Complement

Immunology and Serology

BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
The Complement Functions of the Complement

● A group of serum proteins involved Opsonization

in the control of inflammation, the
activation of phagocytes and the lytic 1. Activation of Leukocytes
attack on cell membranes 2. Lysis of target cells
● The system can be activated by
interaction with the immune system
● The complement system is a Opsonization
biochemical cascade that helps, or
“complements”, the ability of
antibodies to clear pathogens from an
organism.
● The complement system consists of a
number of small proteins found in the
blood, generally synthesized by the
liver, and normally circulating as
inactive precursors or pro-proteins.
● When stimulated by one of several
triggers, proteases in the system
cleave specific proteins to release
cytokines and initiate an amplifying
cascade of further cleavages.
● The end-result of this activation
● Involves coating of the target with
cascade is massive amplification of
certain complement proteins
the response and activation of the
● Phagocytic cells carrying receptors for
cell-killing membrane attack complex.
these complement proteins bind and
● Over 25 proteins and protein
endocytose the opsonized particles
fragments make up the complement
system, including serum proteins,
serosal proteins, and cell membrane Activation of Leukocytes
receptors.

● Follows from the interaction of

Three major biochemical pathways complement proteins with specific cell
activate the complement system surface receptors
● Activation of the complement results in
1. Classical Complement Pathway the cleavage of complement proteins
2. Alternative Complement Pathway to yield small fragments which can
3. Mannose Binding Lectin Pathway diffuse readily
● There are specific receptors on PMNs
● All three pathways merge at the & macrophages that causes directed
cleavage of C3, which leads to the cellular movement (chemotaxis) and
generation of three effector activation
mechanisms: recruitment of ● Similar receptors on lymphocytes and
inflammatory cells, opsonization, and antigen-presenting cells bind
the generation of the membrane attack complement-opsonized immune
complex complexes and enhance specific
immune responses

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Complement
Immunology and Serology
BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
● C4b and C2a bind to form the classical
pathway C3-convertase (C4b2a
complex), which promotes cleavage of
C3 into C3a and C3b; C3b later joins
with C4b2a (the C3 convertase) to
make C5 convertase (C4b2a3b
complex). The inhibition of C1r and
C1s is controlled by C1-inhibitor.

Lysis

● The third physiological consequence

of complement, lysis is caused by the
insertion of a hydrophobic plug into
lipid membrane bilayers, allowing
osmotic disruption of the target.

The Classical Pathway The Alternative Pathway

● The classical pathway is triggered by ● The alternative pathway is triggered by

activation of the C1-complex spontaneous C3 hydrolysis directly
(composed of 1 molecule of C1q, 2 due to the breakdown of the thioester
molecules of C1r and 2 molecules of bond via condensation reaction (C3 is
C1s, thus forming C1qr2s2), which mildly unstable in aqueous
occurs when C1q binds to IgM or IgG environment) to form C3a and C3b.
complexed with antigens (a single IgM ● C3b is then capable of covalently
can initiate the pathway, while multiple binding to a pathogenic membrane
IgGs are needed), or when C1q binds surface if it is near enough. If there is
directly to the surface of the pathogen. no pathogen in the blood, the C3a and
● Such binding leads to conformational C3b protein fragments will be
changes in the C1q molecule, which deactivated by rejoining with each
leads to the activation of two C1r (a other.
serine protease) molecules. ● Upon binding with a cellular
● They then cleave C1s (another serine membrane C3b is bound by factor B to
protease). The C1r2s2 component form C3bB.
now splits C4 and then C2, producing ● This complex in presence of factor D
C4a,C4b,C2a,and C2b. will be cleaved into Ba and Bb.

