2020/5/7 1
Immunoglobulins
Jun Dou (窦骏)
Department of Pathogenic biology and Immunology
School of Medicine, Southeast University
May 8,2020
2020/5/7 2
Immunoglobulin (Ig) are the critical
ingredients at every stage of a humoral
acquired immune response. When expressed
on the surfaces of resting B lymphocytes,
they serve as receptors that can detect and
distinguish among the vast array of potential
antigens present in an environment.
2020/5/7 3
The Ig that secreted as a result then
function as antibodies, traveling through
the tissue fluids to seek out and bind to the
specific antigens that triggered their
production.
2020/5/7 5
Antibody structure
•Two heavy chains
•Two light chains
*H chain---50KD
* L chain---25KD
2020/5/7 6
Antibody structure?
V region (domain)Regions vary greatly in
amino acid sequence in
different Abs, involved
in Ag recognition.
C region (domain)relatively constant in
sequence, engaging
the effector functions.
2020/5/7 7
amino terminus
carboxyl terminus
glucoprotein
2020/5/7 8secondary structure
2020/5/7 10
tertiary structure
Complementarity
Determining Region
2020/5/7 11
Complementarity
Determining Region
2020/5/7 12
Antibody structure
hydrolysis
2020/5/7 15
Antigen Antibody
Interactions
2020/5/7 17
2020/5/7 18
The Sheet
Anti-parallel βSheet
2020/5/7 19
2020/5/7 20
2020/5/7 21
2020/5/7 22
Immunoglobulin Supergene Family.
2020/5/7 23
monomer
2020/5/7 25
Generation of Secretory Immunoglobulins
IgA
dimer
2020/5/7 26
2020/5/7 27
2020/5/7 28
Immunoglobulin Subtypes
2020/5/7 29
Functions of Antibody
◼ Neutralize toxins
◼ Bind to bacterial surfaces and aid in
opsonization
◼ Bind to bacterial surfaces and aid in
complement killing
◼ Inactivate viral particles
◼ Provide immunity to the unborn
◼ Provide immunity to the newborn
2020/5/7 30
Functions of Antibody
•Neutralization
•Opsonization
•Complement Activation
2020/5/7 33
Neutralization toxicant
2020/5/7 34
2020/5/7 35
2020/5/7 36
Immunoglobulin Isotypes
◼ Isotypes are the antigenic features of a class
of immunoglobulin H (heavy chain).
◼ Example for µ chain is isotypical different
from chain.
◼ The genes for 1, 2, 3, 4, µ, 1, 2, ,Ɛ,
chains are present in the human genome, and
are therefore isotypes.
2020/5/7 37
◼ IgM Big, First, Complement
◼ IgD Membrane receptor
◼ IgG Secondary response, prototype
◼ IgA Secretions
◼ IgE Allergy (Wheeze)
2020/5/7 38
Isotype
Allotype
Idiotype
2020/5/7 39
◼ Allotype
◼ Allotypes are additional features of immuno-
globulines that vary among individuals
within a species and are under genetic
control.
◼ Allotypes can be shared by some members of
a species.
2020/5/7 40
◼ Idiotype
◼ The term idiotype refers to the unique V-
region amino acid sequences of the
homogeneous immunoglobulin molecules
produced by a single B cell clone.
◼ Thus, there are as many idiotypes as there are
B cell clones (perhaps about 108 in an adult).
It is now known that anti-idiotype Ab
specificity recognize sequences in the
hypervariable regions of the target Ab that
induced it.
2020/5/7 41
An anti-idiotype immune response (Ab or T
cell mediated) expresses its own idiotype
which in turn can be recognized as foreign
and an anti-idiotype immune response made
against this idiotype.
Jerne described a Network Theory which
proposes that a series of idiotype-anti-idiotype
reactions are partially responsible for
regulation of the immune response.
2020/5/7 42
How does the immune system
make millions or billions of different antibody molecules?
2020/5/7 43
Mechanisms for Generation of Diversity
◼ Random assortment of light and heavy chains.
