Unit 8

T-Cell Maturation, Activation,

and Differentiation
T-Cell Maturation, Activation, and Differation

 Tcell maturation requires rearrangements of the germ-line, TCR genes,

and the expression of various membrane markers.
 Developing T cells, known as thymocytes, proliferate and differentiate along
developmental pathways
 Functionally distinct subpopulations of mature T cells develop...
 Final maturation of most T cells proceed along two different
developmental pathways, which generate functionally distinct
subpopulations
 CD4 exhibiting class II MHC restriction, and
 CD8 exhibiting class I MHC restriction.
Germline rearrangement of alpha and beta
chain gene and expression
T-Cell Maturation and the Thymus

 Thymus main source of all T cells, it is where T cells diversify and are
shaped into an effective primary T-cell range by extraordinary selection
processes.
 positive selection permits survival of only T cells whose TCRs are capable of
recognizing self-MHC molecules. It is responsