Cell mediated immunity

Historically, immunologists have divided adaptive immunity into;

1.Humoral immunity, which can be adoptively transferred from an immunized donor to a naive host by
antibodies in the absence of cells, and cell-mediated immunity, which can be adoptively transferred only
by viable T lymphocytes. The effector phase of humoral immunity is triggered by the recognition of
antigen by secreted antibodies; therefore, humoral immunity neutralizes and eliminates extracellular
microbes and toxins that are accessible to antibodies, but it is not effective against microbes inside cells.
2.Cell mediated immunity ,in the effector phase is initiated by the recognition of antigens by T cells. T
lymphocytes recognize protein antigens of microbes that are displayed on the surfaces of infected cells
as peptides bound to self major histocompatibility complex (MHC) molecules.

Introduction
Cell-mediated immunity is the type of host defense that is mediated by T lymphocytes, and it serves as
the defense mechanism against microbes that survive and replicate inside phagocytes and nonphagocytic
cells.
cell-mediated immunity is specific for cell associated microbes. Defects in cell-mediated immunity
result in increased susceptibility to infection by viruses and intracellular bacteria as well as some
extracellular bacteria and fungi that are ingested by phagocytes.
There are several important general principles of cell-mediated immune reactions.
Effector T cells of the CD4+ lineage link specific recognition of microbes with the recruitment and
activation of other leukocytes that destroy the microbes.
The nature of the leukocytes that are recruited and activated is determined by the subset of CD4+
effector T cells that are induced in the immune response.
TH1 cells activate macrophages, TH17 reactions are dominated by neutrophils (and variable numbers of
macrophages), and TH2 cells recruit and activate eosinophils.
Each type of leukocyte is specially adapted to destroy particular types of microbes.
T lymphocyte serves as a link between adaptive and innate immunity: by means of cytokine secretion
either by causing direct killing by inflammation or indirectly killing the cells by activating macrophages.
The adaptive immune response to microbes that are phagocytosed and live within the phagosomes of
macrophages is mediated by TH1 cells, which recognize microbial antigens and activate the phagocytes
to destroy the ingested microbes.
Many microbes have developed mechanisms that enable them to survive and even to replicate
within phagocytes, so innate immunity is unable to eradicate infections by such microbes. In these
situations, TH1 cells function to enhance the microbicidal actions of macrophages.
The response to extracellular microbes, including many fungi and bacteria, is mediated by TH17 cells.
These cells recruit neutrophils (and some monocytes).
The response to helminthic parasites is mediated by TH2 cells, which stimulate the production of
immunoglobulin E (IgE) antibodies and activate eosinophils and mast cells to eliminate the helminths.
The adaptive immune response to microbes that infect and replicate in the cytoplasm of various cell
types, including nonphagocytic cells, is mediated by CD8+ cytotoxic T lymphocytes (CTLs), which kill
infected cells and eliminate the reservoirs of infection.
CTL-mediated killing is also a mechanism for elimination of microbes that are taken up by phagocytes
but escape from phagosomes into the cytosol.
T cell–dependent inflammation may damage normal tissues.
Inflammation, consisting of leukocyte recruitmen and activation, accompanies many of the reactions of
CD4+ T lymphocytes and may be injurious under various conditions.
This T cell–dependent injurious reaction is called delayed-type hypersensitivity (DTH), the term
hypersensitivity referring to tissue damage caused by an immune response.
Cell-mediated immune responses consist of the development of effector T cells from naive cells in
peripheral lymphoid organs, migration of these effector T cells and other leukocytes to sites of infection,
and either cytokine mediated activation of leukocytes to destroy microbes or direct killing of infected
cells
MIGRATION OF EFFECTOR T LYMPHOCYTES TO SITES OF INFECTION
Some effector T cells exit the lymphoid organs where they were generated and preferentially home to
sites of infection in peripheral tissues, where they are needed to eliminate microbes during the effector
phase of adaptive immune responses.
The differentiation of naive T cells into effector cells, which occurs in the peripheral lymphoid organs, is
associated with a change in expression of the chemokine receptors and adhesion molecules that
determine the migratory behavior of these cells.
The expression of molecule involved in naive T cell homing into lymph nodes including L-selectin and
CCR7, decreases shortly after antigen-induced activation of the naive T cells, and the cell surface
expression of the sphingosine 1-phosphate receptor S1PR1 increases.
As a result, the effector cell that develop are no longer constrained to stay in the nod and are attracted to
enter the blood or efferent lymphatics which ultimately drain into the blood through the thoracic duct.
unlike naive T cells, effector T cells express chemokine receptors that bind chemokines produced at sites
of infection and adhesion molecules that bind to endothelial adhesion molecules that are induced on
postcapillary venules by cytokines, including IL-1 and tumor necrosis factor (TNF), that are produced at
infection sites.
TH1, TH2, and TH17 subsets of CD4+ T cells each have distinct homing phenotypes that direct them to
migrate into different sites of infections.
the chemokine receptors CXCR3 and CCR5, which bind to chemokine elaborated in tissues during
innate immune responses are expressed at high levels by TH1 cells but not by TH2cells.
