Adaptive immune system

Introduction

Adaptive Immune System

CMI Humoral Immunity

T-cells B-cells
Cytokines Antibodies
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 Cells of Adaptive Immunity

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Antigen recognition by Lymphocytes

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 Humoral immunity
Objectives
• Define what antibodies are

• Functions of antibodies

• Basic Structure of antibodies

• Classes of antibodies

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 Humoral immunity
• Mediated by antibodies
• Directed toward the defense against
– Extracellular microbes
– Toxins
• May culminate in the extracellular degradation
• Enhancement of their destruction via
phagocytosis

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 Antibodies /Immunoglobulins
• Soluble serum proteins secreted by activated B cells
(plasma cells) in response to an antigen
• Bind specifically with that antigen
• Antigen binding proteins of B-cell
– serve as B-cell receptors (IgM & IgD)
• Distributed in biologic fluids throughout the body
(Plasma, lymph, mucus, Tears etc.)
• Heterogeneous Abs are produced in response to
Multideterminant Ag
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 Monoclonal vs Polyclonal Antibodies
• Monoclonal Antibodies: Abs derived from a single
clone of B-cell and thus specific for a single epitope of
antigen

• Higher Specificity

• Polyclonal Antibodies: Abs derived from a variety of

B-cell clones that recognizes a particular epitope of
multivalent Ag

• Higher Sensitivity
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 Basic structure of antibodies
• All antibody molecules
share a common structure
of four peptide chains
• Two identical light chains
(Lc)
• Two identical heavy
chains (Hc)
• Chains linked together
with disulfide bonds
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 Ab stru…
• Each chain has a variable (V) region at one end and a
constant (C) region at the other end
• Variable regions of the heavy and light chains combine
to form the antigen-binding site
• The two arms of the Y end vary between different
antibody molecules- the variable (V) regions
• Antibodies responding to different antigens have
different V regions but the same C region in one class
(idiotype)
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 Ab stru…
Valency of antibody:
The number of antigenic determinants an individual
antibody can bind

• The valency of all antibodies is at least two

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 Ab stru…
• C regions form the stem of the Y-shaped antibody :

– Determine the class of the antibody (isotype)

– Serve common functions in all antibodies

– Dictate the cells and chemicals that the antibody

can bind to

– Determine how the antibody class will function in

elimination of antigens

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 Fab region
• Contained the antigen binding
sites of the antibody

• Contain both VH and VL

• Each antibody has different

antigen binding site

• Three segments of particular

variability in both the VH and
VL domains
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 Ab stru…
• They are designated hypervariable regions/
complementarity-determining regions

denoted as HV1, HV2, and HV3

• The most variable part of the domain is in the HV3

• Six HV regions in the Fab fragments (3 in the heavy

chain and 3 in the light chain)

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 Ab stru…
• Light chains contain 1 variable domain and 1
constant domain

• Heavy chains contain 1 variable domain and 3 or 4

constant domains

DOMAIN: a chain of about 60 aa, formed into a loop

by a single disulfide bond

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 Immunoglobulin classes
• Five different classes based on differences in the amino
acid sequences in the constant region of the heavy
chains (Isotype)

1. IgG -Gamma (γ) heavy chains

2. IgM -Micro (μ) heavy chains

3. IgA -Alpha (α) heavy chains

4. IgD -Delta (δ) heavy chains

5. IgE -Epsilon (ε) heavy chains

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 Immunoglobulin Subclasses
Based on Small differences in the amino acid sequences of
heavy chain constant regions
1. IgG Subclasses
a) IgG1 -Gamma 1 (γ1) heavy chains
b) IgG2 -Gamma 2 (γ2) heavy chains
c) IgG3 -Gamma 3 (γ3) heavy chains
d) IgG4 -Gamma 4 (γ4) heavy chains
2. IgA Subclasses
a) IgA1 -Alpha 1 (α1) heavy chains
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IgA2 -Alpha 2 (α2) heavy chains 16
 Immunoglobulin Types
• Based on Amino acid sequence in the constant
region of the light chain

