Chapter 7
Complement System
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Chapter Overview
• Pathways of the complement system
• System controls
• Complement receptors and their biological roles
• Biological manifestations of complement
activation
• Complement and disease states
• Complement deficiencies and laboratory
detection of abnormalities
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Complement
• Complement is a series of more than 30 soluble
and cell-bound proteins that interact to enhance
host defense mechanisms against foreign cells
• Most plasma complement proteins are
synthesized in the liver
• Exceptions:
• C1 components (produced by intestinal epithelial
cells)
• Factor D (made in adipose tissue)
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Complement
• White blood cells are additional sources of
early complement components:
• C1, C2, C3, C4
• Most of these proteins are zymogens that are
converted to active enzymes in a very precise
order
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Complement Fragments/Components
• Act as opsonins
• Specific receptors are present on phagocytic cells
• Metabolism and clearance of immune complexes
are enhanced
• Increase vascular permeability
• Recruit monocytes and neutrophils to the area
of antigen concentration
• Trigger secretion of immunoregulatory
molecules that amplify the immune response
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Complement System Activation
• Three pathways:
• Classical pathway
• Lectin pathway
• Alternative pathway
• Multiple pathways can be involved
• End product: Lysis of the invading cell
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The Classical Pathway
• Has three stages
• Activated by IgM,
IgG1, IgG2, and
IgG3
• IgM: Most efficient
due to multiple
binding sites
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The Classical Pathway
• Also activated by substances that bind
complement directly:
• C-reactive protein
• Certain viruses
• Mycoplasmas
• Certain gram-negative bacteria
• Some protozoa
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The Classical Pathway: Recognition Unit
• C1
• First complement
component to bind
• Consists of three
subunits:
• One Clq
• Two Clr
• Two Cls
• Stabilized by
calcium
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The Classical Pathway: Recognition Unit
• Clq “recognizes” the Fc region of two adjacent
antibody molecules
• At least two of the globular heads of Clq must
be bound to initiate the classical pathway
• The zymogens Clr and Cls are converted into
active enzymes as binding of Clq occurs
• Autoactivation of Clr results from a change that
takes place as Clq is bound
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The Classical Pathway: Recognition Unit
• When activated, Clr cleaves a thioester bond
on Cls, which activates it
• Cls has a limited specificity, with its only
substrates being C4 and C2
• When Cls is activated, the recognition stage
ends
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The Classical Pathway: The Activation Unit
• Begins when Cls
cleaves C4 to
create C4b and C2
to create C2a
• C3 convertase is
formed
• Ends with
production of C5
convertase
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The Classical Pathway: The Activation Unit
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The Classical Pathway: Membrane Attack Complex
• The process is
initiated when C5
convertase splits
C5 into C5a and
C4b
• C5b attaches to the
cell membrane to
initiate formation
of the MAC
(C5b6789)
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The Lectin Pathway
• Activates complement by recognizing
carbohydrates on microbial cell walls
• Serves as a defense mechanism in infancy in
between loss of maternal antibody and
development of full antibody response
• Involves three classes of recognition
molecules (lectins, ficolins, CL-L1)
• After cleaving of C4 and C2, is identical to the
classical pathway
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The Lectin Pathway
• Mannan-binding lectin (MBL)
• An acute-phase protein
• Binds to mannose or related sugars in a calcium-
dependent manner to initiate pathway
• MBL-associated serine proteases (MASPs)
• MASP-1, MASP-2, MASP-3
• Play enzymatic roles
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The Alternative Pathway
• Acts mainly as an
amplification loop
for classical or
lectin pathways
• Can be activated
on its own by
bacterial or fungal
cell walls, viruses,
tumor cells, or
some parasites
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Convergence of Pathways
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System Controls in Plasma
SERUM MOLECULAR CONCEN- FUNCTION INVOLVED
PROTEIN WEIGHT (KD) TRATION PATHWAYS
(MG/DL)
C1-INH 105 240 Dissociates C1r and C1s Classical
from C1q Lectin
Factor I 88 35 Cleaves C3b and C4b Classical
Lectin
Factor H 150 300–450 Cofactor with I to inactivate Alternative
C3b; prevents binding of B
to C3b
C4BP 520 250 Acts as a cofactor with I to Classical
inactivate C4b Lectin
S protein 84 500 Prevents attachment of Classical
(vitronectin) C5b67 complex to cell Lectin
membranes Alternative
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System Controls
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Cellular Complement Controls
RECEPTOR LIGAND CELL TYPE FUNCTION
DAF (CD55) C3b, C4b RBCs, neutrophils, platelets, Dissociates C2b or Bb
monocytes, endothelial cells, from binding sites, thus
fibroblasts, T cells, B cells, preventing formation of
epithelial cells C3 convertase
MIRL (CD59) C8 RBCs, neutrophils, platelets, Prevents insertion of C9
monocytes, endothelial cells, into cell membrane
epithelial cells
MCP (CD46) C3b, C4b Neutrophils, monocytes, Cofactor for factor I
macrophages, platelets, T cleavage of C3b and C4b
cells, B cells, endothelial cells
