Complement

http://www.umich.edu/~projbnb/imm/complement.swf

Contents
Part Indroduction Part Activation of complement system Part Biological functions of complement Part Regulation of complement system

Section I introduction
1 Discovery of complement 2 Definition of complement system

3 Composition of complement system

4 name of complement system

Discovery of complement

The end of 19 century Jules Bordent (1870-1961),

bacterium

Fresh Antiserum

Lysis

Conclusion Fresh antiserum can agglutinate and lyse bacterium

bacterium

Normal serum

heated to 56 C, 30min Agglutination but no lysis lysis

Conclusion The heated antiserum can agglutinate bacterium but cannot lyse bacterium

The bacteriolytic activity requires two different substances

Which can agglutinate bacterium Which can lyse bacterium

Agglutinin, now as antibody

Complement completing the action of antibody

Heat-labile Innate proteins in serum

Section I introduction
1 Discovery of complement 2 Definition of complement system

3 Composition of complement system

4 name of complement system

What is it?
A series of proteinase in serum or on cell menbrane.

What is the fuction of it

To lyse bacteria and cells through membrane attack complex. As a bridge both the innate and adaptive immunity systems

Difinition of complement system

A system of serum and cell surface proteinase including more than 30 proteins ) that interact with one another to mediate cell lysis and with other molecules of the immune system to generate important function of innate and adaptive immune response .

Section I introduction
1 Discovery of complement 2 Definition of complement system

3 Composition of complement system

4 name of complement system

1. Intrinsic components of complement

Classical pathway: C1 (C1q,C1r,C1s),C2C4 MBLmannan-binding lectin) pathway: MBL, MASP( MBL-associated serine proteinase) Alternative pathway: B factor, D factor, properdin Common terminal pathwayC3C5C6-9

2. complement regulatory proteins

C1 inhibitor, I factor, C4-binding protein, H factor,etc

3. Complement receptors
CR1-CR5

Section I introduction
1 Discovery of complement 2 Definition of complement system

3 Composition of complement system

4 name of complement system

Intrinsic components in classical pathway:C1~C9 Intrinsic components in alternative pathway: factor D Regulatory proteins:C1 inhibitor (C1INH) Cleaved fragments:C3a, C3b; Activated components:C1,C4b2a

Contents
Part Indroduction Part Activation of complement system Part Biological functions of complement Part Regulation of complement system

Classic pathway
Alternative pathway

Three pathways
MBL pathway

The activation of complement

The soluble proteins of the complement system are secreted as inactive forms called zymogens.

The cleavage of a zymogen usually produces a large active fragment with enzymatic activity and a small fragment with inflammatory effects.

Cascade reaction the sequence of reactions, each being the catalyst for the next, that

leads to the terminal complement pathway and cell lysis

Summary of Complement Pathways

classic
Antibody-antigen complex, binds C1

MBL
mannose binding protein, binds MASP1

alternative
most primitive, autoactivation of C3

Formation of C3 convertase and C5 convertase Common terminal pathway (Formation of membrane-attack complex )

Classical pathway

1.Initiating substances: antigen-antibody complex or immune complex 2.Components: C1,C4,C2,C3,5,6,7,8,9

C1q C1r C1s + C1

C1q + 2 C1r+2C1s = C1

CH2
Ag

No activation

activation

The C1q must bind at least 2 CH2 or CH3 domains

staple form

because IgM is a pentamer, a single IgM can activate C1q IgM is the best complement activator

IgG is monomer,So about 1000 molecules of IgG are required to assure that two IgG are close enough to each other to initiate C1q binding.

3 Process
1). initiation phase

activate C1 molecule

IgG1-3 and IgM can activate complement by classical pathway C1q binds to the Fc part of the antibodies, CH2 of IgG or CH3 of IgM, and then activate C1r which cleave C1s

Bacteria with antigenic proteins on the surface

After antibody binds to antigen on the surface of a pathogencomplement-binding sites are exposed

globular domains of C1q binds to at least two IgG molecules ,which induce a conformational change in C1r.

C1r convert to an active enzyme. Then,C1r cleaves C1s to an active enzyme

2. amplification phage Formation of C3 convertaseC4b2a) and C5 convertase(C4b2aC3b)

The activated C1s cleaves C4 into C4b and C4a.

The C4b fragment binds to the surface of the pathogen.

C4b

C4a

C1s then cleaves C2 into C2b and C2a.

C2

C4b

C2b
C2a

C4b

C4b and C2a together form the C3 convertase(C4bC2a)

C2a

C4b C3 convertase

C3 convertase cleaves C3 to C3a and C3b.

One C3 convertase can produce many C3b. C3b can bind to C4b2a to form C4b2a3b

C3a

C3 convertase

C4b2a3b complex,C5 convertase

C5 convertase

3). Effector Phase (Common terminal pathway)

Formation of the Membrane Attack Complex (MAC)

MAC: C56789

C5 convertase cleaves C5 into C5a and C5b.

C5a

C5b

C5 convertase

C5b binds to the surface of target cell and provides a binding sits for the components of MAC C5b is labile unless C6 bind to it

C6

C5b

Then C7 binds. C7 inserts into the phospholipid

bilayer of the membrane.

C7

C6 C5b

Then C8 binds to the complex and also inserts into the bilayer,creates a small pore.

C6

C7 C5b

C8

Finally, C9 molecules bind to the complex and polymerize.