1. Kappa light chains

2. Lambda light chains

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• Pentamer/monomer IgM
• Binds to ags with low affinity
• but it has high avidity-b/c of its
multimeric property
• Serve as : BCR

and complement activation

• The first Ig to be made in primary
infection
• Has short serum half-life : current
infection
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 IgG
• The major serum Ig -75% of serum Ig is IgG
• Produced late in the immune response
• It is monomeric and the smallest Ig in size
• Cross placenta and provide protection to fetus and
neonates: IgG2 does not cross
• It is a good opsonin
• Activate complement

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• About 19% of the Igs in normal IgA
serum

•Secreted as a dimer

•Most abundant in epithelial and

secretions (saliva, tears, GI tract,
Colostrum, sweat, and mucosal
lining)

•Plays role in mucosal immunity

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 IgE
• Lowest serum concentration <1%

• Secreted as monomeric

• Binds to mast cells, eosinophils and basophils

• Plays role in immunity against helminths

• It is also responsible for allergy (Reaginic antibody)

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 IgD
• Low concentration in serum (<1%)

• Monomer attached to the surface of B cells,

important in B cell activation

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 Function of Antibodies
• The functions of each Ab is attributed by both Fab
and Fc portion

A. Fab-mediated functions

1. Antigen recognition

• Abs bind specifically to all possible antigens

present in pathogens and their products

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 2. Neutralization
• Most bacteria and bacterial toxins are found outside the
cells

• Antibodies recognize antigens outside the cells

• Abs bind to pathogens and block its access to cells

• Prevent infection and destruction of host cells

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Neutralization

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Neutralization Abs: can block Viral infection of cells

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Antibodies can block adherence of bacteria to host
cells

E.g. Abs against pilin, the cell surface

protein of Neisseria gonorrhea which
enables it to adhere to the epithelial cells
of the urinary and reproductive tracts (&
essential to the bacterium’s infectivity),
inhibits adhesion & prevents infection

IgA: important to prevent adhesion of bacteria, viruses and

other pathogens 28
 Fc-mediated effector functions
• The Fc portion is responsible for several important
functions

• Activate accessory effector cells having surface Fc

receptors

– Phagocytic cells (macrophages and neutrophils)

– Natural killer cells, eosinophils and mast cells

• Enables them to identify and eliminate pathogens and

their undesirable products
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 Complement activation
• Antibody binding with pathogens leads to the formation
of Ag-Ab complex

• Ag-Ab complex activates the complement cascade

• Destruction and elimination of the invading pathogen

• IgG and IgM are potent complement activators

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Complement activation

Initiates the classical

complement cascade31
 Opsonization
• Antibodies Coat the surface of a pathogen

• Allow binding of their Fc domains to Fc receptors

present on effector cells

• Activation of phagocytosis and other killing

mechanisms

• Important to destruct and eliminate pathogens that

resist phagocytosis such as capsulated bacteria

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 The role of opsonization in immunity

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 Antibody-dependent cell-mediated
cytotoxicity (ADCC)
• Binding of Abs to pathogens

• Recruit NK and CT cells expressing Fc receptors

• Activation of NK cells

• Release of granules containing a membrane pore

forming protein that causes osmotic lysis

• A cytotoxin that activates DNA-degrading enzymes

• Destruction and clearance of infected cells

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 ADCC

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 Other uses for antibodies
• Diagnosis
• Therapeutics
• Vaccine

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 B-cell and T-cells interactions
• Most foreign antigens are thymus-dependent antigens
• B- cell response to such molecules requires the direct
contact of B cells with:
– TH cells and their cytokines
• B-cell endocytosed and processed foreign Ags after
regognizition
• Generate MHCII-Peptide complex
• Upregulated costimulatory molecule expression like
B7 on B-cell
• Present processed MHCII-Peptide to Th-cells , Th2
• Activate B-cells through cytokine and CD40L
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 B-cell –T-Cell….
• TH2 cytokines
• induce B-cell:
• Differentiation
• Memory
• Class switching
– Formation of
Different Abs
From IgM

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 B-cell –T-Cell….

