10/21/2014

We already know
B cells recognise their specific ag using surface Ig
molecules

Antibodies

Dr Debasis Biswas

Ag recognition B cell stimulation.Blast

transformation and clonal proliferation
Effector Response Plasma cell formation
Secretion of specific antibodies
Time gap of 5-7 days..1ry response
Memory Response Memory B cell formation
2ry response more rapid, more intense

We already know

Expt. I: Tiselius & Kabat

Blood:

Plasma + Cells= Blood

Plasma:

Plasma - Fibrinogen= Serum

Rabbits immunized with egg albumin (OVA)......

Serum pooled and electrophoresed

Serum:

Blood allowed to stand ..

Clots..
Centrifuged at 3500 rpm X 10 mins
Supernatant is SERUM

Rabbits immunized with OVA.

Serum pooled; reacted in vitro with OVA......
Precipitate removed............................
Remaining serum electrophoresed

Serum

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globulin fraction contains antibodies

Immunoglobulins

Dimer of dimers
4 chain structure: L2H2

Drop in
globulin fraction

Serum

S-S bridges;
salt linkages; hydrogen
bonds; hydrophobic bonds
Light Chain

Heavy Chain

25000 Da

50000 Da

110 + 110 aa
(V + C)

110 + 330 aa/

110 + 440 aa
(V + C)

Glycoproteins:
CHOs increase solubility

Isotypes/ Antibody classes

Isotypes/ Antibody classes

Constant region of HC
5 basic patterns
..IgG
.IgA
.IgM
.IgD
.IgE

G1, G2,
G3, G4

Constant region of LC .
2 basic patterns

A1, A2

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Domains

X- ray Crystallographic Structure

Domain: 110 aa
IntrachainS-S:
60 aa loop

Hypervariable Regions/ CDRs

CDRs contact antigens

Variability= No. of aa at a sp. position/ Freq. of the commonest aa

at that position
Red. ..
Ag (Angiotensin II)
H1, H2, H3..
CDRs in Heavy Chain
L1, L2, L3..
CDRs in Light Chain
Yellow
Side Chains of CDRs
Contacting the Ag

Constant Region Domains

Hinge Region (IgG/ A/ D)
(rich in proline & cysteine)
flexibility to Fab fragments
CH2 Domain (IgG)
CH3 Domain (IgM)
Complement Activn.

Antibody- mediated Effector Functions

Complement Activation
Opsonization
ADCC (Antibody- depdt cell- mediated
cytotoxicity)
Transcytosis

CH3 Domain (IgG)

CH4 Domain (IgM)
Binding to Phagocytic cell.

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Complement Activation

Opsonization
Promotion of phagocytosis

C3b Receptors

Fc Receptors
bind C region of Ig

Bacteria

Bacteria

C3b
Antibodies
Antibodies
Immune complexes to liver/ spleen
RBC

Removal by resident macrophages

ADCC

Transcytosis

Fc Receptors
bind C region of Ig

Release of cytotoxic substances

like perforins & granzymes

Target cell
(virus- infected
cell)

Transport across epithelial layer..

Mucosal surfaces, viz. GI, Resp, GU, Breast
milk
Transport across placental barrier..
3rd trimester
IgA: major class
IgM: to mucosal surfaces
IgG: trans-placental

Antibodies

Expt. II: Behring & Kitasato

1st Nobel Prize in Medicine: Passive Immunization
Rabbits immunized with tetanus......
Serum pooled

6 control mice..
untreated

0.2 ml of serum injected into

abdominal cavity of 6 mice

24 hrs later
Live virulent tetanus bacilli injected to all mice
All treated mice..
HEALTHY

48 hrs later

All control mice..

DIED

Passive transfer of immunity through serum

Passive Antibody Therapy

For immunocompromised pts.

IVIG (Intravenous Immunoglobulin)
From pooled plasma of thousands of donors
Ab effector functions:
Complement Activn, Opsonization,
ADCC, Toxin Neutralisation

Risk of transmitting blood-borne pathogens

T/t with EtOH/ detergents

Eliminate Ab aggregates
Massive Complement Activn. ..Anaphylaxis

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IgA

IgG
Most abundant: 80% of total serum Ig
Most versatile:
Complement Activn: IgG3> IgG1> IgG2> IgG4
Opsonizn.:
IgG1> IgG3> IgG4> IgG2
ADCC:
Transcytosis:

1o- 15% of total serum Ig

2 mg/ ml
SecretionsBreast milk,
saliva, tears, mucosal
secretions
Monomeric. Serum
Dimeric/ Trimeric/
Tertameric.. Secretions

Class of ab with
highest daily prodn.

Secretory IgA

Secretory component: Protects hinge region from proteases

Bind to bacterial and viral surface antigens

Prevent their attachment to mucosa .
Inhibiting colonization & infection
Complexes of secretory IgA and antigen
entrapped in mucus and eliminated by ciliary
beating & peristalsis
Salmonella/ Vibrio cholerae/Polio/ Reovirus
Influenza virus
Neisseria gonorrhoeae

Breast milk: 1st month of life

IgM
5- 10% of total serum Ig
1.2 mg/ ml
Pentameric
Valency: 10
5
1ry immune response
1st ab in neonate
Complement Activn. (most
efficient class)
Diffuses poorly in
intercellular tissue fluids
Secreted in mucosal
secretions

IgD

0.2% of total serum Ig

30 g/ ml
Surface of B cells
Effector functions
unknown

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IgE

IgE- mediated . Mast- cell degranuln.

Build- up of cells for anti- parasitic defense

mIg + Ig- Ig= B- cell Receptor

Short cytoplasmic tail

Cannot interact with
signal transduction molecules