Complement system

Deo Okoche
• The complement system is made up of a large
number of distinct plasma proteins that react with
one another to;
 Opsonize pathogens
 Lyse (kill) pathogens
 Induce a series of inflammatory responses that help to fight
infection
• A number of complement proteins are proteases
called zymogens activated by proteolytic cleavage
• These Zymogens include; C1, C4, C2, C3, C5, C6,
C7, C8, and C9
• The products of cleavage reactions are designated
by added lower-case letters, the larger fragment
being designated b and the smaller a (i.e C4a &
C4b)
• The precursor zymogens are widely distributed
throughout body fluids and tissues without adverse
effect
• At sites of infection, they are activated locally and
trigger a series of potent inflammatory events
• The complement system activates through a
triggered-enzyme cascade
• In such a cascade, an active complement
enzyme generated by cleavage of its zymogen
precursor then cleaves its substrate, another
complement zymogen, to its active enzymatic
form
• This in turn cleaves and activates the next
zymogen in the complement pathway
• In this way, the activation of a small number of
complement proteins at the start of the pathway
is hugely amplified by each successive
enzymatic reaction,
 Resulting in the rapid generation of a large complement
response
 Pathways of the complement system
• 3 distinct pathways through which complement can be
activated on pathogen surfaces
• All depend on different molecules for their initiation, but
they converge to generate the same set of effector
molecules

Source: Immunology by JaneWAY

• The classical pathway can be initiated by;
 Binding of C1q, the first protein in the complement cascade,
directly to the pathogen surface
 Binding of C1q to antibody: antigen complexes, and is thus a
key link between the effector mechanisms of innate and
adaptive immunity
• The mannan-binding lectin pathway (MB-lectin
pathway)
 Initiated by binding of the mannan-binding lectin, a serum
protein, to mannose-containing carbohydrates on bacteria or
viruses
• The alternative pathway can be initiated when a
spontaneously activated complement component
binds to the surface of a pathogen

Each pathway follows a sequence of reactions to

generate a protease called a C3 convertase
Early steps of complement
activation
 The classical pathway
• This pathway is initiated by C1q which is part of
complement protein complex C1
• A single C1q molecule bound to two molecules
which include; C1r and C1s
Molecule of C1q

Source: Immunology by JaneWAY

• Binding of more than one of C1q heads to a Fc of
Antibody causes a conformational change in the
(C1r:C1s)2 complex
• Leads to activation of an autocatalytic enzymatic
activity in C1r;
• The active form of C1r then cleaves its associated
C1s to generate an active serine protease
• Once activated, the C1s enzyme acts on the next two
complement proteins (C4 and C2)
• Active C1s cleaves C4 and then C2 to generate two
large fragments, C4b and C2b which together form
C3 convertase
• C3 convertase cleaves C3 molecules to produce large
fragment of C3b and small one of C3a molecules
• C3b molecules that coat the pathogen
(Opsonization) surface while C3a initiates a local
inflammatory response
Formation of C3b in classical pathway

Source: Immunology by JaneWAY

 Summary
of classical
complement
pathway

Source: Cellular & Molecular

immunology by Abbas
Proteins of classical pathway complement activation & their functions

Source: Cellular &

Molecular immunology by
Abbas
 The Mannan-binding lectin (MBL) pathway
• This pathway uses a serum protein called mannan-
binding lectin (MBL) similar to C1q
• MBL binds specifically to mannose residues & some
sugars on many pathogen surfaces
• On vertebrate cells, however, mannose & other
sugars bound by MBL are covered by other sugar
groups, especially sialic acid
• Thus, MBL is able to initiate complement activation
by binding to pathogen surfaces only
• MBL, like C1q, is a six-headed molecule that forms
a complex with two protease zymogens, MASP-1 &
MASP-2
• Note: MASP-Mannan-binding lectin activating
serine protease
• When MBL complex binds to a Structure of MBL
pathogen surface, MASP-1 and
MASP-2 are activated to cleave C4
and C2
• C4b bound to C2b forms a C3
convertase
• C3 convertase cleaves C3 molecules
to produce large fragment of C3b and
small one of C3a molecules Source: Immunology by JaneWAY

• C3b molecules that coat the pathogen

(Opsonization) surface while C3a
initiates a local inflammatory
response
Summary of
MBL
complement
pathway