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Phases of humoral immune response
for protein antigens
 Ab functions summary

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 Cytokines
Objectives
• Define Cytokines
• General Properties of Cytokines
• Which cells produce cytokines?
• Describe types and functions of cytokines
 Cytokines
• Soluble? Proteins that communicate among cells of the
immune system
• Secreted by many cells in response to microbes/others:
– Cells of immune system (innate and adaptive cells)
– Other cells: endothelial cells, epithelial cells
– Organs: liver, brain, endocrine glands
• Mediate & Regulate many of the functions of these cells
• Low molecular weight proteins,<30kD
• High affinity for receptors
– Active in picomole amounts
 Cytokines…
Interaction with its receptor on a target cell:
• Causes changes in the expression of adhesion
molecules on the target membrane
– Move from one location to another

• Signal an immune cell

– Increase/decrease the activity of particular
enzymes
– change its transcriptional program
– Altering and enhancing its effector functions
– Instruct a cell when to survive when to die
 Nomenclature of Cytokines
Cytokines have different names based on different
parameters:
• Source
– Lymphokines:-secreted by lymphocytes
– Monokines:-secreted by monocytes
• Function
– e.g. Chemokines: Mobilizing cells
• Intercellular action
– e.g. Interleukins
 Action and properties of Cytokines
 Cytokines exert their effect in different manners
• Autocrine
– Act on the generating
cell (self)
– Feature of most
cytokines
• Paracrine
– Act on cells in the
immediate vicinity
• Endocrine
– Affects cells remote
from the secreting cell
• Depends on their ability to
enter the circulation
& their half-life
Most cytokines have Autocrine and Paracrine
effect

T lymphocytes often secret cytokines at the site of

contact with antigen-presenting cells .
 Properties of Cytokines…
 Regulate the intensity and duration of the
immune response
– Stimulate or inhibit
activation/proliferation/differentiation
– Regulate the secretion of antibodies or other
cytokines
 Influence activity of B cells, T-cells, NK cells, Ms,
granulocytes, hematopoietic cells...
 Properties of Cytokines…
 Cytokine secretion is a brief, Self limited event:
– Cytokines are not stored as preformed molecules
 Cytokines often influence the Synthesis and actions of
other cytokine
– Cascading effect of one on the next
– Antagonize, Additive, Synergistic effects
 Cytokine actions may be local and systemic
– Bind to receptors on target cells
 Mediate and regulate the immune system
 Individual cytokines are produced by multiple cell types
 General Properties of Cytokines…
1. Pleiotropy: one cytokine has different effects on
different cells
2. Redundancy: two/more cytokines mediate the same
function
3. Synergism: the combined effect of two/more cytokines
is greater than the effect of the individual cytokines
4. Antagonism: when the effects of one cytokine inhibits
the effects of another cytokine
5. Cascade induction: the action of one cytokine on a
target cell induces that cell to produce other cytokines
that in turn induce other target cells
Cytokines can influence the expression of
cytokine receptors
 Functional categories of cytokines
• Different cytokines stimulate diverse responses of cells
• In the activation phase of adaptive immune responses
– stimulate the growth and differentiation of
lymphocytes
• In the effector phases of innate and adaptive immunity
– Activate different effector cells to eliminate microbes
• Stimulate the development of hematopoietic cells
• Therapeutic agents and targets for specific antagonists
in numerous immune and inflammatory diseases
 Functional….
Functionally cytokines can be categorized into 3 groups:
1. Mediators and regulators of innate immunity

Produced by macrophages, NK, monocytes…

▫ E.g. TNF, IL-1, IL-12, IFN-γ, Il-6, IL-18, Chemokines…

• Most act on endothelial cells and leukocytes to stimulate

the early inflammatory reactions to microbes

• Some function to control these responses

 2. Mediators and regulators of adaptive
immunity
IL-2, IL-4, IL-5, IFN-, TGF-…
– Produced by mainly T lymphocytes
– Some in the activation phase of T cell-dependent
immune responses
• Regulate growth and differentiation of various
lymphocyte populations
– Recruit, activate and regulate specialized effector cells
• Mononuclear phagocytes
• Neutrophils and eosinophils to eliminate antigens
in the effector phase of adaptive IR
Cytokines of the innate and adaptive immunity

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3. Stimulators of Hematopoiesis
• Stem cell factor (SCF), IL-7, IL-3…

• Produced by bone marrow stromal cells,

leukocytes, and other cells

• Stimulate the growth and differentiation of

immature leukocytes
Stimulators of Hematopoiesis
Functions of cytokines in host defense
Cytokines Belong to 4 Structural Families:

