Complement System
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Complement System
The complement system constitutes a cascade of interactions between a large number of
heat-labile plasma proteins called complements. These proteins are inactively circulating in
the plasma and are functional when cleaved into peptide fragments. The activation of the
complement system occurs through three different pathways: the classical pathway, the
mannose-binding lectin (MBL) pathway, and the alternative pathway.
Activation of complement involves the sequential proteolysis of proteins to generate enzyme
complexes with proteolytic activity. Proteolytic cascades allow tremendous amplification
because each enzyme molecule activated at one step can generate multiple activated
enzyme molecules at the next step. The products of complement activation become
covalently attached to microbial cell surfaces, to antibodies bound to microbes and to other
antigens, and to apoptotic bodies. Complement activation is inhibited by regulatory proteins
that are present on normal host cells and absent from microbes. The regulatory proteins are
an adaptation of normal cells that minimize complement-mediated damage to host cells.
Microbes lack these regulatory proteins, which allows complement activation to occur on
microbial surfaces.
Complement activation takes place through any of the following three
pathways:
The classical pathway
The alternative pathway
The lectin pathway
What is the classical pathway of the
complement system?
The classical pathway of the complement system is
activated by the formation of antigen-antibody
complexes (immune complexes). Other activators
include apoptotic cells, C-reactive protein, bacterial
polysaccharides, and nucleic acids, which trigger
classical pathways in antibody independent manner.
The complement proteins in this pathway are
named numerically from 1 to 9 (C1 to C9), each with
subunits of their own. All the larger subunits of
these proteins are designated as “b” and smaller
subunits as “a” while the reverse case occurs in the
case of protein C2 (C2a is a larger fragment and
Steps/mechanism/process of classical
pathway of the complement system
1. The formation of antigen-antibody complex induces a conformational
change in the Fc portion of the antibody, exposing a binding site for
the C1 component of complement. C1 protein is a macromolecular
complex with the following subunits: 1 C1q, 2 C1r, 2 C1s
2. C1q has six subunits, each composed of three homologous chains
(A, B, and C) with extended tails forming six globular heads. Each
globular head acts as a binding site for antibody Fc. C1q globular
heads must bind at least two Fc sites of the antibody to maintain
stable C1-antibody interaction. Antigen-bound IgM undergoes
conformational change exposing at least three binding sites for C1q
whereas IgG has only one binding site for C1q.
3. As C1q binds to antibodies through its globular heads, conformational changes occur in the C1r
proenzyme. The activated C1r now cleaves C1s activating C1s serine protease.
4. Activated C1s tend to activate another component, C4, by cleavage. This bound C1s cleaves C4
forming C4b, which binds covalently to the bacterial surface membrane.
C4b binds C2, thereby exposing it to cleavage by C1s, forming the C4b2a enzyme
complex. This enzyme complex is called C3 convertase and catalyzes the central
reaction of the complement pathway. This enzyme deposits a large number of C3b
molecules on the surface of the pathogen.
This membrane-bound enzyme complex C4b2a3b is called C5 convertase. This enzyme
in turn, cleaves C5 into C5a and C5b.
C5b then triggers the formation of membrane attack complex (MAC), i.e., C5b-9. C5b
attaches to C6, followed by C6 and C7 forming complex C5b-6-7, and this complex acts
as a receptor for C8 and C9. After polymerization, finally, the MAC complex is formed.
Inhibitors of the classical pathway
The complement system must be controlled to prevent unwanted
host cell attacks. Inhibitors present in host tissues are present in the
soluble or membrane-bound form. Decay acceleration activity (DAA)
and factor I cofactor activity (CA) are the two main regulation
mechanisms that accelerate the rate of dissociation of C3 convertase
and factor I-mediated cleavage of C3b-C4b incapable of reforming the
C3 convertase, respectively.
Some of the complement inhibitors
include:
1. Soluble regulators:
C1-Inhibitor (C1-INH): C1 is inhibited by plasma serine proteinase
belonging to the serpin superfamily, also known as C1 esterase
inhibitor.
Factor I: It cleaves C3b and C4b, thus blocking the formation of C3
and C5 convertase.
Others: C4bp, carboxypeptidase N, Factor H
2. Membrane-bound proteins:
CD35 (Complement receptor 1 or CR1): In humans, it is present on antigen-presenting cells,
erythrocytes, etc. It accelerates the decay of the C3/C5 convertase. It also acts as a co-factor for
factor I in the degradation of C3b and C4b.
CD46 (Membrane co-factor protein or MCP): It acts as a co-factor for factor I in the degradation of
C3b and C4b.
