CELLULAR MEDIATED

IMMUNE RESPONSE

Dr.Tetty Aman Nasution, MMedSc

Departemen Mikrobiologi FK USU
 Cell-Mediated Response
Two intracellular compartments:
• 1.Cytosolic: cytosol and nucleus connected
via nuclear pores
- site of viruses and some bacteria
• 2.Vesicular: membrane-bound entities
(endoplasmic reticulum, endosomes,
lysosomes, Golgi apparatus)
- site of some bacteria, some parasites
 Location of Pathogen Determines
Which T Cell Population Responds

• 1. Cytosolic: cytotoxic T cells (Tc) that

express CD8
• 2. Vesicular: subpopulation of helper T
cells (Th1) that express CD4
• 3. Extracellular: subpopulation of
helper T cells (Th2) that express CD4
 Cytotoxic (Tc) T Cells
A B

Cell expresses
viral antigens

Cytotoxic
C T cell

Virus infects cell

Infected cell is killed by cytotoxic T cell

by activation of nucleases that cleave
host and viral DNA
 Helper (Th1) T Cells
lysosome

Th1
Macrophage Macrophage cell

antigen
mycobacteria

Infected macrophage Activated infected

macrophage
Specificity of Immune Responses
Resides in Receptors
•T cell receptor (TCR) recognizes peptide
associated with major histocompatibility
complex (MHC) and is univalent.

•B cell receptor (surface immunoglobulin)

recognizes antigen and is bivalent

T B
cell cell
 Four Basic Principles of
Clonal Selection
1. Each lymphocyte bears a single type of
receptor of a unique specificity.
2. Interaction between a foreign
molecule and a lymphocyte receptor
capable of binding that molecule with
high affinity leads to lymphocyte
activation.
 Clonal Selection
3. Differentiated effector cells derived
from an activated lymphocyte will
bear receptors of identical specificity
to those of parental cell from which
the lymphocyte was derived.
4. Lymphocytes bearing receptors for
self molecules are deleted at an early
stage in lymphoid cell development.
Cells Expressing Class I and
Class II MHC

All nucleated cells

express class I
MHC
Class I MHC
Cells expressing
Class II class II MHC also
MHC
express class I
MHC
 Non-specific and
Specific Immunity
Specific (acquired, adaptive) immunity
• is induced and enhanced by antigen
• shows fine discrimination
• has memory

The non-specific and specific immune

systems interact with each other!
 Cells of the Immune System
• All derive from the bone marrow
• Two main lineages derive from the bone
marrow hematopoietic stem cells:
1. Lymphoid lineage
T cells, B cells, Natural Killer (NK) cells
2. Myeloid lineage
Monocytes, Macrophages, Dendritic cells,
Megakaryocytes, Granulocytes
 Hematopoiesis
Platelets Megakaryocyte

Tc
cell
Hematopoietic
Stem cell
Granulocyte TH
cell

Myeloid Lymphoid
progenitor progenitor
B cell
Mast cell

NK AFC
Dendritic cell
Plasma
Macrophage Monocyte cell
 Lymphocyte Recirculation
• Secondary lymphoid tissues (lymph
nodes, spleen) main sites where
lymphocytes encounter antigen
• Frequency of lymphocytes having a
receptor specific for a given antigen is
low
• Recirculation of lymphocytes through
lymphoid tissues optimizes productive
encounters with antigen to initiate
response
 Lymphocyte Recirculation
Naïve lymphocytes
enter lymph nodes
from the blood circulation
Lymphocytes return
to blood
via the thoracic duct

Antigens from infected area

go to lymph nodes
via the lymphatic system
 Leukocyte Migration and
Localization
• Bone marrow and thymus (primary
lymphoid tissues) produce B cells and T
cells, respectively
• B cells and T cells recirculate through
spleen and lymph nodes (secondary
lymphoid tissues)
• Antigen presenting cells (APC) pick up
antigen and migrate to secondary lymphoid
tissues and interact with T cells and B cells
 Leukocyte Migration and Localization
Bone marrow

Macrophage
Thymus
Dendritic
cell
T T B B
cell cell cell cell
Naive
T lymphocytes B
cell cell

Spleen and lymph nodes APC Tissues

Primed lymphocytes
T B
cell cell
Functions of Th Cells,
Th1 and Th2 Cells,
Macrophages, Tc Cells,
and NK Cells
 Critical Role of Th Cells in
Specific Immunity
• Select effector mechanisms
• Induce proliferation in appropriate effectors
• Enhance functional activities of effectors
Ag APC
Ag Antigen-presenting cell
Ag