TH1 cells tend to be abundant at sites of infection where the infectious agents trigger strong innat
immune reactions.
CTLs migrate in similar ways a TH1 cells.
In contrast, TH2 cells express the chemokine receptors CCR3, CCR4, and CCR8, which recognize
chemokine that are highly expressed at sites of helminth infection or allergic reactions.
TH17 cells express CCR6, which binds the chemokine CCL20. CCL20 is produced by various tissue
cells and macrophages in many bacterial and fungal infections.
The later, enhanced inflammation of effector T lymphocyte after exposure to pathogen in the infected
site is sometimes called immune inflammation.
The migration of effector T cells from the circulation to peripheral sites of infection is largely
independent of antigen, but cells that recognize antigen in extravascular tissues may be preferentially
retained there.
T cell migration through blood and lymphatic vessels is controlled mainly by adhesion molecules and
chemokines, which will engage T cells of any antigen specificity.
Once in the tissues, the T cells encounter microbial antigens presented by macrophages and other
antigen-presenting cells (APCs).
T cells that specifically recognize antigens receive signals through their antigen receptors that increase
the affinity of integrins for their ligands. Two of these integrins, VLA-4 and VLA-5, bind to fibronectin
in extracellular matrices, and a third adhesion molecule, CD44, which is also highly expressed on
effector and memory T cells, binds to hyaluronan.
As a result T cells not specific for the antigen that migrate into a site of inflammation may die in the
tissue or return through lymphatic vessels to the circulation.
(HELPER T CELL)
EFFECTOR FUNCTIONS OF CD4+ HELPER T CELLS
Effector T cells of the CD4+ lineage function by secreted cytokines and cell surface molecules to
activate other cells to eliminate microbes.
CD4+ T cells also participate indirectly in host defense by helping B lymphocytes to produce high-
affinity antibodies against extracellular microbes and by promoting the development of full functional
CTLs that combat intracellular microbes such as viruses.
The functions of CD4+ effector cells in cell-mediated immunity can be divided into several steps ;
1.Recruitment of other leukocytes - The recruitment of neutrophils, monocytes, and eosinophils to the
site of the reaction is mediated by chemokines produced by the T cells themselves and by other cells in
response to cytokines secreted by the T cells.
Activation of the recruited leukocytes - The mechanisms by which CD4+ T cells activate other
leukocytes involve T cell expression of the surface protein CD40 ligand (CD40L) and secretion of
cytokines.
The CD40Lmediated pathway is best defined for TH1-mediated activation of macrophages.
Amplification of the response - For instance, cytokines produced by T cells activate macrophages to
produce other cytokines that in turn act on the T cells and increase their responses.
 Down regulation of the response - Special control mechanisms may also operate to limit effector
responses after the antigen is eliminated. For instance, both TH1 cells and activated macrophages
produce the cytokine IL-10, which functions mainly to inhibit further TH1 differentiation and
macrophage activation.
Additional inhibitory mechanisms, such as other anti-inflammatory cytokines and receptors that turn off
T cell activation, may also be involved in controlling T cell–mediated responses.
Functions of TH1 Cells
The principal function of TH1 cells is to activate macrophages to ingest and destroy microbes
The same reaction of TH1-mediated macrophage activation is involved in injurious delayed-type
hypersensitivity, which is a component of many inflammatory diseases, and in granulomatous
inflammation, which is typical of tuberculosis and is also seen in some other infectious and
inflammatory disorders.
Cytokines Produced by TH1 Cells
The signature cytokine of TH1 cells is IFN-γ. TH1 cells also produce TNF, some chemokines, and other
cytokines.
Interferon-γ, IFN-γ is the principal macrophage-activating cytokine and serves critical functions in
immunity against intracellular
microbes.
IFN-γ is a homodimeric protein belonging to the type II cytokine family. In addition to CD4+ TH1 cells,
IFN-γ is also produced by NK cells and CD8+ T cells.
The receptor for IFN-γ is composed of two structurally homologous polypeptides belonging to the type
II cytokine receptor family, called IFNγR1 and IFNγR2.
IFN-γ binds to and induces the dimerization of the two receptor chains. This leads to activation of the
associated JAK1 and JAK2 kinases and ultimately to phosphorylation and dimerization of STAT1,
which stimulates transcription of several gene.
IFN-γ–induced genes encode many different molecules that mediate the biologic activities of this
cytokine.
The function includes;
IFN-γ activates macrophages to kill phagocytosed microbes, the hallmark of “classically activated”
macrophages.
In adaptive cell-mediated immunity, IFN-γ produced by TH1 cells works together with CD40 ligand,
also expressed by the T cells, to activate macrophages.
IFN-γ activates numerous signaling pathway, most importantly STAT1, and TLR and CD40 signals
activate the transcription factors nuclear factor κB (NF-κB) and activation protein 1 (AP-1).
These transcription factors stimulate the expression of several enzymes in the phagolysosomes of
macrophages, including phagocyte oxidase, which induces the production of reactive oxygen species
(ROS); inducible nitric oxide synthase (iNOS), which stimulates the production of nitric oxide (NO);
and lysosomal enzymes. These substances destroy ingested microbes in the vesicles.