◼ Multiple V region gene segments.
◼ Somatic rearrangement.
◼ Faulty joining.
◼ Somatic mutation of rearranged genes.
2020/5/7 44
How to determine Ig gene structure?
Genetic structure (DNA) Ab (peptide)
2020/5/7 45
How are Ig genes rearranged?
**H chain genes are rearranged first, followed by L chain genes
2020/5/7 46
Somatic Rearrangement of Ig genes occurs during B cell differentiation
Ig M IgD
proteins
(Heavy chains)
VH >100 DH 1-12 JH 1-14
V, D, J Rearrangement. If 100 V segments and 12 D
segments and 14 J segments. 100x12x14 = 16800
different combinations are possible.
C C C 3 C 2 C 1 C 2 C 4 C 1 C 2
Transcription
Translation
mRNA
2020/5/7 47
TCR genes construct and rearrangment
Southern blot analysis
EcoRI digesting PCR products
TCR— and genes use a strategy of
recombination similar to that of Ig genes.
2020/5/7 48
Primary and Secondary Responses. Specific Adaptive or Acquired Immunity.
2020/5/7 49
Primary and Secondary Immune Response to Antigen
Antigen A and Antigen BAntigen A
Primary
response
to A
Primary
response to B
Secondary response to A
Time
[Antibody]
New Types
of Antibodies
IgG
IgA
IgE
2020/5/7 50
Antibody Use in Medicine
◼ Detecting assays
◼ Drugs, chemicals, hormones, pregnancy
tests, HIV tests, etc
◼ Enzyme linked immunosorbent assay
(ELISA)
◼ Blood typing
◼ Microbial identification
2020/5/7 51
◼ Passive Antibody Therapy.
◼ e.g. Gamma globulins
◼ Imaging
◼ Immunotoxins. Cancer therapy
◼ Neutralize inflammatory cytokines. e.g.
antibody to TNF
◼ Catalytic antibodies.
◼ Etc.
2020/5/7 52
ELISA Western
Blot
2020/5/7 53
2020/5/7 54
Polyclonal antibody Monoclonal antibody (McAb)
2020/5/7 55
Monoclonal
Antibody
2020/5/7 56
Blood typing by agglutination
2020/5/7 57
Immunoprecipitation
2020/5/7 58
Immunodiffusion
2020/5/7 59
Double
Immunodiffusion
2020/5/7 60
2020/5/7 61
Immunoflourescence
2020/5/7 62
2020/5/7 63
FACSOr
Flow
Cytometry
2020/5/7 64
Humoral Immunity In Summary:
◼ Antibody functions by binding antigen and by
directing the activity of complement and
phagocytes
◼ Antibody is a protein termed immunoglobulin
◼ There are several different Immunuglobulin
classes or isotypes depending on the heavy chain
expressed.
2020/5/7 65
◼ Antibodies can exist in millions or billions
of different forms (antigen specificities)
created via a process termed generation of
diversity.
◼ Antibody production results from B cell
proliferation of differentiation.
◼ B cell proliferation and differentiation
usually requires helper T cells.
2020/5/7 66
◼ Extracurricular works I. Please answer the concepts below:
◼ 1. Ig and Antibody(Ab)
◼ 2. Ig Isotype
◼ 3. Ig Allotype
4. Ig idotype
◼ 5. Monoclonal Ab and Polyclonal Ab
• 6. Neutralization
◼ 7. Opsonization
◼ 8. CDRs and VH
◼ 9. Fab and Fc
◼ 10. Noncovalent forces
◼ 11. Ig subtypes and sIgA
2020/5/7 67
◼ II. Please answer the questions below:
◼ 1. Please demonstrate the basic structure of
immunoglobulin.
◼ 2. What are the functions of antibodies?
◼ 3. What are the differences of primary and
secondary response for antibody production?
◼ 4. How to understand the passive Ab therapy?
◼ (hint: COVID-19 therapy)