• Different amino acid sequences, similar structures

• Hematopoietin family (IL-2, IL-4)

• Interferon family (IFN-α, β, γ)
• Chemokine family
• Tumor necrosis factor (TNF) family
Receptor family and their ligands

 The receptors for most of the interleukins

belong to the class I cytokine receptor
family.
 C refers to conserved cysteine.
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 Cytokines that mediate and regulate innate
immunity
TNF
• Major source: Activated macrophages
• Ag-stimulated T cells, NK and mast cells can also secrete
• LPS major stimulant (from GM-Neg bacteria)
• IFN produced by T cells and NK cells augments TNF
synthesis by LPS-stimulated Ms
• Principal mediator of acute inflammatory response in
Gm-negative bacteria and other infectious microbes
– Recruit Neutro, Mono
 TNF…
• Causes vascular endothelial cells to express adhesion
molecules
• Stimulates endothelial cells and macrophages
– Secrete chemokines that enhance the affinity of
leukocyte integrins for their ligands and induce
leukocyte chemotaxis and recruitment
• Contributes to local inflammatory reactions that are
injurious to the host (e.g.in autoimmune diseases)
• Neutralizing Abs to TNF and soluble TNF receptors are
in clinical use to reduce inflammation in patients with
rheumatoid arthritis and inflammatory bowel disease
 TNF…
• Responsible for many of systemic complications of
severe infections
• TNF is produced in large amounts (enters blood stream)
• Acts on the hypothalamus to induce fever (IL-1 also)
• Acts on hepatocytes to increase synthesis of acute phase
proteins (also with IL-1 & IL=6)
• Large amount inhibits myocardial contractility and
vascular smooth muscle tone
– Marked fall in blood pressure, or shock
 TNF…
• TNF causes intravascular thrombosis
– Loss of the normal anticoagulant properties of the
endothelium (+ secrete activator of coagulation)
– Cause necrosis of tumors, which is the basis of its
name: thrombosis of tumor blood vessels
• High circulating levels of TNF cause severe metabolic
disturbances, such as a fall in blood glucose
concentrations to levels incompatible with life
– Due to overuse of glucose by muscle and failure of the
liver to replace the glucose
 TNF…
• TNF causes septic shock (endotoxin shock)
– A complication of severe gram-negative bacterial
sepsis

– Vascular collapse, disseminated intravascular

coagulation, and metabolic disturbances

– LPS-induced production of TNF and other

cytokines, including IL-12, IFN-y, and IL1
 IL-1
• Principal function similar to that of TNF
• Mediator of the host inflammatory response
• Functions with TNF
• Unlike TNF produced by other cells
– Neutrophils, endothelial cells, epithelial cells
• Similar biological function with TNF except
– IL-1 doesn’t induce apoptosis
• Does not cause the pathophysiologic changes of septic
shock
 IL-12
• Principal mediator of the early innate immune
response to intracellular microbes
• key inducer of CMI to these microbes
• Stimulate IFN- production by T cells & NK cells
• Main sources are Ms & DCs (activated Th cells
also induce production (CD40 on APCs-CD40L on T
cells interaction)
• Stimuli: LPS, intracellular bacteria (such as Listeria
& Mycobacteria), virus infections…
 IL-12
• Innate immunity against many microbes is mediated
by cytokines acting in the following sequence:
microbes  Ms response IL12  IFN- Ms
activation  killing of microbes

• Stimulates the differentiation of CD4 Th lymphocytes

into IFN- producing Th1 cells
 Type I Interferons (IFNs)
• Mediate innate immune response to virus

• Of two distinct groups of proteins: IFN- and IFN-

• Inhibits viral replication (Paracrine manner)

• Increases expression of class I MHC molecules

• Stimulates the development of TH1 cells in humans:

Expression of IL-12 receptor

• Increase the cytolytic activity of NK cells

 IL-10
• Inhibitor of activated Ms & DCs

– Inhibits the production of IL-12 by activated Ms

& DCs

– Inhibits the expression of costimulators and class

II MHC molecules on Ms & DCs

• Involved in the control of innate immune reactions

and CMI
Major Cytokines of innate immunity
 Cytokines Of Adaptive immunity
• Activation phase:

– Mediate proliferation and differentiation

• Effector phase:

– Mediate the activation of specialized effector

cells
 IL-2
• Growth factor for Ag-stimulated T lymphocytes

• Responsible for T cell clonal expansion after Ag

recognition (Autocrine)

• Promotes the proliferation and differentiation of other

immune cells

• Stimulates the growth of NK cells and enhances their

cytolytic function: lymphokine-activated killer cells

• Potentiates apoptotic death of Ag-activated T cells

 IL-4
• Development of TH2 cells from naive CD4 helper T cells
• The signature cytokine of the TH2 subset
– Inducer and an effector cytokine
• Stimulation of IgE and mast cell/eosinophil-mediated
reactions
• Suppression of M-dependent reactions
• Switching of IgM to the IgE isotype
( production of IgE Abs)
• Antagonizes the M-activating effects of IFN- and
thus inhibits cell-mediated immune reactions
 IL-5
• Activator of eosinophils
• produced by TH2 cells & activated mast cells
• Stimulate the growth and differentiation of
eosinophils
• Stimulates proliferation of B cells and production of
IgA Abs
• Eos express Fc receptors specific for IgE Abs
• IL-4 and IL-5 function in concert:
– IL4 stimulates production of IgE, which opsonizes
helminths and binds eosinophils and
– IL-5 activates the eosinophils to destroy the
parasites
 IFN- (type II IFN)
• The principal M -activating cytokine & serves critical
functions in innate + Adaptive immunity
• Some antiviral activity
• functions mainly as an effector cytokine of immune
responses
• The signature cytokine of TH1 cells
• NK cells secrete IFN-  in response to recognition of
microbes or in response to IL12
– Mediator of innate immunity
• T cells produce IFN-  in response to Ag recognition,
and enhanced by IL-12 and IL-18
 IFN-
• The sequence of reactions involving IL-12 and IFN- 
central to CMI against intracellular microbes.
• IFN- important in cell-mediated immunity against
intracellular microbes
– Macrophage-activating cytokine that provides the
means by which T lymphocytes and NK cells activate
macrophages to kill phagocytosed microbes
– IFN-  stimulates expression of class I and class II
MHC molecules and costimulators on PCs
• Potentiates many of the actions of TNF
• Differentiates naive CD4 T cells to the TH1 subset
• Inhibits the proliferation of TH2 cells
 IFN-
• Acts on B cells
– Promote switching to certain IgG subclasses,
– Inhibit switching to IL-4-dependent isotypes, such
as IgE
• Activates neutrophils and stimulates the cytolytic
activity of NK cells
 Transforming Growth Factor- (TGF- )
• Inhibits the proliferation and activation of lymphocytes
• Secreted by antigen-stimulated T cells, LPS-activated
mononuclear phagocytes…
• Some regulatory T cells produce TGF-
• Inhibits the proliferation and differentiation of T cells
and the activation of Ms
• Counteract the effects of proinflammatory cytokines
• Stimulates production of IgA antibodies
• Promote tissue repair
Biologic actions of IFN-
Major Cytokines of adaptive immunity
 Chemokines
• A subfamily of cytokines primarily functions
in directing migration of cells: chemotactic
cytokines or chemokines
• Chemoattractant Cytokines
 Chemokine Subclasses
• Most chemokines have 4 cysteine residues which
form disulphide bonds
• CC class: The first two cysteines are adjacent
(example: MCP-1, RANTES)
• CXC class: The first two cysteines are not adjacent
(example: IL-8)
• C class: Only has 2 cysteines not 4 (example:
Lymphotactin)
• CX3C class: Has 3 amino acids between the first two
cysteines and a different N-terminal
Chemokine Subclasses
Cytokine Network
 Major Histocompatibility Complex
(MHC) Molecule
Objectives
• Define MHC
• Discuss structure, function and properties of MHC
molecules
• Explain Peptide-MHC interaction
• Discuss expression of MHC molecules and
influencing factors
• Nature of antigen recognized by MHCI and MHCII
 MHC Molecule
• Large set of genes located on
chromosome 6

• Code for proteins (MHCI and

MHCII )

