The Complement System

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Introduction

• Major component of the innate immune system

• Consists of about 30 proteins in the blood stream that activate one
another in response to pathogenic attack.
• Discovered in the 1890s by Jules Bordet as a heat-labile substance in
normal plasma whose activity could ‘complement’ the bactericidal
activity of immune sera.
• The proteases of the complement system are synthesized (most of
them in the liver) as inactive pro-enzymes, or zymogens.
• Become enzymatically active only after proteolytic cleavage, usually by
another complement protein.
• The activation occurs in an enzyme cascade fashion and generally
involve activation, amplification and a terminal effect.
• There are three known pathways in which this cascade takes place.
Classical, alternative and lectin.
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S P O N S E
 Activation
Main differentiating factor among the pathways

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Classical pathway

Initiated when antibodies bind to the surface antigens on a

pathogen.

Antibodies involved are Ig G or Ig M.

It takes at least two antigen-bound antibodies in close

proximity to each other, to attract the first complement
protein, activating the pathway.
This applies mostly to Ig G since they circulate in monomeric
structure as compared to Ig M which is pentamer.
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S P O N S E
 Classical pathway
• C1 — the first protein. Comprises of large C1q molecule and two molecules each of C1s and C1r.

• C1q — is the globular head portion that binds to the Fc portions of antigen-bound antibody.
Conformationational change in C1q leads to auto-catalytic conversion of C1r portion into active
serum protease.

• C1r — cleaves C1s into it's active form.

• C1s — cleaves C4 into: C4a and C4b

• C4b — attaches covalently to the antigen-antibody complex and has a binding site for C2a.

• C2a — binds to C4b after being cleaved off C2 by the same C1s.

• C4bC2a — bimolecular complex serving as a C3 convertase via the C2a portion.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S P O N S E
 Alternative pathway
Activated by components on the surface of pathogens. Include LPS.
C3(H2O) — always present in serum at low levels due to the spontaneous hydrolysis of C3. This means
that the alternative pathway is constantly active at very low levels.

Factor B — binds to C3(H2O) and becomes susceptible for cleavage by factor D.

Factor D — cleaves Factor B into Bb and Ba. Bb remains attached to C3(H2O) and is the active portion of
the biomolecule C3(H2O)Bb. The bimolecular complex is referred to as a C3 initiation complex.

C3(H20)Bb — Cleaves C3 into C3a and C3b.

C3b — Binds to host surfaces and only persists in the presence of an appropriate activator (LPS).

C3bBb — is the predominant C3 convertase similar to C4bC2a in classical pathway. It is formed by binding
of Factor B to pathogen-bound C3b. As expected, Factor D cleaves off Ba from Factor B, and the Bb
remains to form the biomolecule complex.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Properidin — stabilizes
the C3bBb

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Lectin pathway

Activated by PRRs namely, MBL (mannose-binding lectin) and ficolins

binding PAMPs.

The PAMPs involved are mannose sugars and other oligosaccharides

on the pathogenic surfaces.

MBL — similar to C1q in classical pathway. Complexed with MBL-

associated serine proteases (MASPs)-1 and -2 which are
functionally and structurally similar to C1r and C1s respectively.

MASPs — are activated when MBL binds to pathogenic surface and in

turn cleave of C2 and C4 ultimately generating C4bC2a.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S P O N S E
 Amplification
Mainly consists of conversion of C3 and C5 by their
respective convertases

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Classical
and lectin
C4bC2a — cleaves C3 into
C3a and C3b.

C3b — associates with

C4bC2a forming the C5
convertase, C4bC2aC3b.

C4bC2aC3b — cleaves C5
into C5a and C5b

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Alternate
pathway
C3bBb — cleaves C3 into
C3a and C3b.

C3b — associates with

C3bBb to form C5 convertase,
C3bBbC3b. It may also
covalently bind to surface to
attract Factor B and D for
additional cleavage of C3.
Provides an amplification loop.

