P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement
Definition : series of heat-labile serum proteins Site : serum and all tissue fluids except urine and CSF

Synthesis : in liver appear in fetal circulation during 1st 13 W Function : Responsible for certain aspects of immune response and inflammatory response

Activation : antigen-antibody complex or endotoxin, capsule series of proteins activated sequentially Inactivation: inhibitors in plasma (short lived)

Biological effects: either beneficial or harmful to host P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement: History
Discovered in 1894 by Bordet
It represents lytic activity of fresh serum Its lytic activity is destroyed when heated at 56C for 30 min
P G UPADHYAYA, KIMSMICRO,BANGALORE

Properties
Approximately 30 circulatory & membrane bound proteins complement refers to ability of these proteins to complement, i.e., augment, the effects of other components of immune system Important component of innate host defenses Synthesized in the liver and by cells involved inP G UPADHYAYA, KIMSMICRO,BANGALORE the inflammatory response.

Effects
Lysis : of cells such as bacteria, allografts, and tumor cells. Generation of mediators that participate in inflammation and attract neutrophils Opsonization, ie, enhancement of phagocytosis

P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement Functions

Host detriment: Inflammation Anaphylaxis

Figure-1. Complement has a central role in inflammation as it causes chemotaxis of phagocytes, opsonization and lysis of pathogens and clearence of immune P complexes. G UPADHYAYA, KIMSMICRO,BANGALORE

Proteins of the complement system (nomenclature)

C1(qrs), C2, C3, C4, C5, C6, C7, C8, C9 factors B, D, H and I, properdin (P) mannose binding lectin (MBL), MBL associated serine proteases (MASP-1 MASP-2) C1 inhibitor (C1-INH, serpin), C4-binding protein (C4-BP), decay accelerating factor (DAF), Complement receptor 1 (CR1), proteinS (vitronectin)
P G UPADHYAYA, KIMSMICRO,BANGALORE

Definitions
C-activation: alteration of C proteins such that they
interact with the next component

C-fixation: utilization of C by Ag-Ab complexes Hemolytic units (CH50): dilution of serum which
lyses 50% of a standardized suspension of Ab-coated r.b.c.

C-inactivation: denaturation (usually by heat) of an

early C-component resulting in loss of hemolytic activity

Convertase/esterase: altered C-protein which acts

as a proteolyticUPADHYAYA, KIMSMICRO,BANGALORE enzyme for another C-component PG

Activation

several complements are pro-enzymes, which must be cleaved to form active enzymes.

Initiated either by antigen-antibody complexes or by a variety of non-immunologic molecules, eg, endotoxin

P G UPADHYAYA, KIMSMICRO,BANGALORE

Activation product of complement proteins (nomenclature)

Activated component are usually over-lined: e.g. C1qrs When enzymatically cleaved, the larger moiety, binds to the activation complex or membrane and the smaller peptide is released in the microenvironment Letter b is usually added to the larger, membrane-binding, peptide and a to the smaller peptide (e.g., C3b/C3a, C4b/C4a, C5b/C5a), EXCEPT C2 (the larger, membranebinding moiety is C2a; the smaller one is C2b)
P G UPADHYAYA, KIMSMICRO,BANGALORE

Pathways of complement activation

CLASSICAL PATHWAY
antibody dependent

LECTIN PATHWAY

ALTERNATIVE PATHWAY

antibody independent

Activation of C3 and
generation of C5 convertase

activation of C5
LYTIC ATTACK PATHWAY
P G UPADHYAYA, KIMSMICRO,BANGALORE

Importance

Alternative pathway is more important the first time we are infected by a microorganisms, because the antibody required to trigger the classic pathway is not present. Both pathways lead to the production of C3b, the central molecule of the complement cascade. C3b has 2 important functions: (1) It combines with other complement components to generate C5
P G UPADHYAYA, KIMSMICRO,BANGALORE

