BT-302 Immunology: Complement System

Chapter # 11: Complement System

Introduction to Complement System

Complement system is comprised of heat labile serum proteins which are able to destroy or lyse
pathogens. This form of host defense is termed as one way of host defense against potential
pathogens. Complement proteins become inactive by heating serum at 56°C for 30 minutes. This
system is composed of more than twenty (20) proteins. These proteins are produced by variety of
body’s cells including hepatocytes (Liver cells), macrophages & gut epithelial cells. Among
complement proteins, some proteins has the ability to bind with immunoglobulin molecules
while some are involved in binding on the membrane component of various immune cells. Most
of complement proteins exist in inactive form which are also called as Proenzymes. Such
proteins need activation into active form for proper functioning. These proteins when activated
can cleave one or more other complement proteins. Such kind of activation process is called as
cascading for complement activation.

Functions of Complement Proteins

Complement system provides specific as well as non-specific resistance against infections. This
kind of resistance is exhibited with following two important functions

• Primary function

This function of complement proteins is considered primary as these activated proteins

used to kill or lyse infectious agents like bacteria. For this complement proteins get
deposited on the surface of infectious agents which make hole in those cells for their lysis
or killing.

• Secondary or Effector functions

These functions of complement proteins effect on other functions of immune system.

These functions of complement proteins are as follows

1) Opsonization
It is the process by which phagocytosis of microbes by immune cells get enhanced. Those
proteins which undertake this process are called as opsonins. For example, some

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BT-302 Immunology: Complement System

complement proteins like C3b, iC3b and C4b are good opsonins. These proteins attach to
microorganism first making a complex. This complex of complement protein & microbe
bind with complement receptor on the surface of phagocytic cell as a result phagocytosis
of microbe occur by phagocytes (Fig.11.1)

Figure 11.1: The opsonization function of complement proteins

2) Chemotaxsis

The phenomenon by which phagocytic cells move towards the site of infection is called as
chemotaxsis, and the molecules involved in this process are called as chemotactic factors. The
complement proteins are considered as good chemotactic factors like C5a. These act as potent
activator of neutrophils, basophils & macrophages. These cause the induction of adhesion
molecules on surface of blood vessel’s endothelial cells, which bind the phagocytic cells on it
(Fig.11.2). One of the important complex of complement proteins called as membrane attack
complex (MAC) composed of C5b, C6 and C7 also act as good chemotactic factor.

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Figure 11.2: The chemotactic function of complement proteins

3) Anaphylaxsis

Anaphylaxsis is the form of immune response in which exaggerated response is generated

against allergen. The molecules which produced such response are called as anaphylatoxins. The
complement proteins like C4a, C3a & C5a act as potent anaphylatoxins. These molecules cause
basophils and mast cells degranulation as a result various mediators or cytokines are generated
(Fig 11.3). Those mediators involved in smooth muscles contraction, vasodilatation and
bronchoconstriction.

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Figure 11.3: The anphylatoxsis function of complement proteins

Complement Activation

Complement proteins are activated through a series of cascading reactions which are called as
complement activation. Followings are the important pathways for complement activation

1) Classical Pathway
2) Alternate Pathway
3) Mannose Bindning Lectin (MBL) Pathway

These pathways generate the activated form of complement like C5 which leads towards a
pathway called as common pathway as a result the membrane attack complex is generated for the
lysis of microorganism (Fig 11.4)

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Figure 11.4: The pathways of complement activation

Classical Pathway

This pathway of complement activation is also called as antibody dependent complement

activation. As in this pathway, antibody bind to microbe which links as the first molecule of
classical pathway. This pathway begins with C1 activation which is a multi subunit protein
containing three sub proteins

i) C1q,
ii) C1r
iii) C1s

C1 protein binds with antibody on the surface of microorganism

Followings are important events of this pathway

1) Activation of C1

In this event, there is binding of C1q to Fc portion of Immunoglobulin (IgM& IgG) which have
bounded with antigens on bacterial surface. Binding of C1q in turn activates another subunit C1r

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and ultimately C1s (Fig 11.5). The activated C1qrs act as enzyme for C4 to cleave it in C4a &
C4b

Figure 11.5: Initiation of classical pathway with C3 convertase generation

2) Generation of C3 convertase

As C1qrs is activated, this complex also act on C2. As a result, C2 cleaves into two subunits C2a
& C2b. C2a binds on bacterial surface with C4b which is also a subunit of C4 generated after its
cleavage. This complex of C4b & C2a on bacterial surface is called as C3 convertase which
further acts on C3 to c3a and c3b (as shown in Fig. 11.5).

3) Generation of C5 convertase

In this stage, the activated C3 convertase (C4b&C2a) act on C3 to convert into C3a & C3b. C3a
moves into microenvironment for other immunological functions while C3b binds with C4b
&C2a. As a result, a complex of C4bC2aC3b is formed. This complex is called as C5 convertase
for the conversion of C5 into C5a & C5b.

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Figure 11.6: An overview of events during classical pathway of complement activation

Alternative Pathway of Complement Activation

This pathway of complement activation is antibody independent complement activation as there

is no need of antibodies for this. This is initiated by direct conversion of C3 into C3a & C3b.
Moreover, there is need of various serum proteins & factors like Factor B, D & Mg++ ions.
Generally, in serum there is low level spontaneous hydrolysis of C3 to produce C3i. In serum,
then factor B binds with C3i to form a complex of C3iB. This complex become susceptible to
another serum factor called as factor D which cleaves B into Bb. As a result, a complex C3iBb is
formed which acts as C3 convertase for conversion of C3 into C3a & C3b. The resulting C3b
reacts again with factor B and become susceptible to factor D, There is continuous formation of
C3bBb which acts as C3 convertase for cleavage of C3 (as shown in fig.11.7).