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Biological Roles of Complement Receptors
RECEPTOR LIGAND CELL TYPE FUNCTION
CR1 (CD35) C3b, iC3b, C4b RBCs, neutrophils, Cofactor for factor I;
monocytes, macrophages, mediates transport of
eosinophils, B and T cells, immune complexes
follicular dendritic cells
CR2 (CD21) C3dg, C3d, iC3b B cells, follicular dendritic B-cell co-receptor for
cells, epithelial cells antigen with CD19
CR3 iC3b, C3d, C3b Monocytes, macrophages, Adhesion and in-
(CD11b/CD18) neutrophils, NK cells creased activity of
phagocytic cells
CR4 iC3b, C3b Monocytes, macrophages, Adhesion and
(CD11c/CD18) neutrophils, NK cells, increased
activated T and B cells, activity of phagocytic
dendritic cells cells
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Biological Manifestations of Complement Activation
• Amplifies inflammatory response
• Facilitates recognition and destruction of
antigen by phagocytic cells
• Has a role in uptake and presentation of
antigens so a specific immune response can
occur
• Facilitates B-cell activation
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Biological Manifestations of Complement Activation
• Links innate and adaptive immunity
• Amplifies the inflammatory response
• Effector molecules
• Anaphylatoxins
• Chemotaxins
• Opsonins
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Complement and Disease States
• Complement can be harmful if:
• It is activated systemically on a large scale, as in
gram-negative septicemia
• It is activated by tissue necrosis, as in myocardial
infarction
• Lysis of RBCs occurs
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Complement Deficiencies
DEFICIENT COMPONENT ASSOCIATED DISEASE
C1 (q, r, or s) Lupus-like syndrome; recurrent infections
C2 Lupus-like syndrome; recurrent infections; atherosclerosis
C3 Severe recurrent infections; glomerulonephritis
C4 Lupus-like syndrome
C5–C8 Neisseria infections
C9 No known disease association
C1-INH Hereditary angioedema
DAF Paroxysmal nocturnal hemoglobinuria
MIRL Paroxysmal nocturnal hemoglobinuria
Factor H or factor I Recurrent pyogenic infections
MBL Pneumococcal diseases, sepsis, Neisseria infections
Properdin Neisseria infections
MASP-2 Pneumococcal diseases
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Laboratory Detection of Complement Abnormalities
• Techniques involve:
• Measurement of components as antigen in
serum
• Measurement of functional activity
• Immunologic assays of individual components
• Radial immunodiffusion (RID)
• Nephelometry
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Laboratory Detection of Complement Abnormalities
• Classical pathway assays
• Measure lysis
• Include:
• Hemolytic titration (CH50) assay
• CH50 test for lysis of liposomes
• Radial hemolysis in agarose plates
• Alternative pathway assays
• AH50
• ELISA
• Test systems addressing all three pathways
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Interpretation of Laboratory Findings
IMPAIRED CLASSICAL PATHWAY LECTIN PATHWAY ALTERNATIVE
FUNCTION OR PATHWAY
DEFICIENCY
C1q, C1r, C1s Low Normal Normal
C4, C2 Low Low Normal
MBL, MASP2 Normal Low Normal
B, D, P Normal Normal Low
C3, C5, C6, C7, Low Low Low
C8, C9
C1-INH Low Low Low
Factor H and I Low Low Low
Improperly Low Low Low
handled sera
Adapted from Seelen MA, et al. An enzyme-linked immunosorbent assay-based method for functional analysis of the three pathways of the
complement system. In: Detrick B, Hamilton RG, and Folds JD, eds. Manual of Molecular and Clinical Laboratory Immunology. 7th ed.
Washington, DC: ASM Press; 2006:124.
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Summary
• The complement system is a series of more
than 30 soluble and cell-bound proteins that
interact with the innate and adaptive
immune systems to enhance host defenses
against infection.
• Complement activities include lysis of foreign
or damaged cells, opsonization, increase in
vascular permeability, and attraction of
monocytes and macrophages to areas where
needed.
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Summary
• The classical complement pathway is
triggered by IgG or IgM binding to the surface
of pathogens.
• Three distinct units are involved in the
classical pathway:
• Recognition unit, consisting of C1qrs
• Activation unit, consisting of C2, C4, and C3
• MAC, consisting of C5, C6, C7, C8, and C9
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Summary
• The lectin pathway is activated by
carbohydrates present in microbial cell walls.
• Molecules distinct to the lectin pathway
include mannose-binding lectin (MBL), MASP-
1, MASP-2, and MASP-3.
• The alternative pathway is triggered by
bacterial and fungal cell walls, yeast, viruses,
tumor cells, and certain parasites.
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Summary
• Plasma protein regulators allow for control of
the complement system.
• Soluble regulators include C1-INH, C4BP,
factor H, factor I, and S protein.
• Cell-bound regulators include CR1, MCP, and
DAF.
• Specific complement receptors on host cells
enhance phagocytosis and stimulate other
accessory cells.
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Summary
• Effector molecules play a major role in
recognition and presentation of antigens,
activation of B cells, and maintenance of
immunologic memory.
• Anaphylatoxins increase vascular permeability.
• Chemotaxins attract phagocytic cells to a specific
area.
• Opsonins coat damaged or foreign cells to
enhance phagocytosis.
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Summary
• Complement deficiencies can place an
individual at risk for certain infections.
• Laboratory assays for individual complement
components include radial immunodiffusion
and nephelometry.
• The hemolytic titration or CH50 assay
measures lysis.
• The AH50 assay measures the activity of the
alternative pathway.
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