C9

C6

C7 C5b C8

Ten to sixteen C9 molecules form a large pore in the membrane

C6

C7 C5b C8

C6

C7 C5b C8

C2 Ab+Ag C1

C4,

C4b2a (C3convertase)

C3
C4b2a3b

(C5convertase)

C5 MAC C5b,C6,C7,C8,C9

Alternative pathway

1.The initiating substances: some components of microbial cell surface aggregated IgA or IgG4 ---providing a protective surface for binding of complement 2.Components: factor D, factor B, properdin,C3,5,6,7,8,9 3.Function: join in nonspecific immunity

 Normally, C3 in plasma is being continuously cleaved at a low rate to generate C3b in a process A small amount of the C3b in the fluid phase is unstable and inactive C3b may attach to the surfaces of cells, including normal cells or microbes

C3b attaching to normal cells will be hydrolyzed by I factor and membrane cofactor protein which cannot be expressed on microbes. C3b attaching to microbes may remains active for a longer time.

The process of complement activation in alternative pathway:

1. Initiation step: C3b binds to microbe surface,then binds factor B, and forms C3 convertase. 2. Activation step: form C5 convertase 3. Effector step: membrane-attack complex (MAC) , C5-9

Lectin or MBL Pathway

Mannose-binding lectin (MBL)

MBL:an accute phase protein in inflammation

1.Initiating substances: MBL combine with mannose on the surface of microbe. 2.Components: Mannose-binding lectin (MBL) , MBL-associated serine protease (MASP) C4,C2,C3,5,6,7,8,9 3. Function: join in nonspecific immunity

Mannose-binding lectin C1q-like

MBL is Structurally similar to C1q

The MBL and c-reactive protein were produced by liver cells which we call as stress protein after microbe infection

MBL binds to mannose on the surface of microbes. Then MASPs bind to it.

MASP-2 MASP-1

MASP-2 Similar to C1r,C1s MASP-1

glycoproteins

Bacterium

C4b
C2a

C4a
C2b

glycoproteins

Bacterium

SUMMARY OF COMPLEMENT ACTIVATION

Classical Pathway

Lectin-Binding Pathway

Alternative Pathway

C1q

MASP

C3

[C4b2b]

[C3bBbP]

C3 Convertase

C3a

C3b

C5a

C5b

C5b-C 9 (membrane attack complex)

Cell Injury

Contents
Part Indroduction Part Activation of complement system Part Biological functions of complement Part Regulation of complement system Part Complement and disease

Biological functions of Complement

1. Lyse bacteria and cells
Classical Pathway Lectin-Binding Pathway Alternative Pathway

2. Opsonization
3. Immune adherence (Elimination of immune complex)

C1q

MBP

C3

[C4b2a]

[C3bBbP]

C3 Convertase

4. Induce inflammation reaction

C3a Anaphylatoxins Chemokine C3b C3b (opsonlzation Immune adherence)

C5a

C5b

C5b-C 9 (membrane attack complex) Cell Injury

1. Lyse cells, bacteria and virus The complement system lyses target cells bacteria,virus and other microbes through alternative pathway and MBL pathway of complement-activating in absence of antibody and through classical pathway in presence of antibodies

MAC PORES

Source: undetermined

Source: undetermined

Source: www.wikimedia.org

2. Opsonization
The phenomenon that phagocytosis of phagocyte are enhanced by antibody or complement binding to microbial surface. The bound C3b ,C4b to CR1 on macrophage and neutrophils enhances their actions

Opsonization: C3b,C4b ----CR1

3. Immune adherence -----Clearance of immune complex

C3b,C4b----CR1
C3b or C4b coats immune complexes helping binding to CR1 on erythrocytes, which carry the immune complexes to the liver or spleen where they can be removed and phagocytized

4. Mediate inflammatory reaction:

Anaphylatoxins : C3a, C4a,C5a

Chemokine : C3a, C5a

Anaphylatoxins
mast leukocyte

C3a

C5a
degranulation
Containing Histamine,
leukotriene,etc

allergy

C5a

Contents

Part Indroduction Part Activation of complement system Part Biological functions of complement

Part Regulation of complement system

The explosive potential of the complement system requires that it be kept under tight control.

Mechanism of complement regulation

I. regulation of self-inactive :decay

II. action of regulatory factors

I. regulation of self-inactive : decay (hydrolysis)

C3 convertase: C4b2a or C3bBb

C5 convertase : C4b2a3b or C3bnBb C5b,C3b,C4b

negative feedback

II. action of regulatory factors

Inhibit activation: classical pathway

C1 inhibitor (C1INA): plasma protein Plasma proteins: Factor I Cell membrane proteins: - decay accelerating factor (DAF): - membrane co-factor protein (MCP):

inhibit C3 convertase:

Inactivate anaphylatoxins: cleave C3a and C5a serum carboxypeptidase N (SCPN): Inhibit MAC: Protectin (CD59): cell associated protein

SUMMARY OF COMPLEMENT ACTIVATION

Classical Pathway

Lectin-Binding Pathway

Alternative Pathway

C1q

MBP

C3

C1INA

[C4b2b] [C3bBbP]

Hydrolysis DAF-cell

C3 Convertase

C3a

C3b

Factor I MCP-cell

SCPN C5a C5b

C5b-C 9 (membrane attack complex)

Protectin-cell

Cell Injury

C1 inhibitor

C2

DAF -----binds to C4b and C2a, preventing formation of C3 convertase

C4b

C2a

MCP (Membrane
cofactor protein)

C4c

I factor

C4d
C4

I factor and MCP have the same fuction on C3b

Protectin(CD59)

MAC
Protctin
C6

C7 C 9 CC 9 9C 9

MCP, DAF, Protectin

Can be expressed on normal cells, not on microbes

Thus
On normal cells, complement system can not form MAC