The Complement

System
Paper II
Unit 4.2
The complement system
• This is a part of the immune system that helps or
“complements” the ability of antibodies and phagocytic cells
by amplifying this ability, to clear pathogens from an
organism.
• It is part of the innate immune system that is not adaptable
and does not change over the course of an individual's
lifetime.
• However, it can be recruited and brought into action by
the adaptive immune system.
• The complement system consists of a number of small
proteins found in the blood, generally synthesized by
the liver, and normally circulating as inactive precursors (pro-
proteins).
• It is a complex system of about twenty serum proteins, which
interact in a cascade and activate each other sequentially.
• When stimulated by one of several triggers, proteases cleave
specific proteins to release cytokines and initiate an
amplifying cascade of further cleavages.
cleavages
• This cascade involves proteolysis of the proteins.
• The end-result of this activation cascade is massive
amplification of the response and activation of the cell-
killing membrane attack complex on the surfaces of
microorganisms.
• In addition, they aid in the defence response by dilating
blood vessels, attracting other cells of the immune system to
the site of infection, and enhancing the ability of phagocytes
to bind, ingest and destroy the microorganism.
• The complement system thus forms the major means by
which the immune system of vertebrates protects them from
bacterial infections.
• Individual deficient in complements also show immune
complex diseases, similar to those deficient in antibodies
themselves, which involve precipitation of antigen-antibody
complexes in tissues and blood vessels, causing
inflammation.
• Proteins and protein fragments that make up the
complement system are called its components, and
include serum proteins, serosal proteins, and cell membrane
receptors.
• These are synthesized mainly by the liver, and circulate in
blood and extracellular fluid.
• These proteins account for about 5% of the globulin fraction
of blood serum.
• The major proteins of the complement system are
designated as C1 to C9, factors B, D, H, I and P, while other
substances may play a regulatory role.
• The complement cascade involves triggering of the
complement components either directly through infection by
a microorganism or indirectly through the presence of an
antigen-antibody complex.
• The triggering of such a cascade involves proteolytic cleavage
of the first complement, and one of the fragments formed as
a result of lysis cleaves the next component.
• At each step, membrane binding sites are exposed, and the
subcomponents formed by cleavage bind sequentially to the
membrane of the microbe.
• This generates a cascade, ultimately leading to membrane
attack complexes on the microbial surface, bringing about its
lysis.
• The complement system works on two pathways to achieve
this.
• One is the classical pathway, which is antibody dependent,
and is activated by binding of antibodies to a foreign particle.
• The other is the alternative pathway, which is triggered by
the polysaccharides in the microbial membrane.
The Classical Complement Pathway
• This pathway is antibody dependent, and is triggered when
antibodies, specifically IgM, IgG1, IgG2 and IgG3 bind to a
foreign particle.
• The constant regions of these antibodies bind the first
component of the classical pathway, C1.
• C1 is made up of three subunits, C1q, C1r and C1s.
• The C1q subcomponent is a large protein made up of 6
identical subunits, each of which is composed of three
different polypeptide chains.
• The carboxyl terminals of these chains are folded into
independent globular structures, whereas their amino
terminals are wound together into triple helical chains, so
that the subcomponent resembles a bunch of flowers.
• C1q binds with the Ig on two adjacent domains.
• This binding attracts any of several activator molecules from
the surroundings, bringing about conformational changes
followed by proteolytic cleavage of C1r, which in turn
activates C1s.
• The activated C1s is now able to catalyze the formation of
enzyme C3 convertase from C2 2 and C4.
• C3 Convertase is actually made up of the fragment C4b of
components C4 and C2a of component C2.
• C4 is highly sensitive to activated C1s, and upon reaction,
breaks into subcomponents C4 4a and C4b.
• The C4a fragment is lost, while C4b binds with binding sites
on the antigens nearby.
• In absence of binding sites, C4
4 is not activated.
• The C4b site attracts C2 in presence of Mg++, which is also a
substrate for activated C1s.
• C2 bound to C4b cleaves into C2a
C and C2b in the presence of
activated C1s.
• The C2b fragment is lost while C2a firmly binds to C4b,
forming the C3 convertase, C4 4b2a.
• The C3 convertase is capable of enzymatically cleaving its
substrate C3, which is the most important component of the
complement system.
• This is because both the classical and the alternative
pathways ultimately converge on the lysis of C3, to proceed
towards formation of membrane attack complexes.
• C3 convertase is highly unstable, and dissociates with
temperature, or a time of just a few minutes, releasing the
C2a, if sufficient C3 is not available.
available
• On binding, C3 is cleaved into C3a and C3b.
• The former is released, and plays an important role in other
skin inflammatory reactions such as psoriasis.
• The larger fragment, C3b, forms a complex with the C3
convertase, C4b2a3b, to form the next link in the cascade C5
convertase.
• Component C5 binds with either C4b or C3b and cleaves into
fragments C5a and C5b.