• Bind small peptides and present

them to T cells

• First detected by their effect on

transplant rejection – i.e tissue
Incompatibility
 MHC Genes
• MHC genome is called Human Leukocyte Antigen
(HLA) in human and H-2 in mice

• MHC genes are organized into 3 regions

• Class I MHC genes: HLA-A, HLA-B & HLA-C

• Class II MHC genes: HLA-DP, HLA-DQ, &HLA-DR

• Class III MHC genes: Complement factors C2, C4,&

Factor B
MHC Genes…
 MHC…
• All the MHC genes map within a single region of
the chromosome (hence called Complex)

• Genes encoding MHC III do not play any role in

antigen presentation
 Properties of MHC Genes
• MHC genes are:

– Polymorphic

Large number of alleles within each MHC class

– Polygenic: has several genes for each class

Vary considerably from person to person

– Co-dominantly expressed:

Each cell will express molecules inherited from both

parental alleles
MHC…
 MHC…
• Gene polymorphism leads to wide diversity of different
MHC molecules in an individual
• Diversity ensures that a wide range of peptides can be
presented to T-cells
• Different immune response will be evoked for Varity of
peptides
• Expression of MHC molecules is increased by cytokines
produced during innate and adaptive immune responses
 Expression of MHC molecule
• Class I expressed on all nucleated cells

• Class II expressed on APCs (Macrophages, B cells and

Dendritic cells)

• Expression of MHC molecules is increased by

cytokines produced during innate and adaptive
immune responses

 Tumor necrosis factor α and lymphotoxin (TNFβ)

 Interferon α, β and γ
Expression…
 Ag recognition by MHC molecule
• T-cells can only recognize MHC-peptide complex
displayed on other cells

• A peptide must associate with a given MHC of that

individual to induce immune response by T-cells

• The antigen binding sites of MHC molecule is flexible

• i.e The different peptides a given MHC molecule can

bind all bind to the same site, but only one at a time
 Ag recognition…
• MHC molecules only bind protein antigens:

• T-cells recognize and respond for protein Ags only

• Thus we can see that MHC molecules are antigen

presenting molecules
T Cells Only Recognize Antigen Associated with
MHC Molecules on Cell Surfaces
 T-cell-peptide–MHC complex
• MHC molecule binding site
on peptide antigen is
different from T-cell
binding site
 Class I MHC Molecule
• Presents peptide antigens to
CD8+ T-cells

• Can accommodate a peptide of

about 8-10 aminoacids (Close-
ended groove )
 Class I MHC…
• Has a single peptide binding cleft

• Can bind only one peptide at a time

• Presents peptides of endogenous proteins (from

cytosol): intracellular infections
 Structure of MHC I
• Made up of two polypeptide chains:

• One large α chain

• One small β2 microglobulin

• α1 and α2 domains form a cleft

where the Ag fragment binds

• α3 domain -site to which CD8 on T

cell binds

• The ends of the groove are closed

 Class II MHC Molecule
• Presents peptide antigens to
CD4+ T-cells

• Can accommodate a peptide of

about 13-18 aminoacids

• Open-ended groove
 Class II MHC
• Presents peptides of exogenous proteins (from
cytosol): extracellular infections

• Has a single peptide binding cleft

• Can bind only one peptide at a time

 Structure of MHC II
• Made up of two polypeptide
chains:

• One α chain and One β chain

• α1 and β1 domains form a cleft

where the Ag fragment binds

• β2 domain -site to which CD4 on

T cell binds

• The ends of the groove are open

MHC Summary
 Cell Mediated Immunity
Objectives
• Function of CMI
• How T cells recognise antigen?

• Sequence of Antigen processing

• Where does antigen processing take place?