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Complement
Immunology and Serology
BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
● Bb will remain covalently bonded to ● Anaphylotoxins to basophils causes
C3b to form C3bBb, which is the vessel dilation, leading to an increased
alternative pathway C3-convertase. blood flow to an inflammatory site
● The protein C3 is produced in the liver. ● C4a is similar to C3a, but C4a is far
● The C3bBb complex, which is less effective in its biological effects on
"hooked" onto the surface of the a molar basis
pathogen, will then act like a "chain ● C5a is the most potent of all
saw," catalyzing the hydrolysis of C3 anaphylotoxins
in the blood into C3a and C3b, which
positively affects the number of C3bBb
hooked onto a pathogen. MBL Pathway
● After hydrolysis of C3, C3b complexes
to become C3bBb3b, which cleaves ● The MBL pathway is activated either
C5 into C5a and C5b. C5b with C6, through recognition of carbohydrates
C7, C8, and C9 (C5b6789) complex to by MBL or by recognition of
form the membrane attack complex, N-acetylated compounds by ficolins.
also known as MAC, which is inserted ● Upon binding, MASPs become
into the cell membrane, "punches a activated and cleave C4 and C2.
hole," and initiates cells lysis.
● C5a and C3a are known to trigger **MASP:mannose-binding-lectin-associated
mast cell degranulation. serine proteases.

Biologic Functions Associated with Inherited Deficiencies in Complement and

Activation Complement-related Proteins

● Anaphylotoxins C3a, C4a and C5a are

Deficient Component / Reported major clinical
biologically active, low-molecular Subcomponent correlates
weight peptides that are generated by
the enzymatic cleavage of their parent C1q Glomerulonephritis, SLE
molecules during complement
activation C1r Syndrome resembling SLE

● Anaphylotoxins are defined by their

C1s SLE
biological effects on smooth muscle,
mast cells and small blood vessels C4 Syndrome resembling SLE
and peripheral leukocytes
C2 SLE, discoid LE, juvenile
Specific effects include: rheumatoid arthritis,
glomerulonephritis

1. Degranulation of mast cells and C3 Recurrent pyogenic

basophils with subsequent release of infections, glomerulonephritis
various mediators in the absence of
cytotoxicity C5 Recurrent disseminated
neisserial infections, SLE
2. Induction of human neutrophil
aggregation C6 Recurrent disseminated
3. Smooth muscle contraction neisserial infections
4. Enhanced vascular permeability
5. Induction of serotonin release C7 Recurrent disseminated
neisserial infections,
(demonstrated in guinea pigs) Raynaud’s phenomenon
6. Thromboxane release from guinea
pigs macrophage C8 Recurrent disseminated
7. Stimulation of mucus release from neisserial infections
goblet cells
C9 None Identified

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Complement
Immunology and Serology
BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
● The test sample is then added and the
Regulation of Activation
degree of hemolysis is related to the
presence of later-acting components
● Once the hydrophobic C5b67 complex ● Immunoassays – immunochemical
has formed, it can insert itself into assays are generally simpler to
other cell membranes close to the perform than those for evaluating
primary surface on which complement functional activity
activation is focused ● These are highly specific and requires
● This is the process of reactive lysis fewer specialized reagents and
which if unregulated could have considerably less time
damaging consequences to self/host ● Reagents are commercially available,
tissues either serum or plasma can be used,
● Certain proteins inhibit this process by and the commonly available methods
binding to fluid phase C5b67 before it of freezer storage are sufficient.
can attach to self membranes
● S Protein (vitronectin) usually in the
plasma is the most abundant In summary…
● It forms SC5b67 complex which is
unable to insert into a lipid bilayers Complement refers to 14 distinct serum
● If C8 binds to C5b67 in the fluid proteins, 9 components
phase, this also forms a complex
incapable of membrane insertion as 1. Complement plays a role in the
does the binding of LDL cytolytic destruction of cellular
● Host cells also bear membrane antigens by specific Ig
proteins that protect them against lysis 2. Not all cellular Ag are susceptible to
by the membrane attack complex complement-mediated cytolysis
● 2 proteins has been identified to 3. IgM and IgG react with compoment,
mediate process of restriction except with some IgG subclasses
● First is CD59, is a protein anchored by 4. Complement is bound to all Ag-Ab
a glycophospholipid foot. It is widely complex
distributed in cell membranes and 5. Complement is found in sera of
inhibits the insertion and mammals, lower animals, birds,
polymerization of C9 into cell fish, amphibia and sharks
membranes bearing C5b-8 6. Complement from one species will
● Second is the homologous restriction usually react with immunoglobulins
protein that has similar activity and from another species from the same
distribution to CD59 but is a weaker taxonomic order. This interaction
inhibitor of C9 insertion decreases as the taxonomic
position of the 2 becomes more
distant
Measurement of Individual Complement 7. Complement contributes to
Components chemotaxis, opsonization, immune
adherence, anaphylotoxin
formation, virus neutralization, and
● Functional assays are considered to other physiologic functions
be both sensitive and precise tools for 8. Complement can be activated by
measuring the activity of complement nonserologic reactions
component
● The level of activity is measured by The Component Nomenclature
using pure preparations of each
component added sequentially to The complement components are
antibody coated erythrocytes until the numbered C1 to C9
step is reached in the activation
sequence just prior to addition of the 1. The individual peptide chains of
component to be measured these proteins are designated by
Greek letters