CD55 (Decay accelerating factor or DAF): It inhibits and accelerates the decay of C3 convertase.
CD59 (protectin): It inhibits the pathway at the terminal step by inhibiting the insertion of the C9 to
C5b-8 complex hence preventing MAC assembly.
Others: CrrY
Significance/Applications of classical
pathway of the complement system
This pathway plays a key role in the opsonization and removal of nuclear debris. Bacteria and viruses
are easily phagocytosed in the presence of C3b as C3b receptors are numerous on the surface of
phagocytes.
The small fragments C4a,3a, and C5a are important mediators of inflammation, while C2a is the
precursor of vasoactive kinin.
Membrane attack complex forms a pore or transmembrane channel, allowing free exchange of ions
between cell and the external surrounding. The rapid influx of ions into the cell increases osmotic
pressure inside the cell, which eventually bursts.
Autoantibodies can also trigger this pathway leading to diseases like hemolytic anemia, myasthenia
gravis, and bullous pemphigoid.
The alternative pathway
It was discovered later but is phylogenetically older than the classical pathway, is triggered when
a complement protein called C3 directly recognizes certain microbial surface structures, such as
bacterial LPS. C3 is also constitutively activated in solution at a low level and binds to cell
surfaces, but it is then inhibited by regulatory molecules present on mammalian cells. Because
microbes lack these regulatory proteins, the spontaneous activation can be amplified on
microbial surfaces. Thus, this pathway can distinguish normal self from foreign microbes on the
basis of the presence or absence of the regulatory proteins.
The alternative pathway (AP) is dominant to the classical and lectin pathway during normal
physiological conditions. This pathway is constantly activated at a low level in which C3 gets
hydrolyzed to C3b. If foreign biological and artificial surfaces like carbohydrates, lipids, proteins,
gas bubbles, etc., are encountered, this pathway gets automatically triggered. The continuous
activation of C3 is termed “tick over” which generates low levels of C3b and gets amplified if
needed.
Steps / Mechanism / Process of Alternative pathway of
the complement system
The components of this pathway include:
C3, C5-C9
Factor B
Factor D
Factor P (properdin)
Upon encountering foreign surfaces, the 3b protein binds directly to these surfaces and, unlike the
classical pathway, does not require the formation of antigen-antibody complexes.
The hydrolyzed thioester domain of C3 undergoes structural change exposing the binding site for
Factor B. Mg2+ is the only ion required for functional activation of the alternative pathway that
stabilizes the interaction between C3b and factor B.
This C3(H2O) bound Factor B is cleaved by serine protease Factor D, forming C 3(H2O)Bb, C3
convertase. C3 convertase in the alternative pathway is unique as it is made up of subunits C3b and
Bb. This complex can now activate more C3 via a powerful amplification cycle; therefore, this
pathway is also called “the amplification loop”.
The convertase complex, in turn, cleaves C3 to C3a and C3b. The 3a fragment is released into the
fluid phase.
When a second C3b adds to it, the C5 convertase is synthesized.
The membrane attack complex is formed following the same pathway as the classical pathway of
the complement system. C5 convertase cleaves C5 into subunits C5a and C5b. C5b recruits C6, C7,
C8, and C9 forming MAC that induces cell lysis.
Three distinct pathways to initiate alternative
pathway has been discovered:
The alternative tickover pathway
The above-mentioned pathway that constitutes spontaneous hydrolysis
of C3 to C3(H2O) is termed the tickover pathway.
The alternative properdin-activated pathway
The serum protein properdin promotes the synthesis of AP convertases.
It is synthesized by monocytes, granulocytic cells, and T cells. The C3Bb
complex has a half-life of only 90 seconds under physiological conditions
but properdin, when gets associated with this complex, makes it more
stable.
Besides maintaining the stability of convertases, properdin may also
initiate an alternative pathway. Properdin, when bound to components of
microbial membranes in the presence of Mg 2+, can now bind C3b and
factor B.
The alternative protease-activated pathway
Proteases of blood coagulation pathways such as thrombin and
plasmin could cleave C3 and C5, releasing C3a and C5a, respectively.
This process can enhance the activation of complement cascades.
Regulators of the Alternative Pathway
As this pathway does not require any immune response and is
activated independently upon encountering foreign particles, it must
be rigorously controlled to prevent unwanted host cell attacks.
Complement factor H (CFH): It is a soluble inhibitor of C3 convertase. It
acts as a cofactor in the cleavage of C3b to iC3b. This factor is required
to control physiologic, spontaneous hydrolysis of the C3 molecule
under control.
