Complement

Kathleen Basa
Kris Nicole de Guzman
Jessica Mae Ong
3HMT
 Immune System

Innate Immune Adaptive

System Immune System
Complement
Heat labile series of enzymes or proteinases

Circulating inactive precursors or pro-proteins

Kills invading microorganism directly & supplements immune response

Malfunction may cause injury to cells, tissues, & organs

 Provides
Provides
actual
actual
protection
protection
from the
from the
response
response

The Antibodies
Antibodies
point out

Complement
point
the out
target,
the target,
complemen
complemen
t destroy it
t destroy it

System
C1, INH,
Bacterial
Inactive
Factor 1, H,
C1, INH,
Bacterial
invasion
Complement
Inactive
c4-bp
Factor 1, H,
invasion
Complement
c4-bp

A
ct
iv
at
io
n
o
f
C
o
m
pl
e
m
e
e
n
t
 complement

antibody

BACTERIAL SURFACE
antigen
Activation of Complement Pathways
• Classic Pathway • Alternative Pathway
C1 complex
C1q

C1r C1s
antibody

BACTERIAL SURFACE
antigen

• Mannose-binding lectin Pathway

PROTEINS
C1q
 Small letter after the number indicates that the protein is
smaller protein resulting from cleavage of a larger precursor by
a protease.

C3a and C3b

 Lowercase suffixes are fragments.
 Larger fragment is designated “b” and the smaller fragment
“a”, with an exception to C2a and C2b.
C4b
PROTEINS OF THE COMPLEMENT
SYSTEM
 C1
 C1 exists in blood serum as a molecular

complex containing:
 6 molecules of C1q
 2 molecules of C1r
 2 molecules of C1s
 The constant regions of mu chains (IgM) and

some gamma chains (IgG) contain a binding

site for C1q.
 Calcium-dependent complex
 C1q

C1r C1s
antibody

BACTERIAL SURFACE
antigen
C3
 C3 is the most abundant protein of the

complement system (~1.3 mg/ml)

 ability to activate itself

C5
 unique to the terminal complement pathway
 present in plasma at 70 mg/L.
 Together with the other proteins of the

terminal pathway, C5b forms the MAC.

Classical Pathway
Serum Protein MW, kD Concentration Function
ug/ml
C1q 410 150 Binds to fc region of
IgM and IgG

C1r 85 50 Activates C1s

C1s 85 50 Cleaves C4 and C2

C4 205 300-600 Part of C3 convertase

C2 102 25 Binds to C4b-forms

C3 convertase

C3 190 1200 Key intermediate in

all pathway

C5 190 80 Initiates membrane

attack complex

C6 110 60 Binds to C5b in MAC

C7 100 55 Binds to C5bC6 in

MAC

C8 150 55 Starts pore formation

 Properdin
 Constituent of normal serum with a

concentration of approx. 5-15 ug/ml

 Stabilizes the C3 convertase

 Factor D
 Plasma protein that circulates in active

enzyme form
 Cleaves factor B
Alternative Pathway

Serum Protein MW, kD Concentration Function

ug/ml
Factor B 93 200 Binds to C3b to form
C3 convertase

Factor D 24 1-2 Cleaves Factor B

Properdin 53 25 Stabilize C3bBb-C3

convertase
MBL
 MBL
 Binds to mannose or related sugars to initiate

the pathway
 An acute phase protein due to its presence in

the serum but increases during initial

inflammatory response
 Appearance of bouquet of tulips
 MASP-1 and MASP-2
 Activated when MBL bounds to the cell

surface
 95% of total MASP is unbound
 Homologous to C1r and C1s
MBL Pathway
Serum Protein MW, kD Concentration Function
ug/ml
MBL 200 1 (0.01-20) Binds to mannose

MASP-1 100 1.5-12 Helps to cleave C4

and C2

MASP-2 76 ND Cleaves C4 and C2

Fluid Phase Regulators
C1 inhibitor or C1INH
 Molecular weight of 150,000

 Inactivate C1 by binding to the active sites of C1r

and C1s
 Inhibits clot formation essentials

Factor H
 MW of 160,000

 Prevents binding of Factor B by binding to C3b

 Allows Factor 1 to break down C3B

Factor I
 A serine protease that inactivates C3b and

C4b when they are associated with Factor H

C4-binding protein or C4BP

 Cofactor for Factor 1 in the inactivation of

C4b
 Abundant in plasma
 causing cessation of the classical pathway
S protein
 Control protein that acts at a further level

complement activation
 Also known as VITRONECTIN
Cell Bound Regulators
 Binding to regulators

