Part II

Immunology

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 COMPLEMENT

• Complement - System of factors that

occur in normal serum
• Activated by Ag-Ab interaction
• Subsequently mediates a number of
biologically significant consequences

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 COMPLEMENT

• Buchner (1889) – Bactericidal effect of serum

destroyed by heating at 55°C for one hour
• Pfieffer (1894) – Cholera vibrios lysed when
injected into immunised guinea pig
• Bordet and Gengou (1901) – Complement
fixation test

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 COMPLEMENT

• Present in sera of all mammals, animals

birds, amphibians and fish
• Complement - 5% of normal serum protein
• Heat labile, cytolytic activity
• Destroyed at 56°C for 30 minutes

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 COMPLEMENT

• Complement does not bind to free antigen

or antibody but only to antibodies that have
combined with their antigens
• This leads to activation of classical ‘C’
pathway
• All classes of immunoglobulin do not fix
complement - only lgM, lgG3,1 and 2 in
that order.
• Site of ‘C’ binding - Fc fragment of lg
molecule
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 COMPONENTS

• Consists of a least 30 chemically and

immunologically distinct serum proteins
comprising the complement components
• Complement – Nine different fractions
C1-C9
• C1 – Yields three proteins C1 q,r,s
 11 proteins in all
• C1-C9 sequence
Except - C4 comes after C1 and before C2

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 MODEL

• Immune cytolysis of erythrocyte and its

antibody erythrocyte (E) antibody (A)
complex - EA
• (C) component attached = EAC
• Enzymatically activated C1 shown as Ci-
fragment - small letters = C3a, C3b
• Inactivated form = prefix (i-) = (iC3b)

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 COMPLEMENT ACTIVATION

C cascade - Series of reactions in which

preceding component act as enzyme on
succeeding components - cleaving them into
dissimilar fragments

Larger fragment Smaller fragment

 
Joins the cascade Biological effects

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 COMPLEMENT ACTIVATION

 Vascular permeability, smooth muscle

contraction, chemotaxis of leucocytes virus
neutralisation, histamine release, lysis of
cells, virus neutralisation, opsonisation

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 COMPLEMENT PATHWAYS

• Three parallel pathways: Differ only in initial

steps, once C3 activation occurs
-subsequent steps are common to all
pathways
• Classical pathway – First identified - specific
active immunity
• Alternative pathway and lectin pathway –
represent non-specific immunity

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 CLASSICAL PATHWAY
First step - Binding of C1- Ag, Ab (EA)
Recognition unit = C1q - reacts with Fc of lgM
lgG
C1q - six binding sites
Effective activation - C1q attached to
immunoglobulin by at least two of its binding
sites
 1 molecule of lgM or 2 molecules of lgG
initiate/process
C1q binding leads to sequential activate of C1r
and s
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 CLASSICAL PATHWAY

Second step: Ci-s in an esterase


Cleaves several molecules of C4

C4 - split

C4a C4b
 
Anaphylatoxin Binds to cell membrane

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 CLASSICAL PATHWAY

Third step: C4b - presence of Mg - cleaves

• C2-C2a-linked to cell bound C, C4b and
C2b which is released
• C4b2a - Enzymatic activity referred to as C3
convertase

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 CLASSICAL PATHWAY

C3 convertase splits C3 into C3a


Anaphylatoxin
and C3b-cell bound along with

C4b2a-C4b2a3b-enzymatic
called C5 convertase

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 CLASSICAL PATHWAY
Membrane attack - Begins with C5 convertase
cleaving C5 to C5a 
Anaphylatoxin
C5b continues, C6 and C7 join

-C567

Bind to cell membrane

Prepare for lysis by C8 and C9

Cell damage/lysis
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 CLASSICAL COMPLEMENT PATHWAY

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 CLASSICAL COMPLEMENT PATHWAY

The classical pathway of complement

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 ALTERNATE COMPLEMENT PATHWAY

• Central process in complement cascade is

activation of C3 – major component of C
• Activation of C3 with out prior participation
of C4b2a is known as alternate pathway
• Properdin system
• Activator  Bacterial endotoxin
lgA, D
Cobra venom
Nephritic factor

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 ALTERNATE COMPLEMENT PATHWAY
First step - Binding of C3b activator
  factor B
C3b( Bound)

Cleaved by factor D
(C3 proactivator convertase)

Ba Bb
 
Released in medium Bb bound to C3b

C3b Bb  labile
Properdin - factor P
Stabilises C3

Further as in classical

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 LECTIN COMPLEMENT PATHWAY

• Lectins – proteins – bind to carbohydrate

target
• Lectin activates complement – binds to
mannose residues - MB lectin or mannan
binding lectin pathway
• Does not depend on antibody for activation

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 LECTIN COMPLEMENT PATHWAY

• Activated by binding of mannose lectin to

mannose residues present on organisms –
Salmonella, Neisseria and Listeria as well as
Cryptococcus, Candida.
• MBL – Acute phase protein – function similar
to Clq
• Cleavage and activation of C4 and C2-
produces C5 convertase

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 LECTIN PATHWAY

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 OVERVIEW

Overview of pathways of complement activation

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 REGULATION

Inhibitors Inactivators
C1 esterase - heat labile Serum beta globulin
S-Protein - binds to C67 Control of C3
beta globulin factor H
C3 activation
Anaphylatoxin inactivator
Degrades C3a, C4a, C5a
C4 binding protein
controls cell bound C4b

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 BIOLOGICAL EFFECTS

• Cytolysis, bacteriolysis, phagocytosis

• Amplifies inflammatory response
• Hypersensitivity reaction
• Antiviral, phagocytosis
• Interaction with coagulation, fibrinolytic
system

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 BIOLOGICAL EFFECTS

HYPERSENSITIVITY
Cytotoxic-type II, Immune complex type III,
Incompatible transfusion
Thrombocytopenia in sedormid purpura
C-pathogenesis - nephrotoxic nephrits
Immune, complex disease

Serum sickness
Arthus reaction

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 BIOLOGICAL EFFECTS

AUTOIMMUNE DISEASE
C-components decrease in autoimmune

SLE
RA

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 BIOLOGICAL EFFECTS

Endotoxin - Activates alternate pathway


Massive C3 fixation, platelet adherence

Platelet lysis

DIC, thrombocytopenia

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 BIOLOGICAL EFFECTS

IMMUNE ADHERENCE
• Antigen-antibody complex adheres to
erythrocytes – immune adherence -
phagocytosis
CONGLUTINATION
• Bovine serum protein – conglutinin (K) –
clumping of cells coated with C
-conglutination

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 BIOLOGICAL EFFECTS

Biological effects of complement activation

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 DEFICIENCIES OF THE COMPLEMENT SYSTEM

• Complete or partial
• Recurrent bacterial or fungal infection.
• Deficiency of C1 inhibitor - heriditary
angioneurotic edema

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