IFN-γ acts on B cells to promote switching to certain IgG subclasses, notably IgG2a or IgG2c) (in mice),
an to inhibit switching to IL-4–dependent isotypes, such as IgE.
The IgG subclasses induced by IFN-γ bind to Fcγ receptors on phagocytes and activate complement, and
promote the phagocytosis of opsonized microbes.
IFN-γ promotes the differentiation of CD4+ T cells to the TH1 subset and inhibits the differentiation of
TH2 and TH17 cells. These actions of IFN-γ serve to amplify the TH1 response.
IFN-γ stimulates expression of several different proteins that contribute to enhanced MHC-associated
antigen presentation and the initiation and amplification of T cell–dependent immune responses.
These proteins include MHC molecules; many proteins involved in antigen processing, including the
transporter associated with antigen processing (TAP); components of the proteasome; HLA-DM; and B7
co stimulators on APCs.
Other TH1 Cytokines
In addition to IFN-γ, TH1 cells produce TNF and various chemokines, which contribute to the
recruitment of leukocytes and enhanced inflammation.
TH1 cells are also important sources of IL-10, which functions mainly to inhibit dendritic cells and
macrophages and thus to suppress TH1 activation (Negetive feedback).
TH1-Mediated Classical Macrophage Activation and Killing of Phagocytosed Microbes
At any site of infection, as part of the innate immune response, monocytes are recruited from blood into
tissues
by chemokines produced by macrophages and other cells resident at the site.
These monocytes mature into tissue macrophages and first attempt to phagocytose and destroy the
pathogen.
If the microbe survives within the the phagosomes. In these infected cells, microbial peptides are
processed and presented as peptides associated with class II MHC molecules.
These T cells are recruited to the site of infection, where they recognize antigenic peptides.
The macrophages are exposed to signals from the TH1 effector cells, which activate the macrophages to
kill the ingested microbes.
CD4+ TH1 cells activate macrophages by contact mediated signals delivered by CD40L-CD40
interactions and by IFN-γ. When the TH1 cells are stimulated by antigen, the cells express CD40L on
their surface and secrete IFN-γ. The actions of IFN-γ on macrophages, described earlier, synergize with
the actions of CD40 ligand (on macrophage surface), and together they are potent stimuli for
macrophage activation.
T cell–dependent activation of B lymphocytes—helper T cells stimulate B lymphocyte proliferation and
differentiation by CD40-mediated signals and cytokines.
Activated macrophages kill phagocytosed microbes mainly by the actions of reactive oxygen species,
nitric oxide, and lysosomal enzymes. All these potent microbicidal agents are produced within the
lysosomes of macrophages and kill ingested microbes after phagosomes fuse with lysosomes. These
toxic substances may also be released into adjacent tissues, where they kill extracellular microbes and
may cause damage to normal tissues. This pathway of macrophage activation is called classical
activation.
Activated macrophages stimulate inflammation through the secretion of cytokines, mainly TNF, IL-1,
and chemokines, and short-lived lipid mediators such as prostaglandins, leukotrienes, and
plateletactivatingfactor.
The collective action of these macrophage-derived cytokines and lipid mediators is to recruit more
leukocytes.
Activated macrophages amplify cell-mediated immune responses by becoming more efficient APCs
because of increased levels of molecules involved in antigen processing and increased surface
expression of class II MHC molecules and co stimulators and by producing cytokines (such as IL-12)
that stimulate T lymphocyte differentiation into effector cells.
The microbicidal products released by activated macrophages and neutrophils are capable of injuring
normal tissue and do not discriminate between microbes and host tissue. However the extent of injury is
less.
Functions of TH2 Cells
TH2 cells stimulate IgE- and eosinophil-mediated reactions that serve to eradicate helminthic infections.
Helminths are too large and may be too resistant to be phagocytosed by the neutrophil Therefore, TH2
cells secrete IL-4, IL-5, and IL-13, which work cooperatively to eradicate these infections.
Cytokines Produced by TH2 Cells
The functions of TH2 cells are mediated by IL-4, which induces IgE antibody responses; IL-5, which
activates eosinophils; and IL-13, which has diverse actions.
Interleukin-4
IL-4 is the major stimulus for the production of IgE antibodies and for the development of TH2 cells
from naïve CD4+ helper T cells.
It functions as both an inducer and an effector cytokine of these cells. IL-4 is a member of the four–α-
helical cytokine family .
The IL-4 receptor of lymphoid cells consists of a cytokine-binding α chain that is a member of the type I
cytokine receptor
family, associated with the γc chain shared by other cytokine receptors. This IL-4Rαγc receptor signals
by the JAKSTAT pathway (JAK3 or JAK4 and STAT6) and by a pathway that involves the insulin
response substrate (IRS) called IRS-2. IL-4 and IL-13 activate the STAT6 protein, which induces
transcription of genes that account for many of the actions of these cytokines.
IL-4 also bindsto the IL-13 receptor .
IL-4 has important actions ;
1.IL-4 stimulates B cell Ig heavy chain class switching to the IgE isotype. IgE antibodies play a role in
eosinophil-mediated defense against helminthic (and some arthropod) infections.