 Found on host-cell membranes to protect the

cell from bystander lysis
CR1 or CD35
Found mainly in the peripheral blood

Greatest affinity for C3b

Function as cofactor similar to Factor H

Act as a receptor on platelets and RBC

Mediates
Mediates transport
transport of
of C3b-coated
C3b-coated immune
immune complexes
complexes to
to the
the liver
liver
CD55 or DAF
Wide tissue distribution

Dissociates
Dissociates both
both classical
classical and
and alternative
alternative pathway
pathway C3
C3 convertases
convertases

Protect host cell from bystander lysis

Classify self from nonself

 C3b Bb

DAF
CD59 or MIRL
Block the formation of MAC

Bind C8 and prevent insertion of C9 into the cell

CD46 or MCP
Carried by platelets, monocytes, B & T lymphocytes

Most
Most efficient
efficient cofactor
cofactor for
for Factor
Factor II mediated
mediated cleavage
cleavage of
of C3b
C3b

Inhibit
Inhibit C3b,
C3b, thus
thus limiting
limiting the
the conversion
conversion of
of enzyme
enzyme C3
C3 converatse
converatse
Pathways of
Complement
Activation
A. Classical Pathway

 Complement components involved: C1, C2, C3,

C4, C5, C6, C7, C8, C9
 Stimulus: Antigen-antibody complex
 Immunologic Activators: IgM or IgG (3, 1, 2)
 Non – immunologic Activators: Apoptotic cells,

Staphylococcal protein A, C-reactive protein,

Gram negative bacteria, certain viruses and
parasites
●
Needs two molecules to
activate the pathway
●
Binds to CH2

●
One molecule to activate the
pathway
●
Binds to CH3
●
Most efficient
3 STAGES of
the Classical
Pathway
1. Recognition Unit

C1
-first complement
component to bind
- 750, 000 D
◦C1 complex: C1q, C1r
C1s -> stabilized by
calcium
 - one subunit

C1q
- 410, 000 D
- six strands that form six
globular heads with a
collagenlike tail portion
- “recognizes” the Fc region
of the two adjacent
antibody molecules
- binds to the antibody
- at least two of the
globular heads must be
bound to initiate the
pathway
C1R & C1S
- two subunits each
(Clr, Cls)2 complex – S-shaped
structure
C1s – C1r – C1r – C1s
- generate enzyme activity
to begin the pathway
- serine protease
proenzymes
 Binding of C1q 
activation of C1r & C1s
◦ Activated C1r’s substrate:
C1s
◦ C1s’s substrate: C4 & C2
 2. Activation Unit

C1s cleaves C4 to split

off C4a

Splitting off of C4a

 2. Activation unit

C2 is cleaved by C1s

C2b is released
 2. Activation Unit

C3 cleaves to the C4bC2a

complex

C3 is cleaved into C3b &

C3a
2. Activation Unit
 3. The Membrane Attack Complex

C5a splits off

C5b attaches to the cell

membrane
 3. The Membrane Attack Complex

C8 binds and forms a small

hole in the membrane

C9 binds to the C5-C8

complex
B. Alternative Pathway

 “Incomplete Pathway” or “Properdin pathway”

 Immunologic Activators: Aggregated IgA, in

some instances IgG4 and IgE

 Non Immunologic Activators: Bacterial and

plant polysaccharide, LPS, zymosan, Inulin,

Cobra-Venum Factor, viruses and tumor cells
 B. Alternative Pathway

C3 in plasma and cleaved

C3b is cleaved and

becomes stable
 B. Alternative Pathway

Factor B binds to C3b

Factor D binds to the

complex and Ba is lost
B. Alternative Pathway

Properdin stabilizes C3
convertase

C3b is formed
continuously
 B. Alternative Pathway

C5 binds to C5
convertase

C5 splits into C5a & C5b

C. Mannose – Binding Lectin pathway

 Mannose-binding lectin – belongs to the family of

calcium-dependent lectins, the collectins
◦ Homologous in structure to C1q
◦ A lectin that binds mannose or related sugars to
initiate the pathway
◦ Normally present in serum
◦ Increases during an initial inflammatory response
 C. Mannose – Binding Lectin pathway
MBL
MASP-1
Two binds& to
MBL-associated
serine proteases
the
MASP-2surface
(MASP-1 cleave
& MASP-2)
C4
of a& cell
become C2
activated

C
3
c
o
n
v
er
ta
s
e
is
fo
r
m
e
d
 Consequenc
es of
Complement
Activation

Chem
otaxis

Inflamma
tory
Cell
Reaction lysis

Opsoniz
ation
Alterations in Complement Level
 May occur:
 due to persistent infection
 Autoantibody response to self antigens

 May contribute to the pathology of the

disease
 Activation is related intravascular

thrombosis, which leads to ischemic injury to

tissues
Elevated Complement Levels
 Inflammatory conditions
 Trauma
 Acute illness
 Common and nonspecific
Decrease Complement Levels
 Low levels due to:
 Excessive activation
 Being consumed
 Genetic defect

Hypocomplementemia
- Due to the complexing of IgG or IgM
antibodies capable of activating complement
 Increase susceptibility to pyogenic Infections:
1. Deficiency of the opsonic activities of
complement
2. Any deficiency that compromises the lytic
activity of complement
3. Deficient function of the mannose-binding
lectin pathway
Measurement of Active
Components of the
Complement
 Complement components
can be assessed by
nephelometry.

 Nephelometer is a device
that measure the size or
density of solid particles
present in a liquid.
 C1 Esterase Inhibitor
 measures the activity and concentration of C1
inhibitor in serum

 Deficiency: Hereditary angioedema (HAE)

Lupus nephritis
 C1r, C1s, C2-C8
 Homozygous deficiencies

 Autoimmune disease (like SLE), arthritis,

chronic glomerulonephritis, infections, and
vasculitis
 C1q
 Evaluated in serum
 Decreased level:

◦ hypocomplementemic urticarial vasculitis

◦ severe combined immunodeficiency or X-linked
hypogammaglobulinemia
Associated Disease:
◦ SLE-like syndrome
 C1q Binding

 Measures binding of immune complexes

containing IgG1, IgG2, or IgG3 and IgM to
C1q

 Useful as prognostic tool at diagnosis and

remission of acute myelogenous leukemia
 C2
 Most common complement deficiency
 Autosomal recessive disorder (C2 gene is on

chromosome 6 in MHC)
◦ 50% of patients have no symptoms
◦ 50% have an infection of S. pnemoniae, N.
meningitidis and H. influenzae and exhibits lupus-
like disorder with photosensitivity and rash.
 C3
 acute-phase protein
 Elevated level: acute inflammatory disease

 Extremely decreased level: patients with

poststreptococcal glomerulonephritis
and with inherited C3 deficiency.
 Decreased level: severe liver disease, SLE

with renal disorder

 C3b Inhibitor
 Low C3 levels
 Absence of C3PA in serum
 High C3b levels
 Deficiency causes an increase susceptibility to

infection
 C3PA (Properdin Factor B)
 Decreased level of C3 results from classic or
alternate pathways of complement activation.
◦ Decreased levels of C3 and C4, activation of classic
pathway
◦ Decreased levels of C3 and C3PA with normal C4
indicates activation via alternate pathway
◦ Activation of Classic pathway
 immune complex diseases
 various forms of vasculitis
 acute glomerulonephritis.
◦ Activation of Alternate pathway
 Chronic hypocomplementemic glomerulonephritis
 Disseminated intravascular coagulation (DIC)
 Septicemia
 Subacute bacterial endocarditis
 PNH
 Sickle cell anemia

•Activation of both pathways

• Systemic Lupus Erythematosus
 C4
 most sensitive indicator of disease activity
 an acute-phase reactant
 Increased level: acute inflammatory reaction
 Decreased level with high anti-n-DNA and

antinuclear antibody titers, confirms SLE

 Classic pathway activation,C4 destroyed.
 C4
 Extremely low C4 with normal C3 indicates
genetic deficiency of either C1 inhibitor or C4
 Reduction of C3 and C4 implies activation of

classic pathway
 C5
 Causes an increased susceptibility to bacterial
infection
 Autoimmune disorder (e.g. SLE)
 Leiner’s disease
 C6
 Decrease C6 makes individual susceptible to
significant Neisseria infections.
 Associated Disease:

◦ SLE
◦ Raynaud’s phenomenon
◦ Scleroderma-like syndrome
◦ Vasculitis
 C7
 Decreased level:
◦ Raynaud’s phenomenon
◦ sclerodactyly
◦ telangiectasia
◦ severe bacterial infections caused by Neisseria
species.
 C8
 Decreased C8 is associated with SLE and
makes patients highly susceptible to
Neisseria infections.
 Associated Diseases:

◦ Neisseria infections
◦ Xeroderma pigmentosa
◦ SLE-like syndrome