IMMUNOLOGY AND SEROLOGY

CHAPTER 5: The Soluble Mediators of Innate System

Ref: Turgeon

THE COMPLEMENT SYSTEM

 Complement is a heat-labile series of 18 plasma

proteins, many of which are enzymes or proteinases
 Are a major fraction of the beta-1 and beta-2
globulins

Naming:

 named with a capital C followed by a number

 small letter after the number Activation of Complement
o indicates that the protein is a smaller protein
 Complement components are present in the
resulting from the cleavage of a larger circulation in an inactive form
precursor by a protease
 Control proteins are normally present to inhibit
o E.g. C3a, C3b
uncontrolled complement activation:
 the larger fragment is designated as “b” and the o C1 INH,
smaller fragment as “a.” o Factor I
o EXCEPT:
o Factor H
 designation of the C2 fragments;
o C4-binding protein (C4-bp)
the larger fragment is designated
C2a and the smaller fragment is Activation of Classic Pathway
C2b.
 Initiated by the bonding of the C1 complex to
Three Pathways: antibodies bound to an antigen on the surface of a
bacterial cell; consisting of:
1. Classic Pathway
o C1q
2. Alternative Pathway
o Proteins are called Factors; symbolized by o C1r
letters such as B o C1s
3. Mannose-binding lectin pathway Activation of Alternative Pathway

 The three pathways converge at the point of cleavage  Initiated by contact with a foreign surface such as the
of C3 to C3b which in turn leads to the activation of polysaccharide coating of a microorganism and the
lytic complement sequences C5 through C9 and cell covalent binding of a small amount of C3b to
destruction hydroxyl groups on cell surface carbohydrates and
proteins.
 The pathway is activated by low-grade cleavage of
C3 in plasma.
.
Activation of Mannose-Binding Lectin Pathway

 Initiated by binding of the complex of mannose-

binding lectin and associated serine proteases
(MASP1 and MASP2) to arrays of mannose groups
on the surface of a bacterial cell

Enzyme Activation

 After complement is initially activated, each enzyme

precursor is activated by the previous complement
component or complex, which is a highly specialized
proteinase.
 This converts the enzyme precursor to its
catalytically active form by limited proteolysis
 The pathways leading to the cleavage of C3 are
triggered enzyme cascades.
 During this activation process, a small peptide
fragment is cleaved, a membrane-binding site is
exposed, and the major fragment binds.
 As a consequence, the next active enzyme of the
sequence is formed.
 Because each enzyme can activate many enzyme
precursors, each step is amplified until the C3 stage;
 Complement Receptors

 Complement receptor 1 (CR1) is important in:

o enhancing phagocytosis
o Plasmodium falciparum adhesin PfRh4
 binds to CR1 on human
erythrocytes
o CR1 is a complement regulator and
o immune adherence receptor on erythrocytes
required for shuttling C3bC4b-opsonized
particles to the liver and spleen for
phagocytosis.
 CR3 is important in:
o host defense mechanisms.

Effects of Complement Activation

 Physiologic consequences include:

o Blood vessel dilation
o Increased vascular permeability

 Cellular consequences:
o Cell activation, such as production of
inflammatory mediators.
o Cytolysis or hemolysis, if the cells are
erythrocytes.
 The most important biologic role of
complement in blood: production
of cell membrane lysis of
antibody-coated targets.
o Opsonization, which renders cells
vulnerable to phagocytosis.

