COMPLEMENT
SYSTEM
• Complement system is a cascade of more
Introduction than 30 soluble and cell-bound proteins that
interact in a very specific way to enhance the
to hosts defense mechanism against foreign
cells. It plays a major role in cellular and
Complement humoral branch of immunity.
System • Complement is coined by Paul Ehrlich but
was extensively researched by Jules Bordet,
calling it alexine.
• While complement promotes opsonization
and lysis of foreign cells and immune
complexes, chronic activation can lead to
inflammation and tissue damage.
• It can be activated in three different ways,
involving the classical pathway, alternative
pathway and lectin pathway.
COMPLEMENT SYSTEM
• Most plasma complement proteins are synthesized in the liver, with exception of C1 components
which are produced in the intestinal epithelium, and factor D which are produced by adipose
tissues.
Classical Pathway Proteins
Serum protein Molecular weight (kD) Concentration (ug/mL) Function
C1q 410 150 Binds to Fc of IgM and IgG
C1r 85 50 Activates C1s
C1s 85 50 Cleaves C4 and C2
C4 205 300-600 Part of C3 convertase (C4b)
C2 102 25 Binds to C4b, forming C3 convertase
C3 190 1200 Key intermediate in all pathways
C5 190 80 Initiates membrane attack complex
C6 110 45 Binds to C5b in Mac
C7 100 90 Binds to C5bC6 in MAC
C8 150 55 Starts pore formation on membrane
C9 70 60 Polymerizes to cause cell lysis
COMPLEMENT SYSTEM
• Most plasma complement proteins are synthesized in the liver, with exception of C1 components
which are produced in the intestinal epithelium, and factor D which are produced by adipose
tissues.
Alternative Pathway Proteins
Serum protein Molecular weight (kD) Concentration (ug/mL) Function
Factor B 93 200 Binds to C3b to form C3 convertase
Factor D 24 2 Cleaves Factor B
Properdin 55 15-25 Stabilizes C3bBb (C3 convertase)
Lectin Pathway Proteins
MBL 200-600 0.0002-10 Binds to mannose
MASP-1 93 1.5-12 Unknown
MASP-2 76 Unknown Cleaves C4 and C2
CLASSICAL PATHWAY
• It is the main antibody-directed mechanism for
triggering complement activation, however not
all immunoglobulins are able to activate this
pathway. Those classes that can activate CP
includes IgM, IgG1, IgG2 and IgG3.
• IgM is the most efficient because it has multiple
binding site, thus needing only one molecule to
initiate the cascade.
• Two IgG molecules must attach to antigen
before they can bind a complement.
• In addition to antibody, few substances that can
bind complement directly to initiate the
classical pathway includes CRP, several viruses,
mycoplasmas, some protozoa and certain gram-
negative bacteria such as [Link].
• Divided into 3 main stages: recognition unit,
activation unit, and membrane attack complex.
Recognition Unit
• The first complement to be bind is the C1 complex
which is tightly bound by calcium. It consists of 3
subunits: C1q, C1r, C1s.
• C1q recognizes the Fc region of the two adjacent
antibody molecule that is attached on the surface
of the target cell. At least 2 globular heads of the
C1q must be bound to initiate the classical
pathway.
• Binds at the CH3 region of IgM
• Binds at the CH2 region of IgG
• Upon binding of C1q to antibody molecules, it will
activate the inactive C1r and C1s to become
active, converting them into active serine
protease.
• Active C1r cleaves thioester bond of C1s
• Active C1s cleaves C4 and C2
• Once C1s is activated, the recognition stage ends.
Activation Unit
• The activation unit results in the production of the
C5 convertase.
• Active C1s cleaves C4 to split off into C4a and C4b.
C4b must bind with protein, in this case, the C2 or
else it will be hydrolyzed to form the iC4b
(inactivated C4b). C4b also attracts at least other
30 C4 molecules to be cleaved by C1s. C4b will
combined with C2 to form “C4b2” and is stabilized
by magnesium. C4a serves as an anaphylatoxin.
