Dr C G Lopez

Ex Transfusion Medicine Unit

University Malaya Medical Centre
Innate Non Specific
Immune Response
Innate immune defenses
Non-specific

This system does not confer long-lasting

immunity against a pathogen.

The innate immune system is the dominant

system of host defense
Innate immune defenses
Involves the following:
phagocytic cells (neutrophils, monocytes,
and macrophages)
cells that release inflammatory mediators
(basophils, mast cells, and eosinophils)
natural killer cells (NK cells)
complement proteins
 cytokines
 Innate immune defenses

anatomical barriers

 mechanical removal

bacterial antagonism

phagocytosis

pattern-recognition receptors,

antigen-nonspecific defense chemicals,

 complement pathways
Innate Immunity
Designed to recognize molecules ( PAMPS) shared by groups
of related microbes. Pathogen-Associated Molecular Patterns
( PAMPS ) include
- LPS from the gram-negative cell wall,
- peptidoglycan and lipotechoic acids from the gram-
positive cell wall
- bacterial and viral unmethylated DNA
- bacterial flagellin,
 the amino acid N-formylmethionine found in bacterial proteins,
double-stranded and single-stranded RNA from viruses
glucans from fungal cell walls.
unique molecules displayed on stressed, injured, infected, or
transformed human cells also act as PAMPS.
Innate Response
Response triggered when

• microbes are identified by pattern

recognition receptors common to broad
groups of microorganisms

• damaged, injured or stressed cells send out

alarm signals, many of which (but not all) are
recognized by the same receptors as those
that recognize pathogens.
Immediate Innate
Response

njured or infected cells release eicosanoids

and cytokines

rostaglandins inflammation ( redness and

swelling )

eicosanoids)

ytokines

eucotrenes attract WBC ‘s to site of

 Eosinophi
T
ls cells

Bcells

Kille
r
cells
PLASM
A
CELLS Neutrophil
s
MACROPHAG
E
Neutrophils
Most abundant type of leukocytes in body
Highly effective in killing most bacteria and fungi.
PMNs contain many cytotoxic compounds that are non-
specific – needs to be regulated
PMN turnover regulated by apoptosis, a process of cell
death and safe removal by macrophages.
Apoptosis is accelerated following phagocytosis of
bacteria, a process that appears important for the
resolution of infection and inflammation.
neutrophils which have receptors for C3b
Specific Immune
Response
Cluster of genes MHC
or HLA region –
encode HLA antigens ,
complement
components
MHC Class l Antigens
Class I molecules expressed by almost all the cells
of the body .
Surface heterodimers that primarily present
peptides derived from the cytosol (viral bacteria,
pollen, self peptides) to circulating CD8+ T cells.

Act as ligands for killer immunoglobulin receptors

(KIR), which regulate the cytotoxic activity of
natural killer (NK) cells.

Epitopes that are part of class I histocompatibility

molecules bind to CD8+ T cells
Class II molecules
Class II molecules - Only specialized antigen-
presenting cells express these i.e dendritic
cells phagocytic cells like macrophages and B
cells

alpha-beta heterodimer presents primarily

exogenously derived peptides (bacteria and
chemical toxins) to circulating CD4+ T cells.

Epitopes that are part of class II

histocompatibility molecules bound to CD4+
 CLASS I and II
Alpha and beta
chains encoded HLA Antigens
by class ll genes Alpha chain
encoded by class 1
genes

Encoded by gene on
Chr 15 . May be
concerned with trans
membrane signaling

B “activated” T On almost all nucleated

lymphocytes, dendritic cells and platelets
lymphocytes, macrophages Soluble HLA antigens
found in plasma
 CD28
Stimulation through CD28 in addition to
the TCR provides a potent co-stimulatory
signal to T cells for the production of
various interleukins (IL-2 and IL-6 in
particular).
 Cytotoxic T cell Class 1
Pathway

CD8+ T cells bind epitopes

part of class I
histocompatibility molecules )
.
Almost all the cells of the
body express class I
molecules.
T 4 Class II
Pathway
The alpha-beta
heterodimer presents
primarily exogenously
derived peptides
(bacteria and
chemical toxins) to
circulating CD4+ T
cells.