• The bases of antigen processing mechanism

• Effector mechanisms of CMI

 Cell Mediated Immunity
• Mediated by T-Lymphocytes
• Helper T cells Ag specific
• Cytotoxic T cells
• Cytokines; IL-2 and IFN
• Macrophages...?(effector phase)
• Identify and eradicate antigens arise from inside the
cells of the body (Intracellular infections) expressed on
the surface of APCs bounded with MHC molecule
Defense against:
• Intracellular microbes inaccessible to antibodies:
Mycobacteria, Listeria monositogen
• Fungi, protozoa and helminths: Leishmania
• Cancer cells and Transplanted tissue
 T-Cells…
• The specificity of the response arises from T cells, but
the actual effector function is mediated by the
phagocytes
• T cells regulate proliferation and activity of other cells:
• B cells, macrophages, neutrophils…
– Shows the functional link between the adaptive
and innate immune responses
• Cell mediated immunity is not transferred to the fetus
• T-Cells have antigen receptor that recognizes and
reacts to a specific antigen
 Antigen recognition, processing and
presentation
• Unlike B cells, T cells cannot recognize
– Antigens in solution (Soluble antigens)
– Native antigens (not degraded)
– Antigens other than peptide antigens

• T-cells recognize peptide molecules presented with

MHC molecule on the surface of other cells (APCs)

• What is the importance of T-cell-MHC

restriction?
 Properties of Antigens Recognized by T -cells
• Most T lymphocytes recognize only peptides of protiens

• B cells can recognize peptides, proteins, nucleic acids,

polysaccharides, lipids

• T-cells recognize peptide molecules presented with

MHC molecule on the surface of cells

• Protein Ags should be degraded in to peptides so as to

form Peptide-MHC complexes

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T cells do not recognise native antigens

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 Antigens must be processed in order to be
recognised by T cells
T Y
Cell surface peptides of Ag
presented by cells that
express MHC antigens
Soluble
Soluble
native Ag
peptides
of Ag Cell surface
Cell surface
native Ag peptides
of Ag

ANTIGEN
PROCESSING
No T cell No T cell No T cell No T cell T cell
response response response response
response
 Antigen presenting cells (APCs)
• Immune cells that recognize, process and present
antigen to T-cells
• APCs express MHCII and Costimulatory molecules
• Display peptide-MHC complexes for recognition by T
cells
• Three professional APCs
– Dendritic cells
– Macrophages
– B-cells
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 APCs…

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T cells recognise MHC bound peptides

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 Antigen Processing
• Enzymatic process of
degrading proteins through
proteases into antigenic
peptides

• Occurs in proteasome

• APCs recognize, process and

present antigen to T-cells

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 Degradation in the proteasome
Cytoplasmic cellular proteins are degraded continuously
by a multicatalytic protease enzymes

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 Antigen presentation
• Transporting and displaying processed Peptides
associated with MHC molecule
• Peptide-MHC complexes takes place within the cell
• Cells that display MHC-associated peptides are called
antigen presenting cells (APCs)
• T cells from any one individual recognize foreign
peptide Ags only when they are bound to and displayed
by the MHC molecules of that individual
– Called self MHC restriction
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 Ag processing and presentation

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• CD4+ helper T cells recognize peptides bound to class II
MHC molecules (Class II MHC restricted)

• CD8+ T cells are class I MHC restricted

• CD4+ class II-restricted T cells recognize peptides

derived mainly from extracellular proteins that are
internalized into the vesicles of APCs: Exogenous Ags

• CD8 T cells recognize peptides derived from cytosolic,

usually endogenously synthesized, proteins

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 Steps of antigen processing and presentation
Uptake
• Access of native antigens and pathogens to
intracellular pathways of degradation
Degradation
• Limited proteolysis of antigens to peptides
Antigen-MHC complex formation
• Loading of peptides onto MHC molecules
Antigen Presentation
• Transport and expression of peptide-MHC
complexes on the surface of cells for recognition by
T cells
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 MHC molecule and Ag presentation

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T-cell subsets and
Effector mechanisms
 T Helper (TH) Cells
Central role in immune response
• Most are CD4+
• Recognize antigen on the surface of antigen
presenting cells (e.g macrophage) with MHCII
• Activate macrophages
• Induce formation of cytotoxic T cells
• Stimulate B cells to produce antibodies

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• Following activation by specific Ags presented by an
APC, Th cells initiate several key immune processes:

1) Selection of appropriate effector mechanisms ( e.g., B

cell activation or T cell generation);

2) Induction of proliferation of appropriate effector

cells and

3) Enhancement of the functional activities of other

cells (e.g., granulocytes, macrophages, NK cells)