C3bBbCb — cleaves C5 into

C5a and C5b

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Termination
Complement activation, regardless of the pathway,
converges on the generation of three broad effector
pathways that serve to enable the complement to fulfill
its physiological imperatives in host defense

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Recruitment of
other components
of the immune
system
Initiated by anaphylotoxins, C3a,
C4a and C5a which are highly
related proinflammatory
molecules.

C5a is the most potent followed by

C3a while C4a has been shown to
be considerably weak.

C5a — chemotactic factor for

neutrophils and also stimulates
activity of macrophages.

C3a — causes degranulation of

mast cells and basophils.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Membrane
attack
complex
generation of porous complexes on
pathogenic surfaces that result in lysis of
bacteria.

C5b — forms the foundation by

associating with C6 C7 C8 sequentially.

C8 — first to penetrate the host

membrane before multiple C9 molecules
are recruited to the site.

C9 — molecules polymerize forming a

pore in the membrane with a maximum
diameter of 10 nm.

MAC — C5b-9 complex that rapidly

increases intake of water and Ca2+
accompanied with leakage of pathogen
cellular content.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Opsonization of
pathogens
C3b — coats the surfaces of
pathogens and enhance
phagocytosis by
macrophages.

CR1 — is a receptor found on

macrophages that adheres to
pathogen bound C3b.

Involved in clearance of
immune complexes

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S P O N S E
 Action of Complement on
Adaptive Immunity
• Besides acting in innate immunity, complement also influences
adaptive immunity i.e.
 Opsonization of pathogens by complement facilitates their uptake by
phagocytic antigen-presenting cells enhancing the presentation of
pathogen antigens to T cells.
 B cells express receptors for complement proteins that enhance their
responses to complement-coated antigens
• In addition, several of the complement fragments can act to influence
cytokine production by antigen-presenting cells, thereby influencing
the direction and extent of the subsequent adaptive immune
response.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
 Regulation of complement
activation
• Regulation confines activity to appropriate pathogenic surfaces, and prevent collateral damage to
healthy host tissues following generation of potent effectors.
• C1-INH (C1 inhibitor ) — binds to activated C1r and C1s, limit classical pathway activation.
• Deficiency of C1-INH is basis of hereditary angioedema

• Factor I — a serine protease that constitutively cleaves C3b and C4b into inactive fragments,
such as iC3b, C3c, C3dg.

• Factor H and CR1 — are membrane proteins on host cells that serve as co-factors for the
proteolytic activity of Factor I. These prevents the non-specific degradation of C3b, such as in the
case of proper complement activation.

• DAF(decay-accelerating factor) — comes into play when C3b deposition cannot be controlled.
• Inhibits assembly of new convertase and shorten the half-life of preformed convertase.
• Blocks association of Factor B with C3b.
• Accelerates dissociation of Bb from C3b.

• CD59 — Inhibits polymerization of MAC thereby preventing MAC assembly

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
Complement evasion/subversion strategies of
microorganisms

Complement is an evolutionarily ancient component of host defense.

And, given that it has coevolved with pathogens for millions of years, its time enough for pathogens to
develop mechanisms to evade complement activity.

Gram-positive bacteria — Inhibit MAC assembly simply by virtue of having a thick cell wall.

Staphylococcus aureus produces a membrane protein, Staphylococcal protein A (SpA) — binds to the
Fc region of IgG,
limiting complement activation via the classical pathway by interfering with the binding of C1q
also inhibiting Fc-receptor-mediated phagocytosis

Borrelia burgdorferi — encodes a 80 kDa surface protein that shares functional similarities with human
CD59

Neisseria family of pathogens — use protein mimicry to recruit Factor H to its surface and evade
persecution.

Laboratory Training for FieldEEpidemiologists

P I D E M I C A L E R T A N D R E S P O N S E
Laboratory Training for FieldEEpidemiologists
P I D E M I C A L E R T A N D R E S J
P ALLERGY
O N S E CLIN IMMUNOL . APRIL 2004