Biomaterials Research - Manfred Maitz

The Complement System

and Kinin System
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

Background
Evolutionary old system
Non specific defence system
Constitutive
Very fast
Selectivity and self-tolerance
Activation by antibodies possible
Interaction with the specific immune system
Functions
Lysis of cells/ bacteria and viruses
Opsonization for phagocytes
Immune Clearance
Similarities and crosstalks with the clotting cascade
Cascade of serine proteases, which activate each other
Two (three) pathways of activation
Factor XIIa (Hageman-Factor) activates both the clotting cascade
and the complement cascade
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
Biomaterials Research - Manfred Maitz

The Membrane Attack Complex

C5a

C5

70-100

C6 C5b C7
C9
C9
C8 C9 C9

C9
C9C9

C9C9
C9
C9
 Biomaterials Research - Manfred Maitz

The Complement Cascade

The Alternative Pathway C3 P C3a

Ba Properidin
B
C3b C3bB C3bBb C3bBbP C3bBb3b
C5a
D

C3a B

C3iBb C3iB C3i C3 C5

Ba
The Tick-over Activation
C3

The Classical Pathway

C5b
Antibody +
C1qrs

C4 C4b C4b2a C4b2a3b

C2 C2b C3 C3a
C4a
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

Regulation of the Complement Cascade

Short half-time of
C3b
C3bBb
C5b
C1 inhibitor
Inhibits the C1s activity
Protein S in Serum
Binds to C5b67
Inhibits Formation of the Membrane Attack Complex
HRF or CD59
Bind to C8
Inhibits C9 binding
Factor H
Binds to C3b
Facilitates binding of Factor I
cleaves C3b to inactive iC3b
cleaves C4b to inactive fragments
Decay Accelerating Factor
Increased dissociation of C3 convertase (both pathways)
 Biomaterials Research - Manfred Maitz

Complement Activation
General
Hydrophobic surfaces
Oxides
Strong binding of C3(b) to nucleophilic groups (-NH2, -OH)
Higher absorption of C3 to crystalline TiO2 than to amorphous
Kallikrein directly activates C5
Plasmin directly activates C5
Classical Pathway
Antibodies IgM, IgG1, IgG2, IgG3
Lectin via the mannan binding protein (MBP) Lectin Pathway
Hageman Factor (F XIIa)
Rough surfaces
C-reactive protein (CRP)
(Zirkonium, transiently)
Alternative Pathway
PE debries
Acetylated chitosan
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

Complement Receptors
Receptor Ligand Cellular distribution
CR1 (CD35) C3b>iC3b B-Cells
C4b Phagocytes
RBC
follicular dendritic cells

CR2 (CD21) iC3b B cells

C3dg epithelial cells

CR3 (CD18/11b) iC3b Phagocytes

Zymosan NK Cells
ICAM-1 follicular dendritic cells

CR4 (CD18/11c) iC3b Phagocytes

 Biomaterials Research - Manfred Maitz

Anaphylatoxins
Fragments C5a, C3a, C4a
Degranulation of Phagocytes
Reactive oxygen species
Prostaglandins
Monocytes
IL-1, IL-6
Mast cells
Histamine
Chemotaxis
Only C5a
 Biomaterials Research - Manfred Maitz

Consequences in vitro
Lysis of innocent neighbour cells
Red blood cells
Activation of phagocytic cells
Release of reactive oxygen species
Release of mediators
 Biomaterials Research - Manfred Maitz

Consequences in vivo
Factors of complement activation at revised hip implants
One single study (Tang L. et al. J Biomed Mater Res 41: 333-340 (1998))

Au-Mercaptoglycerol induces strong inflammatory

response in control animals.
No reaction in Complement-depleted animals.
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

The Complement Cascade

The Alternative Pathway C3 P C3a

Ba Properidin
B
C3b C3bB C3bBb C3bBbP C3bBb3b
C5a
D

C3a B

C3iBb C3iB C3i C3 C5

Ba
The Tick-over Activation
C3

The Classical Pathway

C5b
Antibody +
C1qrs

C4 C4b C4b2a C4b2a3b

C2 C2b C3 C3a
C4a
 Biomaterials Research - Manfred Maitz

Methods for Investigation

General/Common pathway
Lysis of sheep red blood cells
Solid phase methods (ELISA, RIA)
Products: C3a, C5a, sC5b-9
Consumption of C3
Ellipsometry
Classical Pathway
Measurement of C1qrs
Measurement of C2b or C4b2a
Alternative Pathway
Measurement of Ba or C3bBb
Measurement of Properidin
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

Biomaterials Consequences
Testing for complement activation included
in standard test programs
Covalent binding of Factor H to the surface
of a blood contacting implant
Reduced C3a, sC5b-9
Inhibition of FXII activation by Heparin-ATIII
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

The Clotting Cascade

Intrinsic Pathway Extrinsic Pathway
Surface Contact Tissue/Cell Defect
Collagen
FXII activator

F XII F XIIa
F VIIa Ca2+ F VII

F XI Ca2+
F XIa

F IX Ca2+
F IXa F III (Tissue
F VIII F VIIIa Thromboplastin)
Platelet Factor 3

Factor F X Ca2+ Factor F Xa Ca2+

Factor F X

F Va FV

Prothrombin I Thrombin
Ca2+

F XIIIa F XIII

Crosslinked Fibrin Fibrin Fibrinogen

Fibrin Meshwork polymers monomers
 Biomaterials Research - Manfred Maitz

The Kinin System

Surface Fibrinolytic
Pathway
Clotting Cascade
F XII F XIIa
(intrinsic pathway) Plasmin
Complement
Activation

Pre-Kallikrein Kallikrein

Plasminogen
Kininogen Kinin
HMWK, LMWK e.g. Bradykinin

Effects
vascular smooth muscle relaxation
Increased permeability of blood vessels Oedema
Pain
Interaction with clotting cascade
Interaction with fibrinolytic cascade
Interaction with complement cascade
 Biomaterials Research - Manfred Maitz

Outline
Complement System
Background
Reactions of the complement cascade
Regulation and activation of the complement cascade
Effects of Complement activation
Evaluation methods
Consequences for biomaterials
Kinin System
Reactions and interactions with other systems
Summary and Conclusions
 Biomaterials Research - Manfred Maitz

Conclusions
The blood plasma has four cascade systems
Clotting cascade
Fibrinolytic cascade
Complement system
Kinin system
All four systems are proteolytic cascades
Besides the main streams there are many cross connections with
minor importance
Common activators and cross activation
Common inhibitors/ regulators
C1 Esterase Inhibitor
Hageman Factor (F XII)
Effective in all systems
Directly activated by surfaces (esp. negatively charged)
1-3% people have deficiencies without clinical syndromes
Plasmin
Directly effective in three systems
Indirectly also effective on the clotting cascade