Importance of C3b
(1) It combines with other complement components to generate C5 convertase, the enzyme that leads to production of MAC.
(2) Opsonizes bacteria because phagocytes have receptors for C3b on surfaces

P G UPADHYAYA, KIMSMICRO,BANGALORE

Components of the Classical Pathway

C3

C4

C1 complex
C1s is an enzyme and cleaves C4 and C2
P G UPADHYAYA, KIMSMICRO,BANGALORE

Classical Pathway Generation of C3-convertase

Cleavage of C4 by C1s produces C4a and C4b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Classical Pathway Generation of C3-convertase

C4b binds C2 and C4b2 is cleaved by C1s. C2b is released but C2a remains bound to C4b on the surface. C4b2a is C3 Convertase

C4b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Classical Pathway Generation of C5-convertase

________ C4b2a3b is C5 convertase; it leads into the Membrane Attack Pathway

C4b

C3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Biological Activities of Classical Pathway Components

Component
C2b C3a

Biological Activity
Prokinin; cleaved by plasmin to yield kinin, which results in edema Anaphylotoxin; can activate basophils and mast cells to degranulate resulting in increased vascular permeability and contraction of smooth muscle cells, which may lead to anaphylaxis Opsonin Activation of phagocytic cells Anaphylotoxin Opsonin
P G UPADHYAYA, KIMSMICRO,BANGALORE

C3b C4a C4b

Control of Classical Pathway Components

Component
All C3a C3b C4a C4b

Regulation
C1-inhibitor (C1-INH); dissociates C1r and C1s from C1q C3a-inactivator (C3a-INA; Carboxypeptidase B) Factors H and I; Factor H facilitates the degradation of C3b by Factor I C3a-INH C4 binding protein (C4-BP) and Factor I; C4-BP facilitates degradation of C4b by Factor I; C4-BP also prevents the association of C2a with C4b thus blocking formation of C3 convertase
P G UPADHYAYA, KIMSMICRO,BANGALORE

C1-inhibitor deficiency: hereditary angioedema

P G UPADHYAYA, KIMSMICRO,BANGALORE

Components of mannose-binding lectin pathway

Pathogen

MBL

MASP1

P G UPADHYAYA, KIMSMICRO,BANGALORE

Mannose-binding lectin pathway

_____ C4b2a is C3 convertase; it will lead to the generation of C5 convertase
MASP1

MBL

Binding to lectins cause autocatalytic activation of MASPs which then cleave C4 & C2
P G UPADHYAYA, KIMSMICRO,BANGALORE

Components of the alternative pathway

C3

P G UPADHYAYA, KIMSMICRO,BANGALORE

Spontaneous C3 activation

Generation of C3 convertase

C3

C3b

fB activate fD which then cut fB releasing Ba, while Bb becomes an active protease C3Bb complex has a very short half life P G UPADHYAYA, KIMSMICRO,BANGALORE

C3-activation the amplification loop If spontaneously-generated C3b is not degraded

C3b

C3 b

P G UPADHYAYA, KIMSMICRO,BANGALORE

C3-activation the amplification loop

C3 b

C3b

C3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

C3-activation the amplification loop

C3b

C3b

C3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Control of spontaneous C3 activation via DAF

DAF prevents
C3b

the binding of

factor B to C3b

CR1 Autologous cell membrane

P G UPADHYAYA, KIMSMICRO,BANGALORE

Control of spontaneous C3 activation via DAF

DAF dislodges

C3b-bound
factor Bb

C3b

CR1 Autologous cell membrane

P G UPADHYAYA, KIMSMICRO,BANGALORE

Control of spontaneous C3 activation via CR1

C3b

C3b

CR1 Autologous cell membrane

P G UPADHYAYA, KIMSMICRO,BANGALORE

iC3b CR1

Degradation of spontaneously produced C3b

C3c

C3c

C3b
C3dg iC3b

C3b
C3dg iC3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

C3b stabilization and C5 activation

C3b finds an activator (protector) membrane
This is stable C5 convertase of the alternative pathway C3b b