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Complement
Immunology and Serology
BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
2. Fragments are designated by lower ● C3 is the 3rd activation unit
case arabic letters (C3a) ● The most abundant complement
3. The letter i is added to indicate component in serum
inactivation (C3bi or iC3b) ● C3 is split by C3 convertase into
4. Activation is sometimes indicated C3a & C3b
by a horizontal bar over the ● C3b attaches to activated C4b2a
designation. This is more commonly ● C4b2a3b is formed, the C5
used to indicate enzymatic activity convertase
5. At least 5 additional proteins
participate in the alternate pathway ● C5: 1st membrane attack unit
6. Several proteins that modulate ● C5 convertase cleaves into 2
complement derived activities have subunits – C5a & C5b
been isolated ● C5a is released into the body fluids
where as anaphylatoxin II, it acts as
Complement Activation: Classical a mediator of inflammation and a
Pathway chemotoxin for granulocytes
● C5b can be degraded to C5c & C5d

Complement activation (classical pathway ● C6 and C7: the 2nd & 3rd
begins with C1 activation. Membrane attack units
● C5b cleaves C6 into C6a and C6b
1. From this point, C4, C2, C3, and C5 ● Both C6b and C7 appear to bind to
to C9 participate in that order C5b by absorption
● C6 & C7 associate easily w/ C5 in a
● C1 is the recognition unit trimolecular complex
● It has 3 subunits: C1q, C1r, C1s
● C1q must attach to 2 Fc fragments ● C8 & C9: final membrane attack
to initiate the sequence units
● Exact mechanism for C1r and C1s ● C8 is inserted into the cell
activation remains membrane and disrupts it
irreversibly
● C4 is the activation unit ● C9 appears to enhance the activity
● C1s activates C4, w/c releases C4a of C8
and C4b
● C4b attaches to receptors on
erythrocyte surfaces, bacterial cell
membranes and other antigens. It
doesn’t attach to Ag-Ab complex
● C4a plays no further part in the
complement sequence

● C2 is the 2nd activation unit

● C2 is activated by C1s, though
probably only after the C1s-C4
interaction has taken place
● C2 occurs in small quantities in
serum
● C2 binds w/ C4b and is cleaved by
C1s into C2a & C2b
● C2b is labile and decays rapidly
● C2a binds to C4b in the presence of
magnesium to form C4b2a, a
proteolytic enzyme known as C3
convertase

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Complement
Immunology and Serology
BSMT-3 | SIR D. REYES | TRANSCRIBED BY: KRIZABEL 💉👩🏽‍⚕️
Complement Activation: The Alternative
Pathway

1. Many complex polysaccharides will

activate the alternative pathway
2. Serum proteins function in the
alternative pathway – factor B, factor
D, & properdin
3. The alternative pathway proceeds to
the C3 activator stage w/out the
participation or consumption of C1, C4
or C2
4. Once C3 has been split to form C3a,
the activation sequence proceeds to
C9 in the same way as the classical
pathway