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Figure 11.7: Conversion of C3 into C3b during alternative pathway

As a result of this C3b amplification loop, there is more production of C3b. This amplification
loop of C3b is triggered by various molecules of microorganisms like lipopolysaccgrides (LPS)
of Gram negative bacteria, cell wall of bacteria & yeasts. These are also called as activator of
alternative pathway. Physiologically, the autonomus activation of C3 is controlled by following
mechanisms

1) By Decay accelerating factor (DAF)

2) Control of C3b amplification is done by a molecule called as Decay accelerating
factor (DAF), as a result the formation of C3 convertase is blocked (Figure 11.8)

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Figure 11.8: Control of spontaneous activation of C3 via DAF

2) By dissociating C3 convertase after cleavage of Bb from C3b

In this control mechanism, the spontaneous amplification of C3b loop is controlled by

enzymatic degradation of C3b by serum factor i.e factor H & I (Figure 11.9)

Figure 11.9: Dissociation of C3 convertase after cleavage of Bb from C3b

Clinically, the deficiency of Factor H & I in serum leads to increased susceptibility of patients to
various infections due to non-availability of active C3.

In alternative pathway, the activated C3b is stabilized on the surface of infectious agent which is
also called as activation surface by a protein called as protector (P) protein. This protector
protein is absent on homologous cells of body (Fig. 11.10)

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Figure 11.10: Stabilization of C3 on Activator surface by Protector Protein

As C3b is stabilized on activator surface, two molecules of C3b, Bb and protector protein (P)
form a complex called as C5 convertase which is the fate of alternative pathway (Fig.11.11).

Figure 11.11: Structure of C5 convertase of alternative pathway

Lectin Pathway of Complement Activation

This pathway is also called as antibody independent complement activation like alternative
pathway. However, it is initiated by mannose binding Lectins (MBL) on bacterial surface with
mannsoe containing polysaccharides (Mannans). After binding of MBL on bacterial surface,
there is association of two serine proteases called mannose-associated serine proteases (MASP).
There are following two types of MASPs

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1. MASP-1
2. MASP-2

In this pathway, MASP-1 corresponds like C1r & MASP-2 like C1s of classical pathway
respectively. The MBL acts like C1q of classical pathway. The activation of MASPs happen
after the formation of MBL/MASP-1 & MASP-2 tri molecular complex. This complex cleaves
C4 & C2 into C4b & C2a respectively for the formation of C3 convertase (Fig.11.12)

Figure 11.12: Mannose binding Lectin (MBL) pathway for complement activation

This C3 convertase cleaves of C3 into C3a &C3b. The C3b binds with C4b & C2a for the
generation of C5 convertase (C4b, C2a & C3b complex). While C3a move to microenvironment
for other immunological functions. The biological activities of C4a, C2b & C3a & regulatory
proteins of MBL pathway are same like classical pathway.

Lytic (Common) Pathway

This pathway is also called as Membrane attack complex (MAC) pathway as it is involved in the
lysis of infectious agents. C5 convertase is generated from all three following pathways

• Classical:C4b2a3b

• Alternative:C3bBb3b

• Lectin:C4b2a3b

This C5 convertase converts C5 into C5b &C5a. This C5b rapidly associates with other
complement proteins like C6 &C7 and insert into membrane. Subsequently, another protein

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called C8 binds with these membrane inserted proteins followed by several molecules of C9.
These C9 proteins make pore in the membrane which leads towards leakage of cellular contents
for cytolysis. The C5bC6C7C8C9 is called as membrane attack complex (MAC) (Fig 11.13)

Figure 11.13: The lytic (common) pathway of complement activation

Biological Active Products of Complement

The complement system provides specific & non-specific resistance against infections. As as
result of complement activation, there is production of various biological active molecules which
are involved in resistance, anaphylaxis & inflammation. Followings are the important biological
active products of complement

Kinnin production

C2b which is produced in classical pathway is called as Pro-Kinnin. This Pro-Kinnin is activated
by serum factor called as Plasmin in to Kinnin which is a potent biological active compound.
The excess C2b production cause undesirable effects like smooth muscles contraction and
vasodilatation

Anaphylatoxins

Anaphylaxsis is the form of immune response in which exaggerated response is generated

against allergen. The molecules which produced such response are called as anaphylatoxins. The
complement proteins like C4a, C3a & C5a act as potent anaphylatoxins. These molecules cause
basophils and mast cells degranulation as a result various mediators or cytokines are generated

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BT-302 Immunology: Complement System

(Fig 11.3). Those mediators involved in smooth muscles contraction, vasodilatation and
bronchoconstriction.

Chemotactic factors

The phenomenon by which phagocytic cells move towards the site of infection is called as
chemptaxsis, and the molecules involved in this process are called as chemotactic factors. The
complement proteins are considered as good chemotactic factors like C5a. These act as potent
activator of neutrophils, basophils & macrophages. These cause the induction of adhesion
molecules on surface of blood vessel’s endothelial cells, which bind the phagocytic cells on it
(Fig.11.2). One of the important complex of complement proteins called as membrane attack
complex (MAC) composed of C5b, C6 and C7 also act as good chemotactic factor.

Opsonins

It is the process by which phagocytosis of microbes by immune cells get enhanced. Those
proteins which undertake this process are called as opsonins. For example, some complement
proteins like C3b, iC3b and C4b are good opsonins. These proteins attach to microorganism first
making a complex. This complex of complement protein & microbe bind with complement
receptor on the surface of phagocytic cell as a result phagocytosis of microbe occur by
phagocytes (Fig.11.1)

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