Completely

central to modern
immunology is
antibody and
immunoglobulin

Sir MacFarlane Burnet 1972

1870 - 1961

Rabbits could be
immunized against RBCs
from another species
resulting in haemolysis
Jules Bordet 1898

Nobel prizes for Medicine

Emil von Behring

1901

Serum therapy

Jules Bordet

1919

Antibody / complement

MacFarlane Burnet / Medawar

1960

Clonal selection theory,

transplantation tolerance

Edelman / Porter

1972

Ig structure

Benacerraf / Dausset / Snell

1980

HLA

Jerne / Kohler / Milstein

1984

Monoclonal antibodies

Tonegawa

1987

Antibody diversity

Murray / Donnall-Thomas

1990

BMT

Doherty / Zinkernagal

1996

HLA

Steinman

2011

DCs - HLA

Synopsis
Basic structure
Ig domains/superfamily
Ig classes

Structural diversity
VDJ recombination
somatic hypermutation

Structure-function correlations
Ig classes and normal function
disease

Uses

IMMUNOGLOBULIN (Ig)
SUPERFAMILY

Ig domain/fold
70-110 aa
self- stabilising/
folding unit via
disulphide bridges
between conserved
cysteine residues
forms a barrel-like
shape

SANDWICH OF
TWO -SHEETS

Bork P, 1994

Immunoglobulin superfamily
evolution
expressed in jawed vertebrates from
500 million years ago
one third of cell surface receptors

CELL-CELL
INTERACTION
self & non-self
IMMUNE RECOGNITION
CELL-ADHESION - ORGANOGENESIS

neural cell adhesion molecule

CD56 homophilic binding

Molecule function/category

Examples

Antigen receptors

ANTIBODIES/ IMMUNOGLOBULINS; TCR

Antigen presenting molecules

MHC I; MHC II;

Co-receptors

CD4; CD8; CD19

Antigen receptor accessory

molecules

CD3-, - and - chains; CD79a; CD79b

Co-stimulatory or inhibitory
molecules

CD28; CD80; CD86

Receptors on Natural killer cells

Killer-cell immunoglobulin-like receptors

(KIR)

Adhesion molecules

Neural Cell Adhesion Molecule (NCAM/

CD56); CD2; SIGLEC family (CD22, CD83);
CTX family (CTX, JAMs, BT-IgSF, CAR, VSIG,
ESAM); Intercellular adhesion molecules
(ICAMs); vascular cell adhesion molecules (
VCAM-1)

Cytokine and growth factor

receptors

IL-1R; PDGFR; IL-6R; M-CSF-R; c-kit

Others

thymocyte differentiation antigen-1 (Thy-1);

sodium channel subunit beta-1 precursor

Fab combines variable regions of

heavy and light chains Ag binding 2

identical heavy chains

(isotypes:

Subclasses have different

glycosylation & disulphide bridges
due to allelic variation mainly on the
heavy chain (allotypes)
2 identical light chains (

Fc crystalizable / constant
Fragment binds C and immune cells

 PROTOTYPE, default Ig
first Ig produced in response
to antigen challenge
also exists as a membrane
bound receptor on nave B
cells
in serum forms a pentamer
powerful fixer of complement

Why is IgM first?

Why switch?

AFFINITY:
VALENCY:
AVIDITY:

strength of the binding between a single paratope of

an antibody and a single antigenic epitope
IgM low affinity, IgG high affinity
number of epitopes
IgM - decavalent (10), IgG divalent (2)
total affinity of an antibody with more than
one binding site

the predominant Ig in serum

in humans there are 4
subclasses
subclasses vary in disulphide
bonds and glycosylation sites
IgG is the only Ig to cross the
placenta
arises from a mechanism
known as class switching
from
IgM producing activated
B-cells

Notarangelo LD et al, 2006

Hyper IgM syndrome

high IgM, low or absent IgG and IgA

absent specific antibody production
risk of bacterial infections
treat with immunoglobulin replacement
(BMT for X-linked form)
Davies & Thrasher, Br J Dermatol, 2010

serum IgA has a similar

monomeric structure to IgG
in secretions it forms a dimer
its heavy chain is slightly larger at
60K
two subclasses: IgA1 and IgA2
as with IgM, polymerisation is
initiated and stabilised by J chain
the predominant isotype in
breast milk
important for mucosal immunity?

IgM

IgG

IgA

IgE

+++

++

Binds to neutrophils

+++

Binds to mast cells/

basophils

+++

Crosses the placenta

++

Present in breast milk

++

Complement
activation

Immunoglobulin
diversity
Ig required for recognition of myriad of Ags
10 billion different antibodies

number of genes limited

30 V genes, 300 V genes, 1,000 heavy V genes

somatic mutation - VDJ recombination

somatic hypermutation - point mutations within
hypervariable regions
Schroeder & Cavacini, JACI, 2010

VDJ recombination

Hypervariable regions
Complementarity determining
Regions (CDR)
antigen binding specificity

Cytosine

activation-induced
(cytidine) deaminase
(AID)

Uracil DNA glycosylase

Uracil

Kavli B et al, 2007; Neuberger & Rada, JEM, 2006

B-cell
clonal
selection

ANTIBODY PREVENT INFECTION BY

PROMOTING
THE UPTAKE OF ENCAPSULATED
BACTERIA BY PHAGOCYTES

OPSONISATION

ANTIBODY PREVENT INFECTION BY

BLOCKING
THE UPTAKE OF BACTERIAL TOXINS
AND VIRUSES BY TARGET CELLS

Mechanism of action
TOXIN

VIRUS

ANTIBODY

ACTIVATES
CLASSICAL COMPLEMENT CASCADE

opsonisation
membrane attack complex

Nephelometry
Gk nephele = cloud

measures Igs,
C components
depends on light scatter
properties of Ag/Ab complexes
add antisera of component to
be measured complexes
turbidity
check absorbance at 840nm
Advantages
High throughput
Rapid, automated
Disadvantages
High initial set up cost
Interference if turbid sera/lipemia/
Freeze thawing

Immunoglobulin (IVIG)
Therapeutic preparation of normal
human IgG obtained from pools of
plasma from 3,000 to 100,000 healthy
blood donors (Cohn et al, 1946)
Diverse repertoire of IgG molecules
that possess a wide spectrum of
antibacterial and antiviral specificities
Specificity may vary depending on
population from which it is drawn

Immunoglobulin therapy
Replacement therapy IV or SC
1o 2o immunodeficiencies

Passive immunisation
e.g. tetanus, rabies, hepatitis A&B, VZV, RSV

Monoclonal antibody therapy

selective cytotoxic: e.g. B-cells rituximab, T-cells anti-CD3, cancers
anti-inflammatory: e.g. TNF infliximab, IL-1 anakinra

Laboratory uses
monoclonal antibodies
Identification/localisation of cellular
protein
FACS
immunoflurescence
immunohistochemistry

Identification of soluble proteins

immunoprecipitation
Western blot
ELISA

Summary
Immunoglobulins are critical in biology
cell adhesion self to self recognition
Immunity self to non-self recognition

Diversity is generated by enzymes which

cut and paste DNA
defects are associated with propensity to
infections (immunodeficiency) and to cancer

Immunoglobulins are used widely in

laboratory and clinical medicine
monoclonal antibodies form the basis of
ELISA, flow cytometry, biologic therapies