IgE is also the principal mediator of immediate hypersensitivity (allergic) reactions .
IL-4 also enhances switching to IgG4 (in humans, or the homologous IgG1 in mice) and inhibits
switching to the IgG2a and IgG3 isotypes in mice, both of which are stimulated by IFN-γ.
2. IL-4 stimulates the development of TH2 cells and functions as an autocrine growth factor for
differentiated TH2 cells.
3. IL-4, together with IL-13, contributes to an alternative form of macrophage activation. IL-4 and IL-13
suppress IFN-γ–mediated classical macrophage activation and thus inhibit defense against intracellular
microbes.
4. IL-4 (and IL-13) stimulate peristalsis in the gastrointestinal tract, and IL-13 increases mucus secretion
from airway and gut epithelial cells. Both these actions contribute to elimination of microbes at
epithelial surfaces.
5. IL-4 and IL-13 stimulate the recruitment of leukocytes, notably eosinophils, by promoting expression
of adhesion molecules on endothelium and the secretion of chemokines that bind chemokine receptors
expressed on eosinophil.
Interleukin -13
IL-13 is a member of the four–α-helical cytokine family, with limited sequence homology but
significant structural similarity to IL-4 .
The functional IL-13 receptor is a heterodimer of the IL-4Rα chain and the IL-13Rα1 chain. This
complex can bind both IL-4 and IL-13 with high affinity.
The receptor is expressed on B cells, mononuclear phagocytes, dendritic cells, eosinophils, basophils,
fibroblasts, endothelial cells, and bronchial epithelial cells.
Some of the actions of IL-13 overlap those of IL-4, and others are distinct. IL-13 functions with IL-4 to
induce alternative macrophage activation, which contributes to tissue repair and fibrosis. IL-13
stimulates mucus production by airway epithelial cells, an important component of allergic reactions
such as asthma.
Unlike IL-4, IL-13 is not involved in TH2 differentiation.
Interleukin-5
IL-5 is an activator of eosinophils and serves as the principal link between T cell activation and
eosinophilic inflammation. It is a homodimer of a polypeptide containing a four–α-helical domain and is
a member of the type I cytokine family.
The IL-5 receptor is a heterodimer composed of a unique α chain and a common β chain (βc), which is
also part of the IL-3 an granulocytemacrophage colony-stimulating factor (GM-CSF) receptors .
The major IL-5–induced signaling pathway involves JAK2 and STAT3.
The principal actions of IL-5 are to activate mature eosinophils and to stimulate the growth and
differentiation of eosinophils. Eosinophils express Fc receptors specific for IgE and some IgG antibodies
and are thereby able to bind to microbes, such as helminths.
Roles of TH2 Cells in Host Defense
1. IgE- and eosinophil-mediated reactions - IL-4 (and IL-13) stimulates the production of helminth-
specific IgE antibodies. IL-5 activates the eosinophils and these cells release their granule contents,
including major basic protein and major cationic protein, which are capable of destroying even the tough
integuments of helminthes.
2. Activation of mast cells- Mast cells express high affinity Fcε receptors and may be activated by IgE
coated Helminthes. mast cells secrete cytokines such as TNF and chemokines, and lipid mediators, all of
which induce local inflammation that helps to destroy the parasites.
3. Barrier immunity- Cytokines produced by TH2 cells are involved in blocking entry and promoting
expulsion of microbes from mucosal organs. e.g. IL-13 stimulates mucus production, and IL-4 and IL-13
may stimulate peristalsis in the gastrointestinal system. Thus, TH2 cells play an important role in host
defense at the barriers.
4. Alternative macrophage activation- IL-4 and IL-13 activate macrophages to express enzymes that
promote collagen synthesis and fibrosis. The macrophage response to TH2 cytokines has been called
alternative macrophage activation to distinguish it from the activation induced by IFN-γ (Classical
immunity).
Alternatively activated macrophages may also serve to initiate repair after diverse types of tissue injury.
Alternatively activated macrophages induce the formation of fibrous tissue by secreting growth factors
that stimulate fibroblast proliferation (platelet-derived growth factor), collagen synthesis (transforming
growth factor-β [TGF-β]), and new blood vessel formation or angiogenesis (fibroblast growth factor).
TH2 cytokines suppress classical macrophage Activation and interfere with protective TH1- mediated
immune responses. IL-4 stimulates production of cytokines such as IL-10 and TGF-β that inhibit TH1
development and function.
Functions of TH17 Cells
TH17 cells secrete cytokines that recruit leukocytes, mainly neutrophils, to sites of infection.
Cytokines Produced by TH17 Cells
Interleukin-17
The IL-17 family includes six structurally related proteins, of which IL-17A and IL-17F are the most
similar and are produced by TH17 cells.
IL-17 receptors are multimeric and expressed on a wide range of cells.
The function includes;
1. IL-17 induces neutrophil-rich inflammatory reactions. It also enhances neutrophil generation by
increasing the production of G-CSF and the expression of its receptors.