• Active C1s also cleaves C2 on “C4b2” to split off
into C2a and C2b. This is the only complement
that has an “a” designation given with an enzyme
activity.
• Shedding of the C2b forms the “C4b2a” known as
the C3 convertase. It is an active enzyme that has
a half-life of 15 seconds to 3 minutes, so it must
bound quickly to C3.
Activation Unit
• C3 is the major complement of the body. It serves
as a pivotal point of all 3 pathways.
• C3 convertase (C4b2a) cleaves C3 to form C3a and
C3b. C3 convertase can cleave about 200
molecules of peripheral C3. C3a serves as an
anaphylatoxin, while unbound C3b serves as a
powerful opsonin.
• C3b must be bound to C4b2a (C3 convertase) to
form the new enzyme “C4b2a3b” or also known
as the C5 convertase. Formation of C5 convertase
depicts the last step of activation unit.
Membrane Attack Complex
(MAC)
• C5 convertase cleaves C5 to form C5a and C5b. C5a is a
strong chemoattractant and involved in recruitment of
effector cells.
• Subsequent binding involves C6, C7, C8 and C9, forming
the C5b6789 complex.
• C5b binds with C6 to be stabilized to form “C5b6” and
attaches to the another site on cell membrane. This must
be attached quickly with C6 because it is rapidly
inactivated due to it being a highly labile.
• C5b6 binds with C7 forming the C5b67 which has an high
affinity to lipid constituents of membrane.
• C5b67 binds with C8 forming the C5b678 which forms a
small hole in the membrane.
• C5b678, particularly the C8 complement binds with 1-18
molecules of C9 forming the C5b6789. C9 polymerizes to
form a hollow cylinder which constitutes the
transmembrane channel.
• Destruction of cell occurs through influx of water and
corresponding loss of electrolytes.
ALTERNATIVE PATHWAY
• Alternative pathway is important in such it is the
first pathway to be activated before full
development of the innate immunity.
• It is also known as the antibody-independent
pathway.
• Many unrelated triggering substances for
alternative pathway includes: bacterial
lipopolysaccharides, fungal cell walls, yeasts,
viruses, virally infected cells, tumor cells, and
some parasites.
• C3 molecules are spontaneously hydrolyzed by
water to form C3b and C3a.
• C3b binds with Factor B to form C3bB and is
cleaved by Factor D to form C3bBb and Ba. Ba
serves as an inhibitor of the B cells.
ALTERNATIVE PATHWAY
• C3bBb is also known as the C3 convertase of
the alternative pathway. Presence of C3bBb
helps to cleave more C3 molecules which leads
to amplification of the activation.
• C3bBb degrades rapidly and must be stabilized
by properdin (C3bBbP) to increase the half-life
until such it is bound with other C3b fragments.
• C3bBb can cleave C5 but it is more efficient in
cleaving C3.
• If some of the produced C3b binds to the C3bBb
and stabilized by properdin, it will form
C3bBb3bP which has a high affinity with C5 and
exhibits C5 convertase activity of the alternative
pathway.
• C5 is cleaved to produced C5b and from this
point on, both the alternative and the classical
pathways are identical.
LECTIN PATHWAY
• It is also known as the mannose-binding or
mannan-binding lectin.
• It represents another means of activating
complement without antibodies. Lectins are
proteins that bind to carbohydrates.
• MBL acts similar to C1q and associated with
serine proteases MASP-1, 2 and 3.
• Once MBL binds to mannose or related
carbohydrates present on the target cell
surface, MASP-2 which is homologous to
C1s, acts as role of cleaving C4 and C2.
• Once C4 and C2 are cleaved, the rest of the
pathway is similar to the classical pathway.
POINTS TO REMEMBER
Action Examples
Opsonization C3b and C1q – enhances phagocytosis
Chemoattractant C5a and C5678 complex – attracts PMNs
C5a – enhances adhesiveness of neutrophils,
activates integrins.