Activation of complement may also:

 Physiologic concentrations of complement may

induce profound alterations in the molecular
weight, composition, and solubility of immune
complexes

 Mediating hypersensitivity reactions

o may occur from direct alternative pathway
activation by immunoglobulin E (IgE) or
o through a sequence initiated by the activated
Hageman coagulation factor that causes the
generation of plasmin,
 activates the classic pathway.
o Both leads to the generation of
anaphylatoxins

Classical Pathway

 one of the major effector mechanisms of antibody-

mediated immunity
 Component: C1-C9
 The sequence of component activation—C1, 4, 2, 3,
5, 6, 7, 8, and 9
 C3 – present in plasma in the largest quantities
 o Fixation of C3 – major quantitative reaction 4. This exposes a reactive binding site on the larger
of the complement cascade fragment, C3b. Consequently, clusters of C3b
 components are made in hepatic parenchymal cells, molecules are activated and bound near the C4bC2a
o EXCEPT: complex.
 C1 5. Each catalytic site can bind several hundred C3b
 (a calcium-dependent complex of molecules, even though the reaction is very efficient
the three glycoproteins C1q, C1r, because C3 is present in high concentration
and C1s), 6. Only one C3b molecule combines with C4bC2a to
 Synthesized in the epithelium of form the final proteolytic complex of the complement
the gastrointestinal and urogenital cascade.
tracts

The classic pathway has three major stages:

1. Recognition
Membrane Attack Complex
2. Amplification of proteolytic complement cascade
3. Membrane attack complex (MAC)  unique system that builds up a lipophilic complex in
cell membranes from several plasma proteins.
Recognition
1. To initiate C5b fixation and the MAC, C3b splits C5a
 C1 complex (C1q, C1r, C1s)– recognition unit from the alpha chain of C5
o an interlocking enzyme system 2. When fully assembled in the correct proportions, C7,
C6, C5b, and C8 form the MAC
Steps: o The C5bC6 complex is hydrophilic
 addition of C7; it has additional
1. Initiation of the pathway triggered by recognition by
detergent and phospholipid-binding
complement factor C1 of antigen-antibody complexes
properties as well.
on the cell surface.
3. Once membrane bound, C5bC6C7 is relatively stable
2. When C1 complex interacts with aggregates of
and can interact with C8 and C9.
immunoglobulin G (IgG) with antigen on a cell’s
4. The C5bC6C7C8 complex polymerizes C9 to form a
surface, , C1r and C1s, are activated
tubule (pore), which spans the membrane of the cell
being attacked, allowing ions to flow freely between
 A single IgM molecule is potentially able to fix C1,
the cellular interior and exterior.
but at least two IgG molecules are required for this
5. By complexing with C9, the osmotic cytolytic
purpose.
reaction
 The amount of C1 fixed is directly proportional to the is accelerated.
concentration of IgM antibodies, although this is not
o This tubule is a hollow cylinder with one
true of IgG molecules.
end inserted into the lipid bilayer and the
other projecting from the membrane.
3. C1s is weakly proteolytic for free intact C2, but is
highly active against C2 that has complexed with
 No further proteinases are generated in the classic
C4b molecules in the presence of magnesium (Mg2+)
complement sequence
ions
4. This reaction will occur only if the C4bC2 complex
 Other bound C3b molecules not involved in the
forms close to the C1s
C4b2a3b complex form an opsonic macromolecular
5. The resultant C2a fragment joins with C4b to form
coat on the erythrocyte or other target,
the new C4bC2a enzyme, or classic pathway C3
o which renders it susceptible to immune
convertase.
6. The catalytic site of the C4bC2a complex is probably adherence by C3b receptors on phagocytic
in the C2a peptide. cells
7. A smaller C2b fragment from the C2 component is
lost to the surrounding environment  The presence of hydrophobic and hydrophilic groups
within the same complex may account for its
Amplification of Proteolytic Complement Cascade tendency to polymerize and form small protein
micelles (a packet of chain molecules in parallel
1. Once C1s is activated, the proteolytic complement
arrangement).
cascade is amplified on the cell membrane through
o can attach to any lipid bilayer within its
sequential cleavage of complement factors and
radius,
recruitment of new factors until a cell surface
 produces the phenomenon of
complex containing C5b, C6, C7, and C8 is formed.
reactive lysis on innocent so-called
2. The complement cascade reaches its full amplitude at
bystander cells.
the C3 stage, which represents the heart of the system
3. The C4bC2a complex, the classic pathway C3 ALTERNATIVE PATHWAY
convertase, activates C3 molecules by splitting the
peptide, C3 anaphylatoxin, from the N-terminal end  predominantly a non–antibody-initiated pathway
of the peptide of C3.
  Microbial and mammalian cell surfaces can activate
the alternative pathway in the absence of specific 2. Biological effects of proteolytic fragments of
antigen-antibody complexes. complement
 Factors capable of activating alternative pathway: o active fragments mediate their effects by
o Inulin binding to specific receptors expressed on
o Zymosan various types of cells, including phagocytic
o Bacterial polysaccharides leukocytes and the endothelium
o Endotoxins
o Aggregated IgG2, IgA, and IgE Alterations in Complement Levels
Paroxysmal nocturnal hemoglobinuria (PNH)  Complement activation is also associated with
 patient’s erythrocytes act as an activator and result in intravascular thrombosis, which leads to ischemic
excessive lysis of these erythrocytes injury to tissues.
 Complement levels may be abnormal in certain
Note: disease states
o Ex. rheumatoid arthritis, systemic lupus
 C1, C4, and C2—do not participate in the cascade erythematosus [SLE]
sequence. o and in some genetic disorders.
 C3a component is the counterpart of C2a in the
classic pathway.
 C2 of the classic pathway structurally resembles
factor B of the alternative pathway