CD4+ T cells
bind epitopes
part of class ll
histocompatibility
molecules
NK cells
Unlike B and T cells, NK cells do not express
unique clonally distributed receptors for specific
antigens, rather they express many different
promiscuous stimulatory and inhibitory receptors
that can be divided into at least four
the killer immunoglobulin-like receptors (KIRs)
 the C-type lectin receptors
 the natural cytotoxicity receptors (NCRs)
toll-like receptors (TLRs)
 NK cells: KIR receptors
(KIRs) are Killer cell immunoglobulin-like receptors for
classical MHC I (HLA-A, HLA-B, HLA-C) molecules. Some
KIRs are specific for certain HLA subtypes

Provides the first line of defense against virus infection and

tumor transformation.

KIRs - interacts with MHC class I molecules

- regulate the cytotoxic activity of natural killer (NK)
cells
- distinguish the tumor and virus infected
cells from normal body cells.
.
Specific Non clonal
Clonal Receptors
Receptors
Large
granular NK Cells bind
Killer Cells tumour targets
with no T or B
markers

Killer Cells have

Fc receptors . Can
bind Ab already
bound to tumour NK Cells bind targets
target by unknown mechanism
Probably perforin . Kill cells thru perforin
release mechanism release mechanism
kills cells
B cells
• B lymphocytes are both antigen-receiving and antigen-
presenting cells.
• They can come in contact with these antigens by
- encountering them in the surrounding lymph
- being presented them by macrophages or
dendritic cells.
They bind intact antigens (e.g., virus particles, proteins)
with their B cell receptor (BCR).

• B lymphocytes process antigen by the class II

pathway for presentation to T cells.
 Macrophage with Ag
presented
to T helper lymphocyte B CELL
B CELL Activated by cytokines

More Helper T cells

More Cytotoxic T clls
( CTLs)
B cells to Plasma cells
The Complement System
 Characteristics of
•Series of proteins found in fresh, normal serum.
Complement
• Categorized as a beta globulin.
• Found in the beta region on protein electrophoresis
• C3 is complement component that is found in the
highest
concentration

Complement components are identified by C and

their number: C1, C2, C3 etc
Complement products resulting from the splitting of
these proteins during the activation process are
followed by a lower case letter: C3a, C3b, C1q
If complement complexes develop that have
enzymatic activity are written with a bar above the
top: C5b678
Complement controls various biological processes

• Cell destruction through lysis

• Cell destruction through opsonization (enhanced
phagocytosis) especially with C3b
• Chemotaxis via certain split products acting as
chemical signals to the phagocytic cells
• Anaphylaxis again through the split products (C5a and
C3a), which promote inflammation. C5a and C3a
can bind with mast cells and basophils leading to
the release of histamine. This is turn:
• Increases vascular permeability
• Smooth muscle contraction to preserve blood for vital
organs
• Increases cellular membrane adhesion
Complement and Innate Immune Response
Complement proteins bind to
- carbohydrates on the surfaces of microbes ( C4b)
- antibodies attached to these microbes or cells
Complement protease activity activated producing a
catalytic cascade that amplifies the initial signal
Cascade results in production of peptides - attract immune
cells, increase vascular permeability, and opsonize (coat)
pathogen surface marking it for destruction.
Complement can kill cells directly by disrupting their
plasma membrane
Complement Activation
Comprises the :-

Classical pathway
Activated by antigen-antibody complexes and may
require all nine complement proteins.

Lectin pathway
Initiated by bacterial surface sugars (mannose) through the
mannose-binding lectin (MBL) protein and subsequent interaction
with mannose-binding lectin-activated serine proteases.

After that point pathway functions identically to classical pathway.

Alternative pathway
activated by LPS.
Alternative Pathway

Activated by C3b bound

to surface components
of microbes

MASP: MBL-
associated
serine
proteinase
MBL: mannan-
binding lectin;
MCP:
Proteins of the Classical pathway
CI
in plasma as complex of - 6 mol CIq, 2 mol CIr ,
2 mol Cls

Binding of CIq - activates CIr , Cls

Activated Cl (serine protease ) cleaves C4 into

large fragment C4b and small C4 fragment
which diffuses away

C4b binds covalently to sugar residues on cell

surface glycoproteins
C2

Cleaved to C2b ( C2a diffuses away ) which

binds non- covalently to the large fragment C4b

Complex C4b*2a = “C3 Convertase”

 C3 Most abundant protein - 1.3 mg/ml
Cleaved by “C3 Convertase” (called C4b* 2a before ) to

