COMPLEMENT

SYSTEM

[Link] J
 HISTORY
Jules Bordet at the Institut Pasteur in Paris (1890s) –
“Alexins”-Greek’ ‘to ward off’
⚫ Paul Ehrlich coined “complement “,

defining it as “the activity of blood serum

that completes the action of antibody.”
 INTRODUCTION
It is named “complement system” because it was first identified as
a heat-labile component of serum that “complemented” antibodies
in the killing of bacteria.

- “group of proteins normally found in serum in inactive form,but when activated they augment

the immune responses”.

 mainly synthesized by hepatocytes

 Also produced by blood monocytes, tissue macrophages and epithelial cells of the
gastrointestinal and genitourinary tract.

 constitute 5% (by weight) of the serum globulin fraction

-denatured by heating at 560C – inactivated serum

 The Complement system is the major effector of humoral branch of immune system.
 5)Components are designated by
numbers (E.g. ; C1 – C9) or letters
(E.g. : Factor D).

Those complexes that have

enzymatic activity are designated by a
bar over the number or symbol (e.g.,
C4b2a, C3bBb).
 THE COMPONENTS OF COMPLEMENT SYSTEM

Over 30 serum and cell surface proteins:

- Complement components
(in serum inactive, activated sequentially as a cascade)
- Complement receptors
(cell surface, recognize activated components)
- Regulatory proteins and properdin system
(both in serum and cell surface, inhibit activated
components)
 Complement proteins:Made as zymogens/proenzymes
(inactive) - activation by cleavage(exposes the active site).
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(Fc of Ab), triggering
specific cell functions, inflammation, and secretion of immunoregulatory molecules
■ Immune clearance, which removes immune complexes from the circulation and deposits them
in the spleen and liver
 SOME DEFINITIONS

• C-activation: alteration of C proteins such that they interact with the next
component
• C-fixation: utilization of C by Ag-Ab complexes
• C-inactivation: denaturation (usually by heat) of an early C-component
resulting in loss of hemolytic activity
• Convertase/esterase: altered C-protein which acts as a proteolytic enzyme
for another C-component
THREE PATHWAYS FOR COMPLEMENT ACTIVATION:

 1. Classical Pathway-antibody dependent pathway and triggered by

formation of soluble antigen-antibody complex or by binding of the antibody
to the antigen present on the target cell surface.
 2. Alternative Pathway- antibody independent pathway stimulated by
antigen directly eg. Bacterial cell surface components
 3. Lectin or MBL Pathway-Also antibody independent but resembles
classical pathway.

Of these pathways, the lectin and the alternative pathways are more
important (antibody not present during first attack) - therefore, participants in
the innate arm of the immune system
 Pathways for activation require multiple steps categorized as
 STAGES OF COMPLEMENT ACTIVATION

⚫ Four main stages in the activation of complement by any pathway are

 Initiation of the pathway

 Formation of C3 convertase
 Formation of C5 convertase
 Formation of membrane attack complec(MAC)
THE CLASSICAL PATHWAY
C1-C9

 Part of adaptive immune system

 Relies on Ag – Ab complex to get activated
 The classical pathway is initiated by:

[Link] binding to the pathogen

2.C1 proteins binds to the Fc of Ab.

IgM(pentameric)>IgG3>IgG1>IgG2(monomeric).
Planar IgM Staple IgM

C1 Protein

C1q- 18 polypeptides(hexamer)
6 arms with globular heads-
Binds Fc on IgG or IgM to get
activated

2 C1r + 2 C1s are activated by activated

C1q in the presence of Ca2+.
 C14b

Mg2+

C14b2a

(C1s esterase)
C14b2a3b
Cleavage of C3 reveals a thioester bond and
it will bind to cell surface of pathogen.
 MBL is an acute phase protein produced in
inflammatory responses

similar to C1q in structure and

function
 Salmonella, Listeria, and Neisseria strains, as well
as Cryptococcus neoformans and Candida albicans.

(MBL/MASP-C4b2a)
 C3,C5-9 +
(Properdin pathway)Factor B,D,P-
Cell surface
constituents foreign to properdin
host-
bacterial endotoxin

(unstable thioester bond)

.
⚫ C3b has two important functions:

(1)It combines with other

complement components to
generate C5 convertase, the
enzyme that leads to the production
of the membrane attack complex
and

(2)it opsonizes bacteria because

phagocytes have receptors for
C3b on their surface.

C3b is an opsonin that bind both to

bacteria and phagocytes

Opsonization increases
phagocytosis by 1,000 fold
MEMBRANE ATTACK
COMPLEX
 Cleavage of C5 into C5a and C5b.
 C5 (structurally homologous to C3 and C4, lacks internal thioester bond )
 C5b initiates formation of MAC (complex of C5b, C6, C7, C8 and multiple C9
molecules ) binds to C6, and C7 , recruits C8 and complex penetrates more
deeply into the membrane.
 C9, a pore-forming molecule with homology to perforin. The complex of C5b678
forms a nidus for C9 binding and polymerization
 Penetrates membrane bilayers to form pores
 Disrupt the osmotic barrier, leading to swelling and lysis of susceptible cells