 The omission of C1, C4, and C2 is possible because

activators of the alternative pathway catalyze the
conversion of another series of normal serum
proteins, which leads to the activation of C3.

1. The uptake of factor B onto C3b occurs when C3b is

bound to an activator surface.
o C3b in the fluid phase or attached to a
nonactivator surface will preferentially bind
to and therefore prevent C3b,B formation.

2. C3b and factor B combine to form C3b,B, which is

converted into an active C3 convertase, C3b,Bb.
o results from the loss of a small fragment, Ba
Elevated Complement Levels
(glycine-rich α2-globulin believed to be
physiologically inert), through the action of  inflammatory conditions
the enzyme, factor D.  trauma
 acute illness such as myocardial infarction
3. The C3b,Bb complex is able to convert more C3 to o because separate complement component
C3b, which binds more factor B and the feedback are acute-phase proteins
cycle continues.  Ex. C3
Factor H  common and nonspecific
 limited clinical significance
 major controlling event of the alternative pathway
Decreased Complement Levels
 which prevents the association between C3b and
factor B  Complement has been excessively activated recently.
 Factor H blocks the formation of C3b,Bb, the  Complement is currently being consumed.
catalytically active C3 convertase of the feedback  A single complement component is absent because of
loop. a genetic defect
 competes with factor B for its combining site on C3b,
eventually leading to C3 inactivation Hypocomplementemia

The association of numerous C3b units, factor Bb, and  can result from the complexing of IgG or IgM
properdin on the surface of an aggregate of protein or the antibodies capable of activating complement
surface of a microorganism has potent activity as a C5
convertase The following three types of complement deficiency can

cause increased susceptibility to pyogenic infections:

BIOLOGICAL FUNCTIONS OF COMPLEMENT 1. Deficiency of the opsonic activities of complement

PROTEINS 2. Any deficiency that compromises the lytic activity of
complement
3. Deficient function of the mannose-binding lectin
pathway
1. Cell lysis by the membrane attack complex (MAC)
o leads to osmotic lysis of a cell DIAGNOSTIC EVALUATION
 C1 Esterase Inhibitor (C1 Inhibitor)
o Deficiency: hereditary angioedema
 C1r, C1s, C2, C3, C4, C5, C6, C7, C8
o Autoimmune disease (especially SLE)
o Arthritis
o chronic glomerulonephritis
o infections, and vasculitis.
 C1q
o Deficiency:
 hypocomplementemic urticarial
vasculitis
 severe combined
immunodeficiency (SCID)
 X-linked hypogammaglobulinemia