Complement System
Research on complement began in the 1890s, when Jules Bordet
showed that sheep antiserum to the bacterium Vibrio cholerae
caused lysis of the bacteria and that heating the antiserum
destroyed its bacteriolytic activity. Surprisingly, the ability to lyse
the bacteria was restored to the heated serum by adding fresh
serum that contained no antibodies directed against the
bacterium and was unable to kill the bacterium by itself.
Bordet reasoned that bacteriolytic activity requires two different
substances: first, the specific antibacterial antibodies, which
survive the heating process, and a second, heat-sensitive
component responsible for the lytic activity.
Paul Ehrlich in Berlin independently carried out similar
experiments and coined the term complement, defining it as “the
activity of blood serum that completes the action of antibody.”
After initial activation, the various complement
components interact, in a highly regulated cascade, to carry
out a number of basic functions including:
• Lysis of cells, bacteria, and viruses.
• Opsonization, which promotes phagocytosis of
particulate antigens.
• Binding to specific complement receptors on cells of the
immune system, triggering inflammation.
• Immune clearance, which removes immune complexes
from the circulation and deposits them in the spleen and
liver.
The proteins and glycoproteins that compose the complement
system are synthesized mainly by liver hepatocytes, although
significant amounts are also produced by blood monocytes,
tissue macrophages, and epithelial cells of the gastrointestinal
and genitourinary tracts.
These components constitute 5% (by weight) of the serum
globulin fraction. Most circulate in the serum in functionally
inactive forms as proenzymes or zymogens, which are inactive
until proteolytic cleavage, which removes an inhibitory
fragment and exposes the active site. The
complement-reaction sequence starts with an enzyme
cascade.
1. Classical pathway
2. Alternative pathway
Classical pathway
Alternative pathway
In the classical pathway, C3 is rapidly cleaved to C3a and C3b by the
enzymatic activity of the C3 convertase. In the alternative pathway,
serum C3 is subject to slow spontaneous hydrolysis to yield C3a and
C3b.
The C3b component can bind to foreign surface antigens (such as
those on bacterial cells or viral particles) or even to the host’s own
cells.
The membranes of most mammalian cells have high levels of sialic
acid, which contributes to the rapid inactivation of bound C3b
molecules on host cells; consequently this binding rarely leads to
further reactions on the host cell membrane.
Because many foreign antigenic surfaces (e.g., bacterial cell walls,
yeast cell walls, and certain viral envelopes) have only low levels of
sialic acid, C3b bound to these surfaces remains active for a longer
time.
Lectin pathway
Lectins are proteins that recognize and bind to specific
carbohydrate targets. (Because the lectin that activates
complement binds to mannose residues, this is called as the
mannan-binding lectin pathway.)
The lectin pathway, like the alternative pathway, does not depend
on antibody for its activation. However, the mechanism is more
like that of the classical pathway, because after initiation, it
proceeds, through the action of C4 and C2, to produce a C5
convertase.
The lectin pathway is activated by the binding of mannose-binding
lectin (MBL) to mannose residues on glycoproteins or
carbohydrates on the surface of microorganisms including certain
Salmonella, Candida albicans, etc.
MBL is an acute phase protein produced in
inflammatory responses. Its function in the
complement pathway is similar to that of C1q, which
it resembles in structure.
After MBL binds to the surface of a cell or pathogen,
MBL-associated serine proteases,MASP-1 and
MASP-2, bind to MBL.
The active complex formed by this association
causes cleavage and activation of C4 and C2. The
MASP-1 and -2 proteins have structural similarity to
C1r and C1s and mimic their activities.
Membrane-Attack Complex (MAC)
The terminal sequence of complement activation involves C5b, C6,
C7, C8, and C9, which interact sequentially to form a
macromolecular structure called the membrane-attack complex
(MAC). This complex forms a large channel through the membrane
of the target cell, enabling ions and small molecules to diffuse
freely across the membrane.
The end result of activating the classical, alternative, or lectin
pathways is production of an active C5 convertase. The C5b
component is extremely labile and becomes inactive within 2
minutes unless C6 binds to it and stabilizes its activity.
Up to this point, all the complement reactions take place on the
hydrophilic surface of membranes or on immune complexes in the
fluid phase.
As C5b6 binds to C7, the resulting complex undergoes a
hydrophilic-amphiphilic structural transition that exposes
hydrophobic regions, which serve as binding sites for membrane
phospholipids.
If the reaction occurs on a target-cell membrane, the
hydrophobic binding sites enable the C5b67 complex to insert
into the phospholipid bilayer.
Binding of C8 to membrane-bound C5b67 induces a
conformational change in C8, so that it too undergoes a
hydrophilic-amphiphilic structural transition, exposing a
hydrophobic region, which interacts with the plasma
membrane. The C5b678 complex creates a small pore, 10 Å in
diameter; formation of this pore can lead to lysis of RBC but not
of nucleated cells.
The final step in formation of the MAC is the binding and
polymerization of C9, a perforin-like molecule, to the
C5b678 complex. As many as 10–17 molecules of C9 can be
bound and polymerized by a single C5b678 complex. During
polymerization, the C9 molecules undergo a
hydrophilic-amphiphilic transition, so that they too can
insert into the membrane.
The completed MAC, which has a tubular form and
functional pore size of 70–100 Å, consists of a C5b678
complex surrounded by a poly-C9 complex. Since ions and
small molecules can diffuse freely through the central
channel of the MAC, the cell cannot maintain its osmotic
stability and is killed by an influx of water and loss of
electrolytes.
The smaller fragments resulting
from complement cleavage, C3a,
C4a, and C5a, called
anaphylatoxins, bind to receptors
on mast cells and basophils and
induce degranulation, with release
of histamine and other
pharmacologically active
mediators.