Th B cell
cell

Cytokines
Tc
Granulocyte Macrophage NK
cell
NK cell
Cytokines
 Naïve Th Cells Can Differentiate
Into Th1 or Th2 Cells
Naive Th cells Short-term Chronic Long term
stimulation stimulation Memory cells
IFNγ
Th1 IL-2
cell
ThP ThO ThM
cell cell cell
Th2
cell

IL-2 IFNγ IL-2

IL-2 IL-4
IL-4 IL-5
IL-5 IL-6
IL-10 IL-10
Functions of Th1 and Th2 Cells
Th1 Th2
cell cell

Inhibits production
IL-10

IFNγ IL-4 IL-5

Activates Activates

Macrophage B cell Eosinophil

Mast cell

Antibodies (including IgE)

Central Role of Macrophages in
Natural and Specific Immunity
• Involved in initial defense and antigen
presentation and have effector functions
Invading agent Antigen presentation Activated macrophage

Macrophage Macrophage Macrophage

Th
cell

Cytokines
Cytokines Lymphokines Anti-microbial functions
Anti-tumor functions
 Detailed Functions of Macrophages
Inflammation – Fever, Production Damage to tissues
of: IL-6, TNF-alpha, IL-1 – act as Hydrolases, Hydrogen peroxide production
pyrogen Complement C3a
TNF alpha production
Immunity Antimicrobial action
Selection of lymphocytes to be
O2–dependent production of:
activated:
hydrogen peroxide, superoxide,
IL-12 results in Th1 activation
hydroxyl radical, hypochlorous acid
IL-4 results in Th2 activation
O2-independent production of:
Activation of lymphocytes:
acid hydrolases, cationic proteins,
Production of IL-1
lysozyme
Processing and presentation of
antigen Anti-tumor activity produced by:
Toxic factors
Reorganization of tissues, Hydrogen peroxide
Secretion of a variety of factors:
Complement C3a
Degradative enzymes (elastase,
Proteases, Arginase
hyaluronidase, collagenase)
Nitric oxide
Fibroblast stimulation factors
TNF alpha
Stimulation of angiogenesis
 Macrophage Activation
Macrophage activation results from
alterations in gene products that
govern new functions.
Two major mechanisms that activate
macrophages:
• IFN-γ produced by Th or Th1 cells plus
bacterial endotoxin (LPS)
• IFN-γ produced by Th or Th1 cells plus
TNF-α
Mechanism of Macrophage Activation
Macrophage 1 Activated
Macrophage
IFN gamma Bacterial endotoxin
TNF alpha
(lipopolysaccharide)
triggers cytokine
Th1 production
cell
IFN gamma

2 Activated Various
Macrophage Macrophage products
 Cytolytic T (Tc) Cells
• Tc exiting the thymus are pre-Tc cells,
i.e. have TCR that can recognize
antigen, but are not mature and cannot
kill until “armed”
• To become armed requires two signals:
1. Recognition by TCR of specific antigen
associated with class I MHC, and
2. Exposure to cytokines (IL-2 and IFN-γ)
 Mechanism of Arming Tc Cells
1. Cell expressing class I MHC
Class I presents antigen ( ) 2. Antigen-presenting
MHC to a pre-Tc cell cell presents antigen in
association with
3. Th cell class II MHC to Th cell
makes cytokines

Pre-Tc cell IFN T helper cell Class II MHC

IL-2 APC
4. Pre-Tc cell
differentiates to
functional Tc cell

Tc cell
6. Target cell
5. Tc recognizes antigen on is killed
class I MHC-expressing target cell
 Features of Tc Killing
• Antigen-specific
• Requires cell-cell contact
• Each Tc capable of killing many target
cells
 Main Mechanism of Tc Killing
• Tc granules contain perforin and granzymes
• Upon contact with target cell, granule
contents released, perforin polymerizes and
forms channel in target cell membrane
• Granzymes (serine proteases) enter target
cell through channel, activate caspases and
nucleases, lead to apoptosis of target cell
 Mechanism of Tc Killing

Tc cell Tc cell Granzymes

Perforin
monomers
Perforin Ca++
polymerizes

Polyperforin channels
Target cell Target cell
 Steps in Tc Killing
1. Tc recognizes antigen on
Target cell
Tc cell target cell