CFH inhibits the formation of the C3 convertase by competing with
factor B. It accelerates the dissociation of the C3bBb convertase
complex. The deficiency of this factor leads to kidney abnormalities
such as atypical hemolytic uremic syndrome (aHUS).
Membrane cofactor protein (MCP), decay acceleration factor (DAF), and complement
receptor 1 (CR1) are membrane inhibitors of C3 convertase. MCP and CR1 are cofactors
for the inactivation of C3b and C4b that occurs via complement factor I (CFI).
CFI is a plasma protease that prevents the reassembly of C3 convertase by proteolytic
inactivation of C3b to iC3b (inactive C3b).
Complement factor H-related protein (CFHR), P-selectin and heme are activators of C3
convertase. CFHR has a control function by competing with CFH for binding to C3b.
Applications / Significance of Alternative pathway
This pathway monitors pathogen invasion in normal physiological
conditions.
Infiltrating cells such as neutrophils bring in C3 and properdin that
can initiate alternative pathways.
Factor B is an acute phase protein and increases during inflammation.
C5a and C3a are important mediators of inflammation.
Cell lysis and inflammation are induced by MAC.
Lectin Pathway of the complement system
The lectin pathway (LP) proceeds like the classical pathway by
activating the C3 convertase. The binding of mannose-binding
proteins initiates the cascade of reactions in this pathway in blood
plasma called lectins to mannose-containing proteoglycans on the
surface of pathogens. Pattern recognition molecules of LP other than
lectins are collectins, ficolins H, L, and M.
Definition and Introduction
The lectin pathway of the complement system in the innate immune
system includes a cascade of enzymatic reactions in which
carbohydrate-binding proteins called lectins initiate the reaction after
binding to the surface of a wide range of pathogens. The pathway is
based on the activation of mannose-binding lectin-associated serine
proteases (MASPs), which are soluble serine proteases of serum.
Steps, Mechanism, Process of Lectin Pathway
Mannose Binding Lectin (MBL) is structurally similar to C1q (has a
collagen-like region and globular heads) in the classical pathway and
allows binding on many pathogens. The recognition of mannose-
containing sugars on pathogens by MBL initiates a reaction. Like C1q
in the classical pathway, MBL is a six-headed molecule that forms a
complex with 2 protease zymogens MASP-1 and MASP-2, homologous
to C1r and C1s.
Except for MBL; ficolin 1, 2, and 3 are also used as pattern recognition
receptors.
When the MBL complex binds to the pathogen’s surface, MBL-associated serine proteases MASP-1
and MASP-2 get activated, which cleave C4 and C2. MBL/ MASP-1/ MASP-2 form a trimolecular
complex that activates MASPs and results in subsequent cleavage of C4.
In physiological conditions, MASP-1 is required for the cleavage of MASP-2, and both activated proteases
can cleave C2, whereas MASP-2 can also cleave C4. MASP-1,2 and 3 are synthesized as proenzymes.
C4 is cleaved into C4a and C4b. C4b attaches to the surface of the pathogens inducing C2 to bind,
which is cleaved by MASP2 to form C2b and C2a. C2a binds to C4b, forming complex C3 convertase.
The convertase complex, in turn, cleaves C3 to C3a and C3b. The 3a fragment is released into the
fluid phase.
This complex C4b2a3b, the C5 convertase, synthesis proceeds on the surface membrane of the
pathogen.
The membrane attack complex (MAC) is formed following the same pathway as the classical pathway
of the complement system. C5 convertase cleaves C5 into subunits C5a and C5b. C5b recruits C6,
C7, C8, and C9 forming MAC that induces cell lysis.
Inhibitors of Lectin Pathway
Like any other pathway of complement activation, uncontrolled activation of the lectin
pathway can lead to serious damage.
Some inhibitors include C1-inhibitor, antithrombin (AT), and α(2)-macroglobulin that
inhibits MASP-1 and MASP-2. Another important component Map44 (Mannose-binding
lectin-associated protein of 44 Kilodalton), has been suggested to act as a competitive
inhibitor of lectin pathway activation by displacing MASP-2 from MBL, hence preventing
cleavage of C4 and C2.
Applications and Significance of Lectin Pathway
This pathway effectively protects against invading pathogens and
apoptotic cells. Example: In dengue, this pathway has been found to
prevent virus attachment to the target cells.
Activation of the pathway induces:
inflammatory reactions (C5a, C4a, and C3a are important mediators
of inflammation),
opsonization (C3b, C4b, C1q)
phagocytosis and lysis of target pathogens (via Membrane attack
complex)