C3 b

P G UPADHYAYA, KIMSMICRO,BANGALORE

C3b regulation on self and activator surfaces

C3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

C5-convertase of the two pathways

C5-convertase of the Classical and lectin Pathways C5-convertase of the Alternative Pathway

C3b

C3b

C3b

C4b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Lytic pathway

Generation of C5 convertase leads to the activation of the

Lytic pathway
P G UPADHYAYA, KIMSMICRO,BANGALORE

Components of the lytic pathway

C7 C6

C 9

P G UPADHYAYA, KIMSMICRO,BANGALORE

Lytic pathway C5-activation

b C4b C3b

P G UPADHYAYA, KIMSMICRO,BANGALORE

Lytic pathway assembly of the lytic complex

C5b first binds C6 and then C7 from the plasma. Membrane bound

C6 b

C5b67 recruits C8 and C9 to form

the Membrane Attack Complex (MAC)

C7

P G UPADHYAYA, KIMSMICRO,BANGALORE

Lytic pathway:
insertion of lytic complex into cell membrane

C6 b

C7 CC C C C9 9 9 9C 9C C C9 9 9 9
P G UPADHYAYA, KIMSMICRO,BANGALORE

Biological effects of C5a

P G UPADHYAYA, KIMSMICRO,BANGALORE

Biological properties of C-activation products

Product
C2b (prokinin) C3a (anaphylatoxin)

Biological Effects
edema

Regulation
C1-INH

mast cell degranulation; carboxypeptidase- B enhanced vascular (C3-INA) permeability; anaphylaxis

P G UPADHYAYA, KIMSMICRO,BANGALORE

Biological properties of C-activation products

Product
C3b (opsonin)

Biological Effects
opsonization; phagocyte activation

Regulation
factors H & I

C4a as C3, but less (anaphylatoxin) potent

(C3-INA) C4-BP, factor I

C4b (opsonin)

opsonization; phagocytosis

P G UPADHYAYA, KIMSMICRO,BANGALORE

Biological properties of C-activation products

Product
C5a (chemotactic factor)

Biological Effects
anaphylactic as C3, but much more potent; attracts & activates PMN causes neutrophil aggregation, stimulation of oxidative metabolism and leukotriene release chemotaxis, attaches to other membranes
P G UPADHYAYA, KIMSMICRO,BANGALORE

Regulation
carboxypeptidase-B (C3-INA)

C5b67

protein-S

Complement Deficiencies and Disease Classical Pathway

Pathway Component C1INH Disease Hereditary Angioedema Predisposition to SLE Mechanism Overproduction of C2b (prokinin) Opsonization of immune complexes help keep them soluble, deficiency results in increased precipitation in tissues and inflammation

C1, C2, C4

P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement Deficiencies and Disease Lectin Pathway

Pathway Component MBL

Disease Susceptibility to bacterial infections in infants or immunosuppressed

Mechanism Inability to initiate lectin pathway

P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement Deficiencies and Disease Alternative Pathway

Pathway/Component Factors B or D Disease Susceptibility to pyogenic (pus-forming) bacterial infections Susceptibility to bacterial infections Susceptibility to Gramnegative infections Mechanism Lack of sufficient opsonization of bacteria

C3

Lack of opsonization and inability to utilize the membrane attack pathway Inability to attack the outer membrane of Gramnegative bacteria

C5, C6, C7 C8, or C9

P G UPADHYAYA, KIMSMICRO,BANGALORE

Complement Deficiencies and Disease Alternative Pathway cont.

Pathway Component Properdin (X-linked) Disease Susceptibility meningococcal meningitis Mechanism Lack of opsonization of bacteria

Factors H or I

C3 deficiency and susceptibility to bacterial infections

Uncontrolled activation of C3 via alternative pathway resulting in depletion of C3

P G UPADHYAYA, KIMSMICRO,BANGALORE