2. A lethal hit is delivered by

Tc cell Target cell the Tc using agents such as
perforin or granzyme B

3. The Tc detaches
Target cell from the target cell
Tc cell

Target cell 4. Target cell dies

by apoptosis
 Natural Killer (NK) Cells
• Derived from bone marrow
• Lack most markers for T and B cells (do not
have TCR)
• Do not undergo thymic maturation
• Express CD56, a specific NK marker
• Express a receptor for Fc portion of IgG,
called FcRIII (CD16)
• Cytokines (IL-2) promote differentiation
into lymphokine-activated killer (LAK) cells
 NK Cell Effector Mechanisms
• Mechanism of killing similar to those of Tc
cells
• Not MHC-restricted
• Susceptibility of target cell to killing is
inversely proportional to expression of
class I MHC (killer inhibitory receptors (KIR)
on NK cells recognize class I MHC and
prevent killing)
 NK Effector Mechanisms
(continued)
• IgG-coated target cells recognized by FcRIII
(CD16) are killed by antibody-dependent
cell-mediated cytotoxicity (ADCC)
• Lymphokine-activated killer cells (LAK) kill
broader range of cells than do NK cells
 Antigen

CD40
B T helper TCR
B MHC II cell
cell
cell
1. Antigen presentation to
Th cell B7 CD28
Immunoglobulin 2. B7 expressed
Cytokine 3. Th cell is
receptor receptor
activated
4. Cytokine
and expresses
binds to
CD40 ligand,
cytokine
Cytokines
receptor,
secreted
CD40 ligand
binds to CD40
CD40
ligand
B B B
cell cell cell B T helper
cell cell
5. B cell activated

6. B cells proliferate, differentiate, secrete Ig Cytokine

 Class II MHC B
Th cell APC Th cell
cell

Th cells are primed by antigen- B cell takes up and

presenting cell presents antigen

Th cell Th cell
B
cell B-T cell cooperation
B cells receive signals from T cells
B B
cell cell B cells divide

B B B B
cell cell cell cell

B
Antibody forming Antibody forming Antibody forming memory
cell cell cell cell
B Cells In Secondary Responses
• Memory cells created during primary
response
• Have high-affinity Ig receptors
• Can therefore take up antigens at
much lower concentrations than other
antigen presenting cells that lack Ig
antigen receptors
 Cytokines

Non-antibody proteins acting as

mediators between cells, termed:
• Monokines – mononuclear phagocytes
• Lymphokines – activated T cells,
especially helper T cells
• Interleukins – abbreviated IL with a
number
 Properties of Cytokines
1. Produced by cells involved in both
natural and specific immunity
2. Mediate and regulate immune
responses
3. Secretion brief and limited - not stored
pre-formed - synthesis initiated by
gene transcription - mRNA short-lived
- cytokines produced as needed
 Properties of Cytokines
(continued)

4. Can be produced by many cell types and act on

many cell types (pleiotropic)
5. Can have similar actions (redundant)

Receptor for: IL-2 IL-15 IL-7 IL-9 IL-4

IL15 Rα
IL2 Rα
IL2 Rβ

IL2 Rβ
IL2 Rγ

IL2 Rγ

IL2 Rγ

IL2 Rγ

IL2 Rγ
IL7 R

IL9 R

IL4 R
?
 Properties of Cytokines (continued)

6. Can influence synthesis of other cytokines

- produce cascades
- enhance or suppress production of other
cytokines
- exert positive or negative regulatory
mechanisms for immune responses
7. Influence action of other cytokines - can be
antagonistic, additive, synergistic
 Properties of Cytokines (continued)

8. Bind to receptors with high affinity

9. Cells responding to cytokine can be:
- same cell (autocrine)
- nearby cell (paracrine)
- distant cell by circulation (endocrine)
10. Cellular responses to cytokines are slow, require
new mRNA and protein synthesis
 Mediators and Regulators of
Natural Immunity
• Tumor Necrosis Factor-alpha (TNF-α)
• Interleukin-1 (IL-1)
• Chemokines (Chemotactic cytokines)
• Type I Interferons (IFN-α and IFN-β)
• Interleukin-12 (IL-12)
• Interleukin-10 (IL-10)
 Tumor Necrosis Factor (TNF-α)
• Produced by activated macrophages
• Most important mediator of acute
inflammation in response to microbes,
especially Gram-negative bacteria (LPS)
• Mediates recruitment of neutrophils and
macrophages to site of inflammation
• Acts on hypothalamus to produce fever
• Promotes production of acute phase
proteins
 Interleukin-1 (IL-1)

• Produced by activated macrophages

• Effects similar to those of TNF-α
 Chemokines

• Produced by many different leukocytes

and tissue cells
• Large family of >50 substances
• Recruit leukocytes to sites of infection
• Play a role in lymphocyte trafficking
 Type I Interferons (IFN-α and β)

• IFN-α a family of many proteins

produced by macrophages, IFN-β a
single protein produced by many cells
• Both IFNs inhibit viral replication
• Both increase expression of class I
MHC
• Both activate NK cells
 Interleukin-12 (IL-12)
• Produced by activated macrophages
and dendritic cells
• Stimulates production of IFN-γ
• Induces differentiation of Th cells to
become Th1 cells
• Enhances cytolytic functions of
cytotoxic T cells and NK cells
 Interleukin-10 (IL-10)
• Produced by activated macrophages,
Th2 cells
• An inhibitory cytokine
• Inhibits cytokine production by
activated macrophages
• Inhibits expression of class II MHC and
costimulatory molecules on
macrophages
 Mediators and Regulators of
Specific Immunity

• Interleukin-2 (IL-2)
• Interleukin-4 (IL-4)
• Interleukin-5 (IL-5)
• Interleukin-10 (IL-10)
• Interferon-gamma (IFN-γ)
 Interleukin-2 (IL-2)
• Produced by Th>>Tc
• Main growth factor for T cells

T cell
B cell Stimulation
of division
IL-2
Monocyte secretion
T cell
Stimulation
Activation
of division and IFN gamma
NK release (and other
Increase in NK
mediators)
Cell activity
 Autocrine Function of IL-2
Class II MHC
APC
Resting Activated
T cell T cell IL2 IL4 IL7

T cell Binds to IL-2 receptor

Division

T cell T cell T cell T cell

Receptor
decay

T cell T cell T cell T cell

 Interleukin-4 (IL-4)

• Produced by Th2 cells

• Stimulates Ig class switching to IgE
isotype
• Stimulates development of Th2 cells
from naïve Th cells
• Promotes growth of differentiated Th2
cells
 Interleukin-5 (IL-5)

• Produced by Th2 cells

• Promotes growth and differentiation
of eosinophils
• Activates mature eosinophils
• IL-4 and IL-5 can work together
Helminths opsonized with IgE can be
killed by activated eosinophils
 Interleukin-10 (IL-10)

• Produced by activated macrophages,

Th2 cells
• Inhibits production of IFN-γ by Th1
cells needed to activate macrophages
 Interferon-gamma (IFN-γ)
• Produced by Th cells >> Tc and NK cells
• Numerous functions in both natural and specific
immunity

NK Th1 cell >

Tc cell
Granulocyte
Endothelial cell
Activation
Activation IFN gamma
NK Increase in
Macrophage secretion NK
Activation cell activity

Many cell types Many cell types

B
T cell cell
Weak anti-viral activity,
Differentiation, Induction of class I
Stops cell division, T cell activation
Stops cell division and class II MHC
Stops hematopoiesis
 Cytokine Network

• Many cytokines have effects on many cells and

organ systems in addition to functions in immune
systems
• Referred to as the “cytokine network”
 Effects on Cells of Immune System

B NK LAK
cell T cell

Proliferation, Proliferation, Activation of cells of

Differentiation, Differentiation, immune system Cytokine production
Ig secretion Cytokine
and selection production

Macrophage
Lymphocyte cytokines
 Cytokine Effects on Organ Systems

Hypothalmus
Fever
Pituitary

ACTH
Lymphocyte Macrophage
cytokines
Adrenal
gland

Corticosteroid

Liver

Acute phase proteins

 Cytokine Effects on Various Cells
IL1 TNF IFNα
Fibroblasts IFNβ IFNγ
Fibrogenesis
Extracellular
matrix Lymphocyte Macrophage
cytokines
Many cell types
cytokines Anti-viral
state

IL1 TNF
Endothelial cell

Endothelial cells Tissue

Bone marrow
Fibrinogen remodeling
Hematopoiesis
Permeability change Tissue repair
adhesion Angiogenesis
Fibroblast
Extracellular
cytokines
matrix
Osteoclasts