COMPLEMENT SYSTEM

THE COMPLEMENT SYSTEM

• complement is a complex series of more
than 30 soluble and cell-bound proteins
that interact in a very specific way to
enhance host defense mechanisms
against foreign cells.
 THE COMPLEMENT SYSTEM
• Paul Ehrlich coined the term complement
because it complements the action of
antibody in destroying microorganisms
(1890s)

• Jules Bordet was awarded the Nobel

Prize in 1919 for his role in elucidating the
nature of complement.
 THE COMPLEMENT SYSTEM

• Desirable effects:
– OPSONIZATION
– Lysis of foreign
cells and immune
disorders
 THE COMPLEMENT SYSTEM
• Undesirable effects:
– Inflammation
– Tissue damage
 THE COMPLEMENT SYSTEM
• Complement proteins
– are mostly synthesized in the liver
• C1 components – intestinal epithelial cells
• Factor D – made in adipose tissue.
– Are able to:
• Increase vascular permeability
• Recruit monocytes and neutro in the area of
antigen concentration
• Trigger secretion of immunoregulatory molecules
that amplify immune response
 Three different pathways to
activate complement system
• The CLASSICAL PATHWAY, which involves nine
proteins that are triggered by antigen–antibody
combination.
• The ALTERNATIVE PATHWAY, was originally
called the properdin system, because the protein
properdin was thought to initiate this pathway.
• The LECTIN PATHWAY, is another antibody-
independent means of activating complement
proteins.
– Its major constituent, mannose- (or mannan-) binding
lectin (MBL), adheres to mannose found mainly in the
cell walls or outer coating of bacteria, viruses, yeast,
and protozoa.
 Functions of the Complement
System
• CELL LYSIS by the membrane attack
complex (MAC)

• Effector functions of complement

components
– C3b – Opsonin
– C5a, C3a, C4a – anaphylatoxins
– C5a – chemotaxin
 Nomenclature of Complement
Components
• The components of complement are
numbered C1 to C9
• Fragments are designated by lower – case
Arabic letters (i.e., C3a)
 The Classical Pathway
• The classical pathway or cascade is the
main antibody directed mechanism for
triggering complement activation
• IgM > IgG 3 >IgG1> IgG2
• C-reactive protein, several viruses, mycoplasmas,
some protozoa, and certain gram-negative bacteria
such as Escherichia coli.
 The Classical Pathway
• Recognition Unit
• Activation Unit
• Membrane Attack Complex
 The Classical Pathway
Recognition Unit – C1
• C1 is composed of 3
molecules held together by
CALCIUM (Ca2+)
• 3 subunits are called C1q, C1r
and C1s
• C1q must attach to two Fc
fragments to initiate the
sequence.
 The Classical Pathway
Activation
• Requires IgG or IgM
• IgM and IgG3 antibodies are most efficient
at complement activation
 The Classical Pathway
Activation Unit – C4
• C1s activates C4
• C4 releases C4a and C4b
• C4a plays no role in the complement
cascade
• C4b attaches to receptors on RBCs,
bacterial cell membranes and other
antigens
 The Classical Pathway

Second Activation Unit – C2

• C1s also activates C2 after its interaction with C4
• When combined with C4b, C2 releases C2a and C2b
• C2a binds to C4b in the presence of MAGNESIUM
(Mg2+) to form C4b2a (also known as C3
CONVERTASE)
• C2b is labile and decays rapidly
 The Classical Pathway
Third Activation Unit – C3
• C3 is the most abundant complement
component
• C3 convertase splits C3 into C3a and C3b
• C3b combines with C4b2a to form C4b2a3b
(also known as C5 CONVERTASE)
• Represents the most significant step in the
entire process of complement activation
• C3b also serves as a powerful opsonin
 The Classical Pathway
• The membrane attack complex (MAC) is
a unique system that builds up a lipophilic
complex in cell membranes from
several plasma proteins.
 The Classical Pathway
First Membrane Attack Unit – C5
• C5 convertase cleaves C5 into C5a and
C5b
• C5a is released into the body fluids where
it acts as ANAPHYLATOXIN and
CHEMOTAXIN
• C5b activates C6 and C7
 The Classical Pathway
Second and Third Membrane Attack
Units – C6 and C7
• C6 binds to C5b thereby stabilizing it
• This complex then attaches to the cell
surface
• C7 binds next, forming a trimolecular
complex which allows insertion of C7 part
of the C5b67 complex into the membrane
of the target cell
 The Classical Pathway
Final Membrane Attack Units – C8 and
C9
• C8 binds to C7 and forms a small hole in
the membrane of the target cell
• The pore formed by the binding of C5b678
is small, capable of lysing erythrocytes
but not nucleated cells.
• For lysis of nucleated cells to be
achieved, C9 must bind to the complex.
THE ALTERNATIVE PATHWAY
 The Alternative Pathway
• First described by Pillemer and his
associates in the early 1950s
• Does not require specific antibody for
activation

• Activation: POLYSACCHARIDES AND

LIPOPOLYSACCHARIDES on bacterial cell
walls, fungal cell walls, yeast, viruses,
virally – infected cells, tumor cell lines
and some parasites.
 The Alternative Pathway
• Bypasses (does not use) C1, C2, and C4
found in the classical pathway

• Factors unique in this pathway are:

– PROPERDIN (factor P)
– Factor B
– Factor D
 The Alternative Pathway
• Starts with the conversion of C3 into C3a and C3b.
– Water is able to hydrolyze a thioester bond and thus
spontaneously activates a small number of these
molecules.
• C3b (a.k.a. iC3) binds to Factor B
• Bound Factor B is cleaved by factor D into Ba and
Bb.
– Factor D has the lowest concentration of all the
complement proteins.
• Bb remains attached to C3b forming the initial C3
convertase of the alternative pathway
In plasma, native
C3 is not stable
 The Alternative Pathway
• Properdin binds to the C3bBb (C3
convertase) to STABILIZE IT
• C3 convertase cleaves C3 into C3a and C3b
• C3b joins C3bBb to form C3bBb3b (also
known as C5 convertase of the alternative
pathway)
• C5 is cleaved to produce C5b and from this
point on, both the alternative & classical
pathways are identical.
MANNAN – BINDING LECTIN
(MBL) PATHWAY
 Mannan – Binding Lectin (MBL)
Pathway

• Does not require antibody for activation

• Activation: REQUIRES MANNOSE –
BINDING OR MANNAN – BINDING
LECTIN (MBL)
– MBL is an acute phase protein
– It is produced in the liver, normally present in
the serum but increases during an initial
inflammatory response.
 Mannan – Binding Lectin (MBL)
Pathway
• It plays an important role as a defense
mechanism in infancy, during the interval
between the loss of maternal antibody and the
acquisition of a full – fledged antibody response.
 Mannan – Binding Lectin (MBL)
Pathway
• Lectins – proteins that bind to
carbohydrates

• Starts when MBL binds to mannose or

related sugars found glycoproteins or
carbohydrates of bacteria, yeasts, viruses
and some parasites in a calcium –
dependent manner.
 Mannan – Binding Lectin (MBL)
Pathway
• The structure of MBL is similar to that of C1s,
and it is associated with three MBL – serine
proteases (MASPs): MASP – 1, MASP – 2,
and MASP – 3.
– MASP – 1 and MASP – 3 have unclear functions
at this time.
– MASP – 2 is homologous to C1s and thus it
cleaves C4 and C2.
• Once C4 and C2 are cleaved, the rest of the
pathway is identical to the classical pathway.
 Complement Components and their
Functions
CLASSICAL PATHWAY
C1q Binds to Fc of IgM and IgG
C1r Activates C1s
C1s Cleaves C4 and C2
C4 Part of C3 convertase (C4b)
C2 Binds to C4b to form C3 convertase
C3 Key intermediate in all pathways
C5 Initiates membrane attack complex
C6 Binds to C5b in MAC
C7 Binds to C5bC6 in MAC
C8 Starts pore formation on membrane
C9 Polymerizes to cause cell lysis
Complement Components and their
Functions

Alternative Pathway
Factor B Binds C3b to form C3 convertase
Factor D Cleaves factor B
Properdin (Factor P) Stabilizes C3bBb (C3 convertase)
 Complement Components and their
Functions

MBL Pathway

MBL (serum) Binds to mannose (or other related

sugar)

MASP – 1 and MASP – 3 Unknown function

MASP – 2 Cleaves C4 and C2 (homologous to

C1s)
REGULATORS OF
COMPLEMENT ACTIVITY
 Classical Pathway & Lectin
Pathway
• C1 inhibitor (C1INH)
– inactivates C1 by binding to the active sites of
C1r and C1s (C1r and C1s become instantly
and irreversibly dissociated from C1q); also
inactivates MASP – 2
• Soluble C4b binding protein (C4BP) act in concert with
• complement receptor type 1 (CR1) factor I to inhibit C3
convertase
• membrane cofactor protein (MCP) formation.
• decay accelerating factor (DAF)
 Alternative Pathway
• Factor H – cofactor with factor I to
inactivate C3b and prevents binding of
Factor B to C3b

• Factor I – cleaves C3b and C4b

COMPLEMENT DEFICIENCIES
• C3 deficiency – most serious deficiency
• C2 deficiency – most COMMON
deficiency
 COMPLEMENT DEFICIENCIES

DEFICIENT COMPONENT ASSOCIATED DISEASE

C1 (q, r or s) SLE – like syndrome; recurrent infections
C2 SLE – like syndrome; recurrent infections,
atherosclerosis
C3 Severe recurrent infections;
GLOMERULONEPHRITIS
C4 SLE – like syndrome

C5 – C8 Neisseria infections

C9 No known disease associated

C1INH HEREDITARY ANGIOEDEMA

 COMPLEMENT DEFICIENCIES

DEFICIENT COMPONENT ASSOCIATED DISEASE

Factor H or factor I Recurrent pyogenic infections
MBL Pneumococcal diseases, sepsis, Neisseria
infections
Properdin Neisseria infections
MASP – 2 Pneumococcal diseases
DAF Paroxysmal nocturnal hemoglobinuria
MIRL Paroxysmal nocturnal hemoglobinuria
 Nonimmunologic Classical
Pathway Activators
• Bacteria (E. coli and Salmonella)
• Viruses (parainfluenza virus, HIV)
• Apoptotic cells
– Interact with C1q directly causing C1 activation
(non-ab and ag activation)
• Urate crystals
• Myelin basic protein
• Denatured DNA
• Bacterial endotoxin
• Polyanions