Source: Cellular & Molecular

immunology by Abbas
 Alternative complement pathway
• It is called alternative pathway b’se it was
discovered as a second, or 'alternative,' pathway for
complement activation after the classical pathway
had been defined
• It can proceed on many microbial surfaces in the
absence of specific antibody
• In contrast to classical & MBL complement
activation, the alternative pathway does not depend
on a pathogen-binding protein for its initiation
Instead it is initiated through the spontaneous hydrolysis
of C3
It leads to the generation of a distinct C3 convertase
designated C3b,Bb
• Serum C3, contains an unstable thioester bond,
which subjects it to slow spontaneous hydrolysis to
yield C3a and C3b
• The C3b can bind to foreign surface antigens or
even to the host’s own cells
• The C3b present on the surface of cells can bind
another serum protein called factor B to form a
complex stabilized by Mg2+
• Binding to C3b exposes a site on factor B that
serves as the substrate for an enzymatically active
serum protein called factor D
• Factor D cleaves the C3b-bound factor B, releasing
a small (Ba) and a larger fragment (Bb)
• Ba diffuses away while Bb remains attached to
C3b & forms C3bBb complex which is C3
convertase of alternative pathway
• If C3b,Bb complex forms on the surface of host
cells, it is rapidly inactivated by complement-
regulatory proteins
• Pathogen surfaces don’t express complement-
regulatory proteins & favor binding of factor P
(properdin), which stabilizes the C3b,Bb
convertase activity
• C3 convertase cleaves more molecules of C3 to
C3a and C3b
C3a initiates a local inflammatory response
C3b molecules that coat the pathogen (Opsonize)
surface
Summary of
alternative
complement
pathway

Source: Cellular & Molecular immunology by Abbas

Proteins of alternative pathway complement activation & their functions

Source: Cellular & Molecular immunology by Abbas

 Late Steps of Complement Activation
• C3b4b2b complex in classical & MBL pathways and
C3bBbC3b complex in alternative pathway form C5
convertase
• C5 convertases generated by the alternative,
classical, or lectin pathway initiate activation of the
late components of the complement system
• This culminates in formation of the cytocidal
membrane attack complex (MAC)
• C5 convertases cleave C5 into a small C5a fragment
that is released and a large C5b fragment that
remains bound to the complement proteins deposited
on the cell surface
• C5a attracts immune cells to site of infection &
initiates inflammation
• C5b triggers the assembly of a complex of one
molecule each of C6, C7, and C8, in that order
• C7 and C8 undergo conformational changes that
expose hydrophobic domains that insert into the
pathogen membrane
• This complex causes moderate membrane damage
and also induces binding & polymerization of C9 by
exposure of a hydrophobic site
• Binding & polymerization of up to about 15 C9
molecules results into formation of membrane attack
complex (MAC)
• MAC generates channels which disrupt the pathogen
cell membrane, killing the bacterium by lysis
Late steps of complement activation and formation of the MAC

Source: Cellular & Molecular immunology by Abbas

Summary of the functions of the complement system
Proteins of the Late Steps of Complement activation & their functions

Source: Cellular & Molecular immunology by Abbas

Receptors on immune cells for Fragments of C3

Source: Cellular & Molecular immunology by Abbas

 Regulators of the
complement
system

Source: Cellular & Molecular

immunology by Abbas
 Detrimental effects of Complement activation
• Mutations in genes for complement regulators,
factor H and membrane cofactor protein have
been associated with atypical hemolytic uremic
syndrome (HUS)
 HUS is a disease characterized by hemolytic anemia (anemia
caused by destruction of red blood cells), acute kidney failure
(uremia), and a low platelet count (thrombocytopenia)
• A common single nucleotide polymorphism in
factor H (Y402H) has been associated with the
common eye disease age-related macular
degeneration
 Deterioration of the central portion of the retina
 Central portion is responsible for focusing central vision in
the eye, and it controls our ability to read, drive a car,
recognize faces or colors, and see objects in fine detail
• Mutations in the C1 inhibitor gene can cause
hereditary angioedema
 HAE is a rare, autosomal dominantly inherited blood disorder that
can result into rapid swelling of the face, hands, feet, genitals,
gastrointestinal tract and upper airways
• Complement activation in CNS can cause
Alzeimer’s disease (neurodegenerative disease
that causes progressive decline in memory,
thinking, language and learning capacity)
 Both glia and nerve cells in the brain can synthesize
complement components
 In Alzeimer’s disease patients there is formation of Neuritic
plaques
 Neuritic plaques are extracellular deposits of amyloid beta (Aβ)
in the grey matter of the brain associated with degenerative
neural structures and an abundance of microglia
 Aβ directly activates Alternative pathway & Classical pathway
by binding to C3 and the globular heads of C1q resulting into
formation of;
 C3a & C5a which are proinflammatory
 MAC which causes lysis of the neurones
• Complement activation is associated with
intravascular thrombosis & can lead to ischemic
injury to tissues
Antiendothelial antibodies against vascularized organ
transplants & immune complexes produced in
autoimmune diseases may bind to vascular endothelium
and activate complement,
Thereby leading to inflammation and generation of the
MAC with damage to the endothelial surface, which
favors coagulation
• Systemic vasculitis and immune complex
glomerulonephritis result from the deposition of
antigen antibody complexes in the walls of vessels
and kidney glomeruli
Thank you