Abbas [Link]&Molecular immunology 6th edition

 10 nm
MAC FORMATION
 C4b2a
C4b2a3b
 EFFECTOR FUNCTIONS OF
COMPLEMENT SYSTEM

1. Facilitates Opsonization :C3b & C4b; enhance phagocytosis

Extremely important when pathogen carries a

capsule.
[Link] LYSIS
3. IMMUNE COMPLEX
CLEARANCE-
4. Inflammatory response and chemotaxis-
C5,6,7 complex  attract neutrophils
C5a – enhance adhesiveness of neutrophils to the
endothelium
and C3b
[Link] (C3a, C4a, C5a)
Cause degranulation o f m a s t c e l l s
b i n d d i r e c t l y t o s m o o t h muscles o f bronchioles  bronchospasm

[Link] NEUTRALIZATION
-coat on viral surface –block their attachment sites
-C3 mediated opsonization
-lysis of enveloped virus by activation of classical pathway in most and by alternative or lectin pathway in others –EBV,Rubella virus,etc.
OVERVIEW
REGULATION OF COMPLEMENT
SYSTEM
1. C1 inhibitor
• Important regulator of classic pathway
• A serine protease inhibitor (serpin)
• Irreversibly binds to and inactivates C1r and
C1s, as well as MASP in lectin pathway

2. Factor H
• Regulate alternative pathway
• Competes Factor B
• Binds to C3b on host cells to prevent activation
on them
• Reduce amount of C5 convertase
available(displace C3b from C5 convertase)
3. Properdin
• Protects C3b and stabilizes C3 convertase

4. Factor I
• Cleaves C3b and C4b inactivates C3b and C4b

5. Decay accelerating factor (DAF)

• Glycoprotein on surface of human cells
• Prevents assembly of C3bBb (competes with Factor B) or
accelerates disassembly of preformed convertase  no
formation of MAC
• Acts on both classical and alternative
6. C4b-binding protein (C4BP)
• Inhibits the action of C4b in classical pathway
• is a cofactor for factor I

7. Complement Receptor 1 (CR-1)

• Co-factor for factor I

8. Protectin (CD59-membrane bound protein) and

Vitronectin (plasma protein)

• Inhibits formation of MAC by binding C5b678

• Present on “self” cells to prevent complement
from damaging them
MEASURING THE 50% HAEMOLYTIC
COMPLEMENT (CH 50 ) ACTIVITY OF SERUM
Individual complement components can be quantified
The CH50 is a screening assay ,it tests the functional capability of serum complement
components of the classical pathway to lyse sheep red blood cells (SRBC) pre-coated
with rabbit anti-sheep red blood cell antibody (haemolysin).
When antibody-coated SRBC are incubated with test serum, the classical pathway of
complement is activated and haemolysis results. If complement component are absent,
the CH50 level will be zero; if one or more components of the classical pathway are
decreased, the CH50 will be decreased.
A fixed volume of optimally sensitised SRBC is added to each serum dilution. After
incubation, the mixture is centrifuged and the degree of haemolysis is quantified by
measuring the absorbance of the haemoglobin released into the supernatant at 540nm.
The amount of complement activity is determined by examining the capacity of
various dilutions of test serum to lyse antibody coated SRBC. s.
 CLINICAL ASPECTS OF COMPLEMENT
1. Deficiency of C5-C8 & Mannan-binding lectin
• Predispose to severe Neisseria bacteremia
2. Deficiency of C3
• Severe, recurrent pyogenic sinus & resp. tract infections
3. Deficiency of C1 esterase inhibitor
• Angioedema inc. capillary permeability and
edema
4. Deficiency of DAF
• Increased complement-mediated hemolysis 
paroxysmal nocturnal hemoglobinuria
5. Transfusion mismatches
• Activation of complement  generate large amounts of
anaphylatoxins & MAC
 red cell hemolysis
[Link] liver disease
• Deficient complement proteins 
predispose to infection with pyogenic bacteria
7. Factor I deficiency

• Low levels of C3 in plasma due to unregulated activation of alternative

pathway recurrent bacterial infections in children
• Mutations in factor I gene implicated in
development of Hemolytic Uremic Syndrome
 CHOUAKI BENMANSOUR, N., CARVELLI, J.
AND VIVIER, E. (2021), COMPLEMENT
CASCADE IN SEVERE FORMS OF COVID-19:
RECENT ADVANCES IN THERAPY. EUR. J.
IMMUNOL., 51: 1652-1659.
HTTPS://[Link]/10.1002/EJI.202048959
The complement system is an essential component of the innate
immune system. The three complement pathways (classical, lectin,
alternative) are directly or indirectly activated by the SARS-CoV-2
(severe acute respiratory syndrome coronavirus 2). In the most
severe forms of COVID-19, overactivation of the complement
system may contribute to the cytokine storm, endothelial
inflammation (endotheliitis) and thrombosis. No antiviral
drug has yet been shown to be effective in COVID-19.
Therefore, immunotherapies represent a promising
therapeutic in the immunopathological phase (following the
viral phase)of the disease. Complement blockade, mostly
C5a-C5aR axis blockade, may prevent acute respiratory
distress syndrome (ARDS) from worsening or progression to
death. Clinical trials are underway.
Anti –complement drugs
Figure. Coronavirus and complement. A, Eculizumab inhibits C5, preventing breakdown into C5a and C5b, which
is an integral component of the membrane attack complex (MAC). B, In humans, atypical hemolytic-uremic
syndrome (aHUS), early treatment with eculizumab reverses organ dysfunction.
THANK
U