• Of these, C5b binds to the surface of the target cell and
initiates the formation of membrane attack complexes with
the help of late-acting components of the complement
system.
The MBL Complement Pathway
• A protein, known as the Mannose Binding Lectin or Mannan
Binding Lectin (MBL), has the capacity to initiate the classical
complement pathway without the participation of
antibodies.
• The protein is able to bind with the antigenic epitopes,
similar to an antibody.
• MBL interacts on such binding with two other proteins,
which are MBL Associated Serine Proteases MASP and
MASP2.
• These are analogous to C1r and C1s.
• The formation of complex between MBL, MASP and MASP2
is analogous with the C1qrs complex.
complex
• The MBL complex is able to cleave the component C4.
• After this, steps similar to those in the classical pathway are
followed, leading first to the formation of C3 convertase,
then C5 convertase and finally the membrane attack
complex.
The Alternative Complement Pathway
• The alternative complement pathway constitutes the
humoral component of natural defence against antibody
independent infections.
• Six proteins, viz. C3, B,D, H,I and P recognize, initiate and
amplify the pathway to form the activator bound C3/C5
convertase.
• These activators include a variety of substances, mainly
bacterial polysachharides, as also those of fungi, viruses,
yeast, or of other animal cells, and aggregates of
immunoglobulins such as IgA and IgG.
• The fragments of the C3 component are linked through a
thiol-ester bond.
• This bond undergoes spontaneous but slow hydrolysis into
an intermediate form, C3i.
• The component B binds C3 3i, forming C3iB. Presence of
component D and Mg++ results in cleavage of B into
fragments Ba and Bb.
• Ba is released, and Bb binds with C3i, to form C3iBbMg,
known as the initial C3 convertase.
convertase
• This formation is under positive regulation of component P,
which is a cyclic protein known as Properdin.
• The components H and I, on the other hand, negatively
regulate the reaction by inactivation of the enzyme.
• The initial C3 convertase activates binding of B with C3,
resulting in cleavage of C3 into C3a and C3b.
• C3a fragment is lost, while C3bb becomes membrane bound.
• B is cleaved into Ba and Bb by D, to form C3bBb, the C3
convertase.
• The C3 convertase so formed, in presence of additional C3b,
acts as a C5 convertase, C3bBbC
bBbC3b.
The membrane Attack Complex
• This is the culmination of the complement pathways by
activation of the C5 component.
component
• The complex is a killer molecule that successfully injures the
microbial cell on the surface of which it is formed.
• It is formed by the assembly of about 20 proteins, and has a
molecular weight of approximately 1.7 million.
• It contains one molecule each of components C5b, C6, C7
and C8, along with one or more molecules of C9.
• On cleavage of C5 by the C5 convertase that is produced by
the complement pathways, a nascent C5b* is produced.
• This, along with C6, forms a bimolecular complex that binds
to C7.
• The C5b-7 so formed can insert itself into a target lipid
bilayer in close proximity, using its hydrophobic regions.
• C5b-7 thus initiates assembly of the MAC (membrane attack
Complex), and acts as a receptor for component C8.
• Binding of C8 to the C5b-7 7 causes formation of trans-
membrane channels wide enough to disturb the membrane
architecture.
• The C5b-8 attracts many C9 molecules, and their binding to
the C5b-8 facilitates further penetration of the MAC to the
hydrocarbon core of the membrane.
membrane
• The C9 molecule is tubular, and the number of C9 molecules
determine the diameter of the channel formed in the
membrane.
• In addition, C9 also attracts C9
9RP (C9 Related Protein), which
is cytotoxic, and has pore forming activity.
• The complex of C6, C7, C8, C9 9 and C9RP together is termed
as pore-forming protein or Perforin.
Perforin
• The pores formed cause the cellular content of the target cell
to leak out, eventually leading to its lysis.
Membrane Attack Complex
 Implications of the
Complement pathways in the
Immune System
Efficiency of the complement system
• The cascade of complement components is a rapidly
progressing process.
• It ensures that the microorganism is dealt with without much
delay.
• Either of the complement pathways can get activated due to
a microbe, either in presence or in the absence of
antibodies.
• Formation of membrane attack complexes provides a certain
method of destruction of the microbes.
microbes
Protection of non-target
target cells
• A protein known as the S protein inhibits MAC formation by
binding to the binding sites on C5b-7, which, though may be
able to bind with C8 and C9 9, is unable to bind with the
membrane lipids.
• The S protein protects cells adjacent to the target cell from
lysis in this manner.
Indirect role in the immune response
• The complement pathways, in addition to forming
membrane attack complexes that destroy target cells, also
release a variety of complement fragments including C3a,
C4a and C5a, together known as anaphylotoxins.
• These are low molecular weight, biologically active peptides
that act on small blood vessels, smooth muscles, mast cells
and peripheral blood leucocytes.
leucocytes