Kim Alyssa B.

Go UIC MLS-3E
INNATE IMMUNITY III: COMPLEMENT
COMPLEMENT ACTIVATION
IMMUNOLOGY-SEROLOGY LEC.
● Involves three separate pathways:
DR. ALFREDO HINAY
○ Classical Pathway
○ Alternative Pathway a.k.a. Properdin
GENERAL CHARACTERISTICS OF COMPLEMENT Pathway
● Heat labile (heat sensitive) series of more than ○ Mannose-binding Lectin (MBL) pathway
30 soluble plasma proteins
○ Some are enzymes or proteinases
○ Major fraction of beta globulins is
complement
● Part of the humoral component of the Innate
Immune System
○ Participates in the INFLAMMATORY
RESPONSE
● Interacts in a specific way to enhance host
defense
● First described by Paul Ehrlichs
● Major role:
○ Promotes OPSONIZATION and LYSIS
of foreign substances
CLASSICAL PATHWAY
● The recognition unit
NOMENCLATURE ○ C1 molecule (composed of C1q, C1r,
● Activated C1s cleaves C4 to C4a and C4b, and C1s)
which binds to the microbial surface ○ Interlocking enzyme system
● Named with a capital C and a number: ○ Stabilized by calcium ions
○ Ex. C1, C2 ■ This is the difference between
● Small letter after the number: antibodies because antibodies
○ Ex. C4b, C4a are not calcium-dependent
■ Indicates that the protein is ● Requirement for Activation:
smaller portion of a large ○ Antigen and Antibody Interaction
precursor as a result of ○ C’ binds to CH2 domain
cleavage by proteases ○ All IgM can fix complement
■ Larger fragment: indicate by ■ According to Stevens, the
small letter “b” immunoglobulin that is most
■ Smaller fragment: indicated by potent in activating the
small letter “a” Classical Pathway is the IgM
(because it is pentameric)
9 MAJOR COMPLEMENT COMPONENTS INVOLVED ■ Pentameric; therefore, it has a
IN THE ACTIVATION: lot of FAB regions that are
● Most are produced in the LIVER Except: exposed that will bind to your
○ C1: produce by intestinal epithelial antigen
cells ○ All IgG can fix complement except IgG4
○ Factor D: produced by adipose tissues ■ According to Stevens, the
● Macrophages and Monocytes: additional BEST IgG that can activate the
sources of C1, C2, C3 and C4 Classical Pathway is the IgG 3
● The rest of the components ● Followed by IgG 1
○ Acts as stabilizers and control proteins, ● Then IgG 2
and proteases ■ Can be used in the absence of
IgM (in the classical Pathway)
 Kim Alyssa B. Go UIC MLS-3E
■ Needs at least 2 monomers of ■ C2a
IgG attached to target cells ● Larger portion
■ The distance between Ab1 and ● Binds to the other large
Ab2 should be around 30-40 portion of the cleaved
nm. (C1q) C4, C4b to form an
● The reason why we enzyme called C3
should secure the convertase (C4b2a)
distance is because ● C4 - THE SECOND MOST ABUNDANT
C1q, specifically the PROTEIN/ SOLUBLE COMPONENT OF THE
head portion of q will COMPLEMENT.
bind to the antibody. ○ C4 is also the specific substrate for the
● How can we secure C1qrs
that the distance ○ C3 is the 1st one
between the two ● C3 convertase (C4b2a)
antibodies is 30-40 ○ An enzyme that will try to convert
nm? (cleave) C3 to C3b and C3a in a higher
○ To know this, loop.
we need to ■ This means that the conversion
secure that the of C3 is faster
produced ■ It is important to convert many
antibodies (IgG) C3b because it is a potent
should be at opsonin.
least a ● C3b will also attach to
minimum of other parts of the
1000 IgG that bacterial structure.
will bind to the ○ C3a
antibody ■ Smaller portion
● Only C1s has the enzymatic portion that will ■ Becomes the anaphylatoxin
cleave first C4, followed by C2 ○ C3b
● C1qrs- (cleaves) only has 2 substrates: ■ Larger portion
○ C1s is the one who does the cleaving ■ Will bind to the C4b2a to
○ C4 become C4b2a3b (C5
■ Most abundant among the two convertase)QQQ
so, it is cleaved first ● C5 convertase (C4b2a3b)
■ Once cleaved, C4a becomes ○ An enzyme that will try to convert
Anaphylatoxin. C4b will bind to (cleave) C5
the cell surface of the bacteria. ○ C5a
■ Anaphylatoxin : The smaller ■ Smaller portion
portion of the cleaved ■ Becomes the anaphylatoxin
complement (C2b & C4a) ○ C5b
○ C2 ■ Larger portion
■ It is very unique; among all the ■ Will try to recruit C6 and C7
complement components, C2, ● The recruitment of C6
once cleaved, C2b is smaller in and C7 by the C5b is
proportion compared with the the first phase of
C2a. membrane attack
■ C2b complex (C5bC6C7
● Smaller than the C2a complex)
● Will serve as the ● C8 will then be recruited
anaphylatoxin that will and will penetrate the
recruit other immune cell membrane
cells.
 Kim Alyssa B. Go UIC MLS-3E
○ C8 will try to ○ C3a becomes anaphylatoxin while C3b
penetrate the becomes free floating
cell membrane ● The free floating C3b will bind to Factor B.
which will be ○ This binding will lead to the activation of
polymerized by Factor D (unstable)
the C9 ■ Since Factor D is unstable, it is
■ After stabilized by Magnesium.
this, C8 ○ Factor D
will be ■ Unstable
remove ■ Stabilized by Magnesium
d, ■ Function: To cleave Factor B
forming ● Note: Factor B will only
(bacteri be cleaved when it is
a)cell bound to C3b. It is not
lysis. cleaved when free
floating
■ Factor B is cleaved into two
portions: Ba and Bb
○ Factor B
■ Ba
● Will activate Factor D to
cleave another C3bB
complex
● Will also serve as
anaphylatoxin
■ Bb
● Combines with C3b to
form C3bBb complex
○ C3 convertase (C3bBb)
■ It is Unstable
■ To become stable, it should be
added with Properdin
● This is why the
Alternative Pathway is
The Classical pathway also called the
Properdin Pathway
■ After being stabilized, it will
cleave C3 on a high loop
THE ALTERNATIVE PATHWAY ● C3a becomes
● Have almost the same activation with the anaphylatoxin
classical pathway. ● C3b will bind to the
○ The difference is only on the recognition C3bBbP forming
units C3bBbPC3b complex
● Divide into 3 parts: (C5 Convertase)
○ Recognition Unit ○ C5 Convertase (C3bBbPC3b)
○ Activating Units ■ Will cleave C5
○ MAC (membrane attack complex) ● C5a: anaphylatoxin
● According to Stevens, In the Alternative ■ C5b: recruits C6 and C7
pathway, the C3 is spontaneously hydrolyzed forming C5bC6C7 complex
○ This means that naturally, even without ● Allows C8 to penetrate
infection, C3 is cleaved. on the cell membrane
○ Forms C3a and C3b
 Kim Alyssa B. Go UIC MLS-3E
● C8 will then recruit the
polymerization of C9
which will lead to the
removal of C8 causing a
bacterial cell lysis. Then COMPLEMENT REGULATORY PROTEINS (CRP)
MAC is formed. ● C1 Inhibitors (C1 INH)
○ Controls the activity of C1
MBL PATHWAY (Mannose Binding Lectin) ○ Binds to C1 and can dislodge bound C1
● Initiated by microorganisms with mannose or to immunoglobulins
similar sugars in the cell wall or outer ■ C1 activation is halted
membrane. ● When there is no
● Almost the same with classical pathways interaction with C1q to
specifically on the recognition units. CH2 part of antibody
○ CP: C1qrs structure, C1r and C1s
○ MBLP: MASP-1, MASP-2, MASP-3 will not be activated
■ MBL is homologous to CP’s C1q ● Factor I - Inhibitor of C4b
● Binds to the invading ○ Cleaves C4b into C4c and C4d
cell ● C4 binding protein (C4BP)
● MASP-1 is not initiated with antibodies ○ Destroys bound C4b and can also
○ Even without antibodies, MBL Pathway displace C2a from the classical
can still be activated pathway C3 convertase (C4b2a)
● MASP-2 is homologous with CP’s C1s ● Inhibitors of C3 Convertase formation
○ Among MASP 1,2,3, MASP-2 has an ○ Factor I inactivates C4b and C4b
enzymatic action against C4 and C2 ■ Converts C3b to iC3b(inactive)
● After this, everything is the same with the ● When C3b is inactive, it
classical pathway. can no longer bind to
C4b2a complex and it
1. Lectin Binds to an invading cell will not form C5
2. Bound lectin splits C2 into C2b and C2a and C4 convertase
into C4b and C4a ■ Requires Factor H as cofactor
3. C2a and C4b combine and activate C3 ○ Factor H and C4bp dissociates C3
convertases of both classical and
alternative pathways
● Regulation of MAC (membrane attack
complex)
○ S-protein (vitronectin)
■ Binds the C5b-7 complex,
thereby preventing its insertion
in the cell membrane
● When there is no
C3- CENTRAL MOLECULE OF C’ ACTIVATION C5bC6C7 complex, C8
PATHWAYS will not be recruited
● C3 is the major constituent of the complement ■ Forms sC5b-9 complex
system and is present in the plasma at a ● Advantage: no cell lysis
concentration of 1200 ug/mL ○ Clusterin - a.k.a Sp-40 or apolipoprotein
● Pivotal point for all three pathways J
○ Cleavage of C3 to C3b represents the ■ Prevents insertion of the C5b-7
most significant step in the entire complex into the cell membrane
process of complement activation. (Similar in action of S-Protein)
 Kim Alyssa B. Go UIC MLS-3E
DISEASES ASSOCIATED WITH DEFICIENT C’
COMPONENTS (MEMORIZE!!!)

Deficient Component Associated Disease

C1 SLE-like syndrome

C2 SLE-like syndrome,
recurrent infections

C3 Severe recurrent
infections

C4 SLE-like syndrome

C5-C8 Neisseria syndrome

C9 No known disease
association

C1 INH Hereditary angioedema

Factor H Recurrent bacterial

DESTRUCTIONS OF COMPLEMENT IN VITRO