Anaphylatoxin C3a, C4a and C5a
Causes degranulation of mast cells
Causes bronchoconstriction
REGULATORY SYSTEM OF COMPLEMENT
ACTIVATION
• These molecules inhibits the complement pathways to prevent greater tissue damage due to
proinflammatory effects of the complement activation.
Regulators of the Complement Pathways
Serum protein MW(kD) Conc. (mg/mL) Function
C1 inhibitor (C1INH) 105 240 Dissociates C1r and C1s from C1q
- Classical inhibitor Inactivates MASP-2
Factor I 88 35 Cleaves C3b to iC3b and C3f; iC3b is further broken down into C3c and C3dg in
- All pathways conjunction with cofactor CR1
Cleaves C4b, inactivating the C5 convertase.
Factor H 150 300-450 Cofactor with Factor I to inactivate C3b.
- All pathways Prevents binding of Factor B to C3b
C4-binding protein (C4BP) 520 250 Acts a cofactor with Factor I to inactivate C4b
- All pathways
S protein (vitronectin) 84 500 Prevents attachment of C5b67 complex to cell membrane.
- All pathways
Receptors on the Cell Membrane for Complement Components
Receptor Ligand Cell Type Function
CR1 (CD35) C3b, iC3b, C4b RBC, PMNs, monocyte, macrophage, eosinophils, B Cofactor for factor I, mediates transport of
and T cells, follicular dendritic cells immune complexes
CR2 (CD21) C3dg, C3d, iC3b B cells, follicular dendritic cells, epithelial cells B cell coreceptor for antigen with CD19
CR3 (CD11b/CD18) iC3b, C3d, C3b monocyte, macrophage, PMNs, NK cells Adhesion and increased activity of
phagocytic cells
CR4 (CD11c/CD18) iC3b, C3b monocyte, macrophage, PMNs, NK cells, activated Adhesion and increased activity of
T and B cells, dendritic cells phagocytic cells
DAF (CD55) C3b, C4b RBC, PMNs, platelets, monocytes, endothelial cells, Dissociates C2b or Bb from binding sites,
fibroblasts, T cells, B cells, epithelial cells thus preventing formation of C3 convertase.
Protects the cell from bystander lysis
MIRL (CD59) C8 RBC, PMNs, platelets, monocytes, endothelial cells, Prevents insertion of C9 into the cell
epithelial cells membrane
MCP (CD46) C3b, C4b monocyte, macrophage, PMNs, platelets, T cells, B Cofactor for factor I cleavage of C3b and
cells, endothelial cells C4b.
DAF – decay accelerating factor; MIRL – membrane inhibitor of reactive lysis; MCP – membrane cofactor protein
COMPLEMENT DEFICIENCIES
Deficient Associated Disease
Component
• Hereditary deficiencies of any complement
protein, except for C9 usually manifests itself C1 (qrs) Lupuslike syndrome; recurrent infection
with an increased susceptibility to infections. C2 Lupuslike syndrome; recurrent infections
• C2 deficiency is the most common, found in C3 Severe recurrent infections; glomerulonephritis
1:20000 individuals.
C4 Lupuslike syndrome
• C3 deficiency is the most serious of all
because it is the key mediator of all C5-8 Neisseria infections
pathways. C9 No known disease
• Individuals with PNH have a deficiency in DAF C1INH Hereditary angioedema
on their RBCs, hence, the cells are subjects to
bystander lysis once complement system has DAF Paroxysmal nocturnal hemoglobinuria
been triggered.
MIRL Paroxysmal nocturnal hemoglobinuria
• Hereditary angioedema is due to C1INH
deficiency. Lac of C1INH leads to continuous Factor H or I Recurrent pyogenic infection
cleavage of C4 and C2 and creating C4a and MBL Pneumococcal diseases, sepsis, Neisseria
C2b which increases the vascular infections
permeability leading to edema.
Properdin Neisseria infections
MASP-2 Pneumococcal diseases