What is complement system ?

Complement refers to a collection of over 30 heat-labile proteins found in human blood

plasma. The proteins act in a cascading fashion to lyse cell membranes, augment
phagocytosis, and produce inflammatory peptides. These activities were said to "complement“
the other antibacterial activities of the host; hence the name complement.

 Complement is a multiprotein network of plasma proteins and cell surface receptors.

 Complement is an evolutionary conserved system.
 Complement participates in innate and adaptive immunity.
 Complement dysregulation and deficiencies are connected to disease.
 Objectives of Complement System

(1) Defends against bacterial infections by facilitating and enhancing phagocytosis,

chemotaxis, activation of leukocytes, and lysis of bacteria;

(2) Bridges innate and specific immune functions by enhancing antibody responses and
immunologic memory;

(3) Disposes of wastes such as dead host cells and immune complexes, the products of
inflammatory injury.
Sequential activation of complement components occurs via one
of three pathways:

1. Classic pathway
2. Lectin pathway
3. Alternative or Properdin pathway

• Lectin and alternative pathways are more important the first time
we are infected by microorganisms because antibody required
to activate the classic pathway is not yet present
 Initiation complex for complement activation
1. Classic pathway
• Part of acquired or adaptive immunity
• Activated by Ag-Ab complexes
• Immunoglobulins involved: IgM and IgG
(except IgG4)
• Involves activation of C1
 Composed of C1q, C1r, and C1s  binds
to Fc portion of IgG and IgM
 Requires calcium for activation
2. Lectin pathway
• Also known as the MBL Pathway
• Activated by binding of mannose-binding lectin
(or mannose-binding protein) to surface of
microbes bearing mannan (polymer of the sugar
mannose) in a calcium dependent manner
• Binding causes activation of MASP
(MBPassociated serine proteases)  cleave C2
and C4
The lectin pathway is triggered by a plasma protein called mannose-binding lectin (MBL), which recognizes terminal
mannose residues on microbial glycoproteins and glycolipids, similar to the mannose receptor on phagocyte membranes
described earlier (see Figure 1A). MBL is a member of the collectin family (discussed later) with a hexameric structure
similar to the C1q component of the complement system. After MBL binds to microbes, two zymogens called MASP1
(mannose-associated serine protease 1, or mannan-binding lectin-associated serine protease) and MASP2, with similar
functions to C1r and C1s, associate with MBL and initiate downstream proteolytic steps identical to the classical pathway.
The classical pathway, so called because it was discovered first, uses a plasma protein called C1q to detect antibodies
bound to the surface of a microbe or other structure . Once C1q binds to the Fc portion of the antibodies, two
associated serine proteases, called C1r and C1s, become active and initiate a proteolytic cascade involving other
complement proteins. The classical pathway is one of the major effector mechanisms of the humoral arm of adaptive
immune responses. Innate immune system soluble proteins called pentraxins, can also bind C1q and initiate the
classical pathway.
The alternative pathway, which 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. Recognition of microbes by any of the three complement pathways results in sequential recruitment and
assembly of additional complement proteins into protease complexes. One of these complexes, called C3 convertase, cleaves the
central protein of the complement system, C3, producing C3a and C3b. The larger C3b fragment becomes covalently attached to
the microbial surface where the complement pathway was activated. C3b serves as an opsonin to promote phagocytosis of the
microbes. The smaller fragment, C3a, is released and stimulates inflammation by acting as a chemoattractant for neutrophils.
Complement Activation
b
C3b binds other complement proteins to form a protease called C5 convertase that cleaves C5, generating a released peptide (C5a) and a larger fragment
(C5b) that remains attached to the microbial cell membranes. C5a is also a chemoattractant; in addition, it induces changes in blood vessels that make them
leak plasma proteins and fluid into sites of infections. C5b initiates the formation of a complex of the complement proteins C6, C7, C8, and C9, which are
assembled into a membrane pore called the membrane attack complex (MAC) that causes lysis of the cells where complement is activated.

The complement system is an essential component of innate immunity, and patients with deficiencies in C3 are highly susceptible to recurrent, often lethal,
bacterial infections. Genetic deficiencies in MAC formation (the terminal product of the classical pathway) increase susceptibility to only a limited number
of microbes, notably Neisseria bacteria, which have thin cell walls that make them especially susceptible to the lytic action of the MAC.
Membrane Attack Complex formation
Functions of Complements
Key Complement Proteins and Their Functions
 Biologic Effects

1. Opsonization 4. Cytolysis (MAC)

• C3b & C1q; enhance phagocytosis • Disrupt the membrane & the entry of
2. Chemotaxis water and electrolytes into the cell
• C5a and C5,6,7 complex  attract 5. Enhancement of antibody production
neutrophils • Binding of C3b to its receptors on the
• C5a – enhance adhesiveness of surface of activated B cells enhanced
neutrophils antibody production
to the endothelium
3. Anaphylatoxin (C3a, C4a, C5a)
• Cause degranulation of mast cells
• Bind directly to smooth muscles of
bronchioles  bronchospasm
1. C1 inhibitor (C1-INH)
• 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
• Reduce amount of C5 convertase available
• With both cofactor activity for the factor I-
mediated C3b cleavage, and decay accelerating
activity against C3bBb (C3 convertase)
3. Properdin
• Protects C3b and stabilizes C3 convertase.
4. Factor I
• Cleaves cell-bound or fluid phase C3b and
C4b  inactivates C3b and C4b
5. Decay accelerating factor (DAF)
• Glycoprotein on surface of human cells
• Prevents assembly of C3bBb 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
• Splits C4 convertase and is a cofactor for
factor I
7. Complement Receptor 1 (CR-1)
• Co-factor for factor I, together with CD46
8. Protectin (CD59) and Vitronectin (S protein)
• Inhibits formation of MAC by binding
C5b678
• Present on “self” cells to prevent
complement from damaging them
Regulation of complement system
 Clinical Aspects
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
6. Autoimmune diseases
• Immune complexes bind complement low
complement levels + activate
inflammation  tissue damage
7. Severe liver disease
• Deficient complement proteins 
predispose to infection with pyogenic
Bacteria
8. 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
Regulation of Complement System

1. C1 inhibitor (C1-INH)
• 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
• Reduce amount of C5 convertase available
• With both cofactor activity for the factor I-mediated C3b cleavage, and
decay accelerating activity against C3bBb (C3 convertase)
3. Properdin
• Protects C3b and stabilizes C3 convertase.
4. Factor I
• Cleaves cell-bound or fluid phase C3b and C4b  inactivates C3b and
C4b.
5. Decay accelerating factor (DAF)
• Glycoprotein on surface of human cells
• Prevents assembly of C3bBb 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
• Splits C4 convertase and is a cofactor for factor I
7. Complement Receptor 1 (CR-1)
• Co-factor for factor I, together with CD46
8. Protectin (CD59) and Vitronectin (S protein)
• Inhibits formation of MAC by binding C5b678
• Present on “self” cells to prevent complement from damaging them
Complement Fixation Test