2. IL-17 stimulates the production of antimicrobial substances, including defensins, from numerous cell
types.
Other TH17 Cytokines
1.IL-22 is a member of the IL-10 cytokine family. It is produced particularly TH17 cells, and by NK
cells. it is produced in epithelial tissues, especially of the skin and gastrointestinal tract, and serves to
maintain epithelial integrity, mainly by promoting the
barrier function of epithelia and by stimulating repair reactions.
2. IL-21 is produced by activated CD4+ T cells, including TH17 cells. The IL-21 receptor belongs to the
type I cytokine receptor family, consists of a ligand binding chain and the γc subunit, and activates a
JAKSTAT signaling pathway.
IL-21 is required for the generation of follicular helper T cells and is also produced by follicular helper
cells and stimulates B cells in germinal centers. IL-21 has also been shown to promote the
differentiation of TH17 cells, especially in humans, providing an autocrine pathway for amplifying
TH17 responses.
Roles of TH17 Cells in Host Defense
1.The ability of IL-17 to recruit neutrophils accounts for the central role of TH17 cells in adaptive
immune reactions in which neutrophilic inflammation is prominent. The recruited neutrophils ingest and
kill extracellular microbes, including fungi and bacteria.
2. TH17 cells are also important in the pathogenesis of to many inflammatory diseases, such as
psoriasis, inflammatory bowel disease, rheumatoid arthritis, and multiple sclerosis.
(cytotoxic T cell)
EFFECTOR FUNCTIONS OF CD8+ CYTOTOXIC T LYMPHOCYTES
CD8+ CTLs eliminate intracellular microbes mainly by killing infected cells.
The development of a CD8+ CTL response to infection proceeds through antigen-mediated stimulation
of naive CD8+ T cells in lymphoid organs, clonal expansion, differentiation, and migration of
differentiated CTLs into tissues.
In addition to direct cell killing, CD8+ T cells secrete IFN-γ and thus contribute to macrophage
activation in host defense and in hypersensitivity reactions.
CTL-mediated killing involves specific recognition of target cells and delivery of proteins that induce
cell death.
CTLs kill targets that express the same class I–associated antigen that triggered the proliferation and
differentiation of naive CD8+ T cells from which they are derived and do not kill adjacent uninfected
cells that do not express this antigen.
The killing is highly specific because an “immunologic synapse” is formed at the site of contact of the
CTL and the antigen-expressing target, and the molecules that actually perform the killing are secreted
into the synapse and cannot diffuse to other nearby cells.
The process of CTL-mediated killing of targets consists of antigen recognition, activation of the CTLs,
delivery of the “lethal hit” that kills the target cells, and release of the CTLs .

Recognition of Antigen and Activation of CTLs

The CTL binds and reacts to the target cell by using its antigen receptor, coreceptor (CD8), and adhesion
molecules.
To be efficiently recognized by CTLs, target cells must express class I MHC molecules complexed to a
peptide (the complex serving as the ligand for the T cel receptor (TCR) and the CD8 coreceptor) and
intercellular adhesion molecule 1 (ICAM-1, the principal ligand for the LFA-1 integrin).
This immunologic synapse formed between the two cells is characterized by a ring of close apposition
between the CTL and target cell membranes, mediated by LFA-1–ICAM-1 binding, and an enclosed gap
or space inside the ring.
This interaction results in the initiation of biochemical signals that activate the CTL, which are
essentially the same as the signals involved in the activation of helper T cells
Cytokines and costimulators provided by dendritic cells, which are required for the differentiation of
naive CD8+ T cells into CTLs, are not necessary for triggering the effector function of CTLs (i.e., target
cell killing).
Therefore, once CD8+ T cells specific for an antigen have differentiated into fully functional CTLs, they
can kill all nucleated cell that shows the same antigen.
In addition to the T cell receptor, CD8+ CTLs express receptors that are also expressed by NK cells.
Some CTLs.
Some of these receptors belong to the killer immunoglobulin receptor (KIR) family that recognizes
class1 MHC molecule. These KIRs transduce inhibitory signals that may serve to prevent CTLs from
killing normal cells.
CTLs express the NKG2D receptor, that recognizes class I MHC–like molecules MIC-A, MIC-B, and
ULBP, expressed on infected or neoplastic cells.
Killing of Target Cells by CTLs
Within a few minutes of a CTL’s antigen receptor recognizing its antigen on a target cell, the target cell
undergoes changes that induce it to die by apoptosis.
Target cell death occurs during the next 2 to 6 hours and proceeds even if the CTL detaches. Thus, the
CTL is said to deliver a lethal hit to the target cell.
The principal mechanism of CTL-mediated target cell killing is the delivery of cytotoxic proteins stored
within cytoplasmic granules (also called secretory lysosomes) to the target cell.
CTL recognition of the target leads to activation of the CTL, one consequence of which is cytoskeleton
reorganization such that the microtubule organizing center of the CTL moves to the area of the
cytoplasm near the contact with the target cell.
The cytoplasmic granules of the CTL are transported along microtubules and become concentrated in
the region of the synapse, and the granule membrane fuses with the plasma membrane at the secretory
domain.
