogen, activate phagocytic
cells. These proteins bind to specific receptors
on the phagocytic cells to get engulfed.
[Link] lysis
• Membrane attack complex formed by C5b6789
components ruptures the microbial cell surface
which kills the cell.
[Link]
• Complement fragments attract neutrophils and
macrophages to the area where the antigen is
present. These cell surfaces have receptors for
complements, like C5a, C3a, thus, run towards
the site of inflammation, i.e. chemotaxis.
4. Activation of mast cells and basophils and
enhancement of inflammation
• The proteolytic complement fragments, C5a,
C4a, and C3a induce acute inflammation by
activating mast cells and neutrophils.
• All three peptides bind to mast cells and
induce degranulation, with the release of
vasoactive mediators such as histamine.
• These peptides are also called anaphylatoxins
because the mast cell reactions they trigger
are characteristic of anaphylaxis.
• Binding to specific complement receptors on
cells of the immune system, they trigger
specific cell functions, inflammation, and
secretion of immunoregulatory molecules.
5. Immune clearance
• The complement system removes immune
complexes from the circulation and deposits
them in the spleen and liver. Thus it acts as
anti-inflammatory function. Complement
proteins promote the solubilization of these
complexes and their clearance by phagocytes.
Summary