131
 Activation of Th cells
• When naïve Th cells encounter Ag they activated and
differentiated into Th1, Th2 or other subsets
– Distinguished by the cytokines they produce
• Differentiation of Th0 cells depends upon the cytokines
in the environment
• Ags stimulate IL-4 production favors the generation of
Th2 cells
• Ags stimulate IL-12, favors the generation of Th1 cells
• IL 2 is important during proliferation of T cells
132
 Th1/Th2

Naïve Th0 IL-2, IL-4, IFN-g

-g

IL-12 IL-4

Effector
Effector
Th2 cell
Th1 cell IL-4, IL-6, IL-10
IL-2, IFN-g
 The T cell response involves activation,
proliferation and differentiation into effector cells

134
 Defferentiation of CD4+T cells

IL-22
IL-17A
Th22
IL-17F
Th17 IL-22
IL-21

IL-6; TNFα
IL-6; TGF-β IFN-γ
Th1
IL-12; IFN-γ
CD4+
T
IL-4 TGF-β
Activated DC Th2 Th9
IL-4
TGF-β IL-5
IL-13 IL-9
IL-12; IL-21

Bcl6 Treg
IL-4 IL-10
IL-21 Tfh TGF-β

135
Development of TH1 and TH2 subsets
 Effector functions of Th cells
• Th1: Produce IFN-γ, TNF B activate macrophages and
participate in the generation of cytotoxic lymphocytes
(CTL), resulting in a cell-mediated immune response
• IFN-γ suppresses the proliferation of Th2
• Th1 cells also associate with Delayed Type
Hypersensitivity

• Th2 cells: produce IL-4, IL-5, IL-13 that make Th2 cells
to be effective for immunity against helminthes
• IL-4 activates IgE production by B cells
• It suppresses differentiation into Th1
• IL-5 stimulates eosinophil, whereas IL-13 stimulate
mucus production
• Abnormal activity of Th2 cells result in allergy
138
TH1 – TH2 Crosstalk
Effector functions of Th1 cells
Effector functions of Th2 cells
 Effector mechanisms
employed by macrophages
include production of:
• TNF-α, which can induce
apoptosis

• Nitric oxide and other reactive

nitrogen intermediates

• Reactive oxygen intermediates

• Cationic proteins and

hydrolytic enzymes(lysozyme)
142
• Macrophage activation by Th1 cells
is very important in protection
against many different pathogens

• For example, Pneumocystis

carinii, an extracellular pathogen,
is controlled in normal individuals
by activated macrophages; it is,
however, a common cause of death
in AIDS patients because they are
deficient in Th1 cells.

• M. tuberculosis, an intracellular
pathogen that resides in vesicles, is
not efficiently killed by
macrophages unless they are
activated; hence this infection is a
problem in AIDS patients.
143
Development of TH1 vs. TH2 Response Determines
Clinical Outcome of Leprosy

Th1 Th2

144
Tuberculoid (L) vs. Lepromatous
leprosy (R)

Th1 Th2

145
 CD8+ T cells
• Cytotoxic T lymphocytes are activated by specific
antigen in the context of class I MHC, by Th1 cells
cytokines, especially IL-2, and IFN-γ

• CTL killing requires cell contact

• CTL are triggered to kill when they recognize the target
antigen associated with a cell surface MHC I molecule.
• CTLs are not injured when they lyse target cells
• Each CTL is capable of killing sequentially numerous
target cells

146
147
 Mechanisms of CTL-mediated killing
• CTLs express Fas ligand on their surface, which binds to
Fas receptors on target cells
• TNF-α bind to TNF receptors on target cells
• Fas and TNF receptors are a closely related familes
• Contain death domains that can activate caspases that
induce apoptosis in the target cell
• CTLs have numerous granules that contain Perforin
and Granzyme
• Perforin polymerizes to form channels in the target cell
membrane
• Granzymes, which are serine proteases, enter the target
cell through the channels and activate caspases and
nucleases in the target cell resulting in apoptosis
148
149
Mechanisms of CTL-mediated lysis
Summary

151
Summary...
 summary
• Two types of MHC molecules

MHC class I: Expressed by all nucleated cells

MHC class II: Expressed by APC

• Two types of T cells recognize MHC presentation

CD4 T helper cells

CD8 T cytotoxic cells

• Two types of antigens:

Endogenous (from within the cell)

Exogenous (from outside the cell)

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