Membrane fusion results in exocytosis of the CTL’s granule contents into the confined space within the
synaptic ring, between the plasma membranes of the CTL and target cell.
The cytotoxic proteins in the granules of CTLs (and NK cells) include granzymes and perforin.
Granzymes A, B, and C are serine proteases that cleave proteins after aspartate residues. Perforin is a
membrane-perturbing molecule homologous to the C9 complement protein.
The granules also contain and a sulfated proteoglycan, serglycin, which serves to assemble a complex
containing granzymes and perforin.
The main function of perforin is to facilitate delivery of the granzymes into the cytosol of the target cell.
i.Perforin may polymerize and form aqueous pores in the target cell membrane through which
granzymes enter
ii.According to another current model, complexes of granzyme B, perforin, and serglycin are discharged
from the CTL onto the target cell and are internalized into endosomes by receptor-mediated endocytosis.
Perforin may act on the endosomal membrane to facilitate the release of the granzymes into the target
cell cytoplasm. Once in the cytoplasm, the granzymes
cleave various substrates, including caspases, and initiate apoptotic death of the cell.
Another protein found in human CTL (and NK cell) granules, called granulysin, can alter the
permeability of target cell and microbial membranes.
CTLs also use a granule-independent mechanism of killing, On activation, CTLs express a membrane
protein, called Fas ligand (FasL), that binds to the death receptor Fas, which is expressed on many cell
types. This interaction also results in activation of caspases and apoptosis of Fas-expressing targets.
After delivering the lethal hit, the CTL is released from its target cell, which usually occurs even before
the target cell goes on to die.
In addition, CTL granules contain a proteolytic enzyme called cathepsin B, which is delivered to the
CTL surface on granule exocytosis, where it degrades errant perforin molecules that come into the
vicinity of the CTL membrane.
Roles of CD8+ CTLs in Host Defense
There are two types of situations in which cells cannot destroy microbes that infect them. First, some
viruses live and replicate in cells that are incapable of destroying microbes (such as hepatitis viruses in
liver cells). Second, even in phagocytes, some microbes escape from vesicles and live in the cytoplasm,
where the microbicidal mechanisms of phagocytes are ineffective.
Such infections can be eliminated only by destroying the infected cells, and in adaptive immune
responses, CD8+ CTLs are the principal mechanism for killing infected cells.
By activating nucleases in target cells, CTLs can initiate the destruction of microbial DNA as well as the
target cell genome, thereby eliminating potentially infectious DNA.
The massive expansion of CD8+ T cells that follows infections provides a large pool of CTLs to combat
these infections.
Destruction of infected cells by CTLs is a cause of tissue injury in some diseases. For instance, in
infection by hepatitis B and C viruses, the infected liver cells are killed by the host CTL (and NK cell)
response and not by the viruses.

Suppressor T cell
It is a type of immune cell that blocks action of other type of lymphocytes ,to keep the immune system
from becoming overactive.
It is also called regulatory T cell ,T reg and T regulatory cell.
Tregs are immunosuppressive and generally suppress or downregulate induction and proliferation of
effector T cells.
Tregs express the biomarkers CD4, FOXP3, and CD25 and are thought to be derived from the same
lineage as naïve CD4 cells.
Because effector T cells also express CD4 and CD25, Tregs are very difficult to effectively discern from
effector CD4+.cytokine TGFβ is essential for Tregs to differentiate from naïve CD4+ cells and is
important in maintaining Treg homeostasis.
Mouse models have suggested that modulation of Tregs can treat autoimmune disease and cancer and
can facilitate organ transplantation and wound healing.
Tregs tend to be upregulated in individuals with cancer, and they seem to be recruited to the site of many
tumors.
High numbers of Tregs in the tumor microenvironment is indicative of a poor prognosis, and Tregs are
thought to suppress tumor immunity, thus hindering the body's innate ability to control the growth of
cancerous cells.
This is an important "self-check" built into the immune system to prevent excessive reactions.
Regulatory T cells come in many forms with the most well-understood being those that express CD4,
CD25, and FOXP3 (CD4+CD25+ regulatory T cells). Another regulatory T cell subset is Treg17 cells.
Regulatory T cells are involved in shutting down immune responses after they have successfully
eliminated invading organisms, and also in preventing autoimmunity.
Additional regulatory T cell populations include Tr1, Th3, CD8+CD28-, and Qa-1 restricted T cells.
Development
All T cells begin as CD4-CD8-TCR- cells at the DN (double-negative) stage, where an individual cell
will rearrange its T cell receptor genes to form a unique, functional molecule, which they, in turn, test
against cells in the thymic cortex for a minimal level of interaction with self-MHC. If they receive these
signals, they proliferate and express both CD4 and CD8, becoming double-positive cells.
The selection of Tregs occurs on radio-resistant hematopoietically-derived MHC class II-expressing
cells in the medulla or Hassal’s corpuscles in the thymus. At the DP (double-positive) stage, they are
selected by their interaction with the cells within the thymus, begin the transcription of Foxp3, and
become Treg cells.
Tregs have a larger TCR diversity than effector T cells, biased towards self-peptides.
The process of Treg selection is determined by the affinity of interaction with the self-peptide MHC
complex. Selection to become a Treg is a “Goldilocks” process - i.e. not too high, not too low, but just
right; a T cell that receives very strong signals will undergo apoptotic death; a cell that receives a weak
signal will survive and be selected to become an effector cell, If a T cell receives an intermediate signal,
then it will become a regulatory cell.
Due to the stochastic nature of the process of T cell activation, all T cell populations with a given TCR
will end up with a mixture of Teff and Treg – the relative proportions determined by the affinities of the
T cell for the self-peptide-MHC.
Foxp3+ Treg generation in the thymus is delayed by several days compared to Teff cells and does not
reach adult levels in either the thymus or periphery until around three weeks post-partum. Treg cells
require CD28 co-stimulation and B7.2 expression is largely restricted to the medulla, the development of
which seems to parallel the development of Foxp3+ cells.
Molecular characterization
Expression of Foxp3 is required for regulatory T cell development and appears to control a genetic
program specifying this cell's fate.
The large majority of Foxp3-expressing regulatory T cells are found within the major histocompatibility
complex (MHC) class II restricted CD4-expressing (CD4+) population and express high levels of the
interleukin-2 receptor alpha chain (CD25). In addition to the Foxp3-expressing CD4+ CD25+, there also
appears to be a minor population of MHC class I restricted CD8+ Foxp3-expressing regulatory T cells
but not in case of a healthy person but in case of autoimmune disorder.
Function
the following represent some of the proposed mechanisms of immune suppression:
 Regulatory T cells produce a number of inhibitory cytokines. These include Transforming
growth factor beta,[14] Interleukin 35,[15] and Interleukin 10.[16] It also appears that regulatory T cells can
induce other cell types to express interleukin-10.[17]
 Regulatory T cells can produce Granzyme B, which in turn can induce apoptosis of effector
cells. Regulatory T cells from Granzyme B deficient mice are reported to be less effective suppressors of
the activation of effector T cells.
 Reverse signalling through direct interaction with dendritic cells and the induction of
immunsuppressive indoleamine 2,3-dioxygenase.
 Signalling through the ectoenzymes CD39 and CD73 with the production of
immunosuppressive adenosine.
 Through direct interactions with dendritic cells by LAG3 and by TIGIT.
 Another control mechanism is through the IL-2 feedback loop. Antigen-activated T cells produce
IL-2 which then acts on IL-2 receptors on regulatory T cells alerting them to the fact that high T cell
activity is occurring in the region, and they mount a suppressory response against them. This is a
negative feedback loop to ensure that overreaction is not occurring. If an actual infection is present other
inflammatory factors downregulate the suppression. Disruption of the loop leads to hyperreactivity,
regulation can modify the strength of the immune response. [24] A related suggestion with regard
to interleukin 2 is that activated regulatory T cells take up interleukin 2 so avidly that they deprive
effector T cells of sufficient to avoid apoptosis.
 A major mechanism of suppression by regulatory T cells is through the prevention of co-
stimulation through CD28 on effector T cells by the action of the molecule CTLA-4.
INDUCED REGULATED T CELL
Induced regulatory T (iTreg) cells (CD4+ CD25+ Foxp3+) are suppressive cells involved in tolerance.
iTreg cells have been shown to suppress T cell proliferation and experimental autoimmune diseases.
These cells include Treg17 cells. iTreg cells develop from mature CD4+ conventional T cells outside of
the thymus: a defining distinction between natural regulatory T (nTreg) cells and iTreg cells.
Acute depletion of the iTreg cell pool in mouse models has resulted in inflammation and weight loss.
Epigenetic differences have been observed between nTreg and iTreg cells, with the former having more
stable Foxp3 expression and wider demethylation.
The small intestinal environment is high in vitamin A and is a location where retinoic acid is produced.
The retinoic acid and TGF-beta produced by dendritic cells within this area signal for production of
regulatory T cells.
Vitamin A and TGF-beta promote T cell differentiation into regulatory T cells opposed to Th17 cells,
even in the presence of IL-6.
Some of the itregs express the lectin-like receptor CD161 and are specialized to maintain barrier
integrity by accelerating wound healing.
The Tregs within the gut are differentiated from naïve T cells after antigen is introduced.
DISEASES
While the immunosuppressive function of regulatory T cells prevents the development of autoimmune
disease, it is not desirable during immune responses to infectious microorganisms.
Experimental evidence from mouse models suggests that some pathogens may have evolved to
manipulate regulatory T cells to immunosuppress the host and so potentiate their own survival. For
example, regulatory T cell activity has been reported to increase in several infectious contexts, such as
retroviral infections (the most well-known of which is HIV), mycobacterial infections (like
tuberculosis), and various parasitic infections including Leishmania and malaria.
Tregs play major roles during HIV infection. They suppress the immune system, thus limiting target
cells and reducing inflammation, but this simultaneously disrupts the clearance of virus by the cell-
mediated immune response and enhances the reservoir by pushing CD4+ T cells to a resting state,
including infected cells. Additionally, Tregs can be infected by HIV, increasing the size of the HIV
reservoir directly.
Regulatory T cells have a large role in the pathology of visceral leishmaniasis and in preventing excess
inflammation in patients cured of visceral leishmaniasis.
CD4+ regulatory T cells are often associated with solid tumours in both humans and murine models.
Increased numbers of regulatory T cells in breast, colorectal and ovarian cancers is associated with a
poorer prognosis.
CD70+ non-Hodgkin lymphoma B cells induce Foxp3 expression and regulatory function in intra
tumoral CD4+CD25− T cells.
A recent study shows that cerebral ischemia can increase bone marrow CD4(+)CD25(+)Foxp3(+)
regulatory T cells via signals from the sympathetic nervous system.
There is some evidence that Tregs may be dysfunctional and driving neuroinflammation in amyotrophic
lateral sclerosis due to lower expression of Foxp3.
CANCER
Most tumors elicit an immune response in the host that is mediated by tumor antigens, thus
distinguishing the tumor from other non-cancerous cells. This causes large numbers of tumor-infiltrating
lymphocytes (TILs) to be found in the tumor micro environment.
While Tregs normally make up only about 4% of CD4+ T cells, they can make up as much as 20–30%
of the total CD4+ population around the tumor microenvironment.
The ratio of Tregs to T effectors in the tumor microenvironment is a determining factor in the success of
the immune response against the cancer. High levels of Tregs in the tumor microenvironment are
associated with poor prognosis in many cancers, such as ovarian, breast, renal, and pancreatic cancer.
This indicates that Tregs suppress Teffector cells and hinder the body's immune response against the
cancer.
However, in some types of cancer the opposite is true, and high levels of Tregs are associated with a
positive prognosis. This trend is seen in cancers such as colorectal carcinoma and follicular lymphoma.
This could be due to Treg's ability to suppress general inflammation which is known to trigger cell
proliferation and metastasis.
The chemotaxis is probably driven by the production of chemokines by the tumor. Treg infiltration into
the tumor microenvironment is facilitated by the binding of the chemokine receptor CCR4, which is
expressed on Tregs, to its ligand CCL22, which is secreted by many types of tumor cells.
The cytokine, TGF-β, which is commonly produced by tumor cells, is known to induce the
differentiation and expansion of Tregs.
Depletion of Tregs in animal models has shown an increased efficacy of immunotherapy treatments, and
therefore, many immunotherapy treatments are now incorporating Treg depletion.
FUNCTIONS OF OTHER T CELL SUBSETS
γδ T cells and NKT cells recognize a wide variety of antigens, many of which are not peptides, and these
are not displayed by class I and class II MHC molecules on APCs.
The antigen receptors of many γδ T cells and NKT cells have limited diversity, suggesting that both cell
types may have evolved to recognize a small group of microbes. Because of this feature, these T cells
are often said to be at the crossroads of innate and adaptive immunity.
Both cell types are abundant in epithelial tissues, such as the gastrointestinal tract.
γδ T Cells
The antigen receptor of MHC-restricted CD4+ and CD8+ T lymphocytes is a heterodimer composed of
α and β chain. There is a second type of clonally distributed receptor composed of heterodimers of γ and
δ chains, called γδ T cell.
The percentages of γδ T cells is less than 5% of all T cells express this form of TCR.
The γδ heterodimer associates with the CD3 and ζ proteins in the same way as TCR αβ heterodimers do.
The limited diversity of the γδ TCRs in many tissues suggests that the ligands for these receptors may be
invariant an conserved.
One intriguing feature of γδ T cells is their abundance in epithelial tissues of certain species. For
example, more than 50% of lymphocytes in the small bowel mucosa of mice and chickens, called
intraepithelial lymphocytes, are γδ T cells.
In mouse skin, most of the intraepidermal T cells expres the γδ receptor, only about 10% of human
intestinal intraepithelial T cells express the γδ TCR.
Other γδ T cells recognize protein or nonprotein antigens that do not require processing or any particular
type of APCs for their presentation.
Many γδ T cells are triggered by microbial heat shock proteins.
It has been postulated that this subset of T cells may initiate immune responses to microbes at epithelia,
before the recruitment and activation of antigen-specific αβ T cells. However, mice lacking γδ T cells,
created by targeted disruption of the γ or δ TCR gene, have little or no immunodeficiency.
NKT Cells
A small population of T cells also expresses markers that are found on NK cells, such as CD56; these
are called NKT cells.
The TCR α chains expressed by a subset of NKT cells have limited diversity, and in humans, these cells
are characterized by a V region encoded by a rearranged Vα24-Jα18 gene segment, with little or no
junctional diversity, associated with one of three β chains. Because of this limited diversity, these cells
are also called invariant NKT (iNKT) cells.
All NKT cell TCRs recognize lipids that are bound to class I MHC–like molecules called CD1
molecules.
NKT cells and other lipid antigen– specific T cells are capable of rapidly producing cytokines such as
IL-4 and IFN-γ after activation, and they may help marginal zone B cells to produce antibodies against
lipid antigens.
NKT cells may mediate protective innate immune responses against some pathogens, such as
mycobacteria.