C3b
• binds to covalently to
glycoproteins on phagocytes–e.g. C3a diffuses into
macrophages, neutrophils which plasma can bind to
have receptors for C3b. basophils, mast cells
which release
• C3b coats particles e.g bacteria vasoactive contents –
before they are phagocytosed - histamine - acts as
acts as opsonin anaphylotoxin )

• binds to C5 to form C3/ C5

Convertase
C5

Cleavage of C5 by “C3/C5 Convertase” produces

set of complement proteins for membrane attack –
called Membrane Attack Complex

C5a released into surrounding fluid – acts as

anaphylatoxin ( like C3a ) and chemotactic
attractant for neutrophils

C5b serves as anchor for assembly of single

molecule each of C6, C7, C8.
Resulting complex C5b *6*7*8 guides C9
polymerisation
As many as 18 molecules of C9 form a
channel into the bi lipid layer of the cell
membrane allowing passage of ions and
small molecules

Water enters cell by osmosis and cell lysis

Note: Membrane Attack Complex by another C5

convertase produced by the Alternate Pathway
C5a anaphylatoxin
Contains 74 amino acids

Induces
• release of granular enzymes from
phagocytic cells
• production in neutrophils of superoxide
anion
• vasodilatation
• increased vascular permeability
• induction of thymocyte apoptosis during
sepsis

Excessive production of C5a by activation

in sepsis can lead to an unregulated pro-
inflammatory response, ultimately
resulting in tissue damage and multi-
 The Alternate ( Properdin ) Pathway

Does not require specific antibody to recognise potential

pathogens.

6 plasma proteins perform – A, (C3), B (C3 proactivator ),

D, H, I, perform continuous surveillance function

Activated by
- Organisms - bacteria, fungi, certain viruses
- virus infected cells
- variety of polysaccharides & lipopolysaccharides
- human RBC lacking Decay Accelerating Factor
 The Alternate ( Properdin ) Pathway

C3b continuously generated in circulation in small

amounts formed from intervention with activators e.g cell
walls of bacteria, fungi, ect also endotoxins

C3b + Factor B ( C3 Proactivator ) forms complex

Complex C3bBbP ( activated Factor B )
Activates large numbers of C3 molecules
Alternative Pathway

Activated by C3b bound

to surface components of
microbes ( opsonised )

MASP: MBL-
associated
serine
proteinase
MBL: mannan-
binding lectin;
MCP:
Proteins of the MBL ( Mannose Binding Lectin )
Pathway
MBL made in liver in response to Macrophage
cytokines

MBL cascade initiated by MBL binding to

pathogen surface ( microbial carbohydrates)

MBL and 2 serum proteases C4 & C2 function

like CI to form C3 convertase

C3b combines with C5 & C6 to form C5

convertase
All pathways converge at level of C3
leading to

• the cleavage products, C3a and C5a

• terminal membrane attack complex,

C5b-9, which forms pores in the
membranes of bacteria and cells,
ultimately causing their lysis.
Summary of Effector Functions of Complement

Opsonization by C3b – targets foreign particles for

phagocytosis

Chemotaxis by C5 a attracts phagocytic cells to site

of damage – aided by increased permeability of the
capillary beds mediated by C3a and C5a

Assists in catabolism of Ag/Ab complexes and

elimination from body Deficiency of C2 associated
with Lupus Erythematosis – an auto immune disease

Lysis of antibody coated cells

Immunoglobulins
Each
antibody
molecule
has specific
Ag binding
site
                
           
    

       
 
                                
Macrophage
receptors bind
here

                                                 
 Immunoglobin Molecule and Complement

CIq – largest six globed structure

fused by shafts to single base

Serves as recognition unit that

binds to Fc region of Immunoglobin
molecule
to Ch2 domain - if IgG
to Ch 4 domain - if IgM
Immunoglobin Molecules and Complement
IgG less efficient at complement binding than
IgM

Two IgG molecules with FcR binding sites

must be in close proximity for CIq to attach and
activate the full complement cascade

IgM has 5 Fc sites. One molecule of IgM is

capable of CIq binding to activate the full
complement cascade
 One IgM
molecule
able to span
distance
between cells
and bind CIq

CI
Two IgG
molecules
required
to bind CIq
 IgM antibodies able to
span the distance and
bring about direct
agglutination

IgG antibodies unable to span

the distance cells are
sensitized but agglutination
not seen