Immunology and Immunotechnology

Q. Distinguish between:
I} HUMORAL AND CELL MEDIATED IMMUNITY
No
.

Attribute

Humoral

Cell mediated

Definition

The humoral response (or antibody

mediated response) involves B cells that
recognize antigens or pathogens that are
circulating in the lymph or blood
(humor is a medieval term for body
fluid).

The cellmediated response

involves mostly T cells and
responds to any cell that displays
aberrant MHC markers, including
cells invaded by pathogens, tumor
cells, or transplanted cells.

Antibodies

Mainly mediated by antibodies

Does not involve antibodies

Macromolecules
recognition

Can recognize protein, polysaccharide,

phospholipid and nucleic acid antigens

Can recognize only protein antigens

Antigens targetted

Immunity against:

Can act against soluble or free antigens

Provides immunity to extracellular

bacteria, viruses and toxins

Recognizes antigens presented by

APCs with Class I or Class II MHC
molecule
Provides immunity to intracellular
bacteria, viruses, fungi and
protozoa

II} INNATE AND ADAPTIVE IMMUNITY

No
.

Attribute

Components

Specificity

Response and
potency

Activity

Humoral
components

6
7

Innate
1.
2.
3.
4.
5.

Physical and chemical barriers

Phagocytic leukocytes
Dentritic cells
NK cells
Plasma proteins
Can recognise a broad range of
pathogens but unable to distinguish
small distinctions

Adaptive
1. Humoral immunity
2. Cell mediated immunity
Recognises highly specific
antigens

Immediate response and low potency

Slower response and high

potency

Always active

Normally shows silent action

Compliments
Acute phase reactants
Cytokines

Immunoglobulins

Diversity

Low. Hence limited specificity

Very high

Lag phase

Absent. Response is immediate

Present. Response takes time

Reacts with equal potency upon

repeated exposure to same antigen

Memory cells remember

specific antigens and reexposure of the same antigen
is faster than primary response

Inheritance

Inherited in genome

Not inherited

Antigen specificity

Not antigen specific

Antigen specific

Memory

9
10

III} B CELL RECEPTOR (IMMUNOGLOBULIN) AND T CELL RECEPTOR

Attributes

BCR

TCR

Structure

Transmembrane protein complex

composed of mIg

The TCR and chain each contains

two domains with the immunoglobulinfold structure

Composition

Has disulfide-linked heterodimers called

Ig/Ig

May have any of the two domain

structure or chains

Cytoplasmic
tail

Ig cytoplasmic tail contains 61 amino

acids and Ig has 48 amino acids

cytoplasmic tail contains 248 amino

acids and has 282 amino acids

Forms

Available in body in soluble form

T-cell receptor is membrane bound and

does not appear in a soluble form

Detection

Assessment of its structure can be done

by classic biochemical methods

Complex cellular assays are required for

assessment of its structure

No

Most T-cell receptors are specific not for

antigen alone but for antigen combined
with a molecule encoded by the major
histocompatibility complex (MHC).
Purification cannot be done by antigenbinding techniques, more advanced
methods required

Specificity

Specific for antigens alone

Purification

Purification can be done by simple

antigen-binding techniques

Distinguishing
regions

BCR has complementarity determining

regions (CDRs) in immunoglobulin light
and heavy chains

TCR variable domains have three

hypervariable regions

Constant region is directly connected to

transmembrane domain

In addition to the constant domain, each

TCR chain contains a short connecting
sequence, in which a cysteine residue
forms a disulfide link with the other
chain of the heterodimer.

Connection to
TM

10

Diagram

IV} ANTIGENS AND ANTIBODIES

No
.

Attribute

Antigens (immunogens)

Antibodies (immunoglobulins)

Definition

Any foreign substance that induces

immune response

Glycoproteins formed in the body

for combating antigens

Chemical Nature

Generally proteins, but can also be

carbohydrates, lipids or nucleic acids

They are glycoproteins

Characteristics

Triggers production of antibodes

Variable region has the antigen

binding domain

Binding region

Region on antigen that interacts with

antibodies is called Epitope

Variable region (CDRs) on

antibody that binds to epitope is
called Paratope

No. Of binding sites

An antigen may have many epitopes

Abs like IgG has only 2 paratope

Role

Cause disease or allergic reactions

Protect the system by

immobilizing or mediating lysis
of antigenic material

Structure

May be simple or highly complex

structure

Y shaped structure consisting of 2

heavy chains and 2 light chains

Source

May be endogenous or exogenous

Arise from plasma cells in

immune system

Types

10

Diagram

Many types present

5 classes IgG, IgM, IgE, IgD,

IgA

V} ENDOCYTIC AND CYTOSOLIC PATHWAY

No
.

Attribute

Endocytic pathway

Cytosolic pathway

Antigen involved

Exogenous antigens are processed

and presented by this pathway

Endogenous antigens are processed and

presented by this pathway

MHC

Class II MHC molecules

Class I MHC molecules

Antigen processing

Antigen processing happens in

Endocytic vesicles

Antigen processing happens in

Proteasome complex

Transport

The invariant chain guides transport

of class II MHC molecules
to endocytic vesicles

Transport of processed antigens to

endoplasmc reticulum is carried out by
TAP proteins

MHC association

Class II MHC and bind

invariant chain, blocking binding of
endogenous antigen.

Peptides assemble with class I MHC

aided by chaperone molecules
Calnexin, Caleticulin, Tapasin

Peptide-binding
groove

CLIP protein is present in peptidebinding groove as a block to avoid

any premature binding of antigenic
peptide

No blocking molecules present in

peptide-binding groove

Associated CD

CD4

CD8

Proteins that can be

degraded

Bacteria viruses, etc

Proteins with short half-lives;

denatured, misfolded, or otherwise
abnormal proteins

Summary of pathway

VI} DIRECT ELISA AND INDIRECT ELISA

No
.

Attribute

Direct ELISA

Indirect ELISA

Definition

It is the method wherein there is

only one set of antigens and one set
of antibodies to react.

It is a method in which 2 Antibodies

and a single antigen is involved

Sensitivity

Less sensitive

More sensitive

Antibody

Only one antibody involved which

is enzyme linked

Two antibodies involved primary and

enzyme linked secondary Ab

Immunoreactivity

Immunoreactivity of Primary
antibody may be affected because
of labelling

Immunoreactivity of Primary antibody

not affected.

Cross reactivity

Cross reactivity of secondary

antibody is eliminated

There are chances of cross reactivity

because of secondary antibody

Diagram

Flexibility

Different primary detection

antibodies have to be labelled
separately Less flexibility

Different primary detection antibodies

can be used with a single labelled
secondary antibody More flexibility

Methodology

Well coated with antigen is treated

with enzyme lined antibody and
later substrate for the enzyme is
added into the well to detect colour
change

Well coated with antigen is treated with

unlabelled primary antibody and later
enzyme linked secondary antibody is
added. The reaction is detected by
adding substrate for the enzyme

Reaction time

Time saving process

Time consuming process

10

Signal amplification

The signal is less amplified so this

is less sensitive method

The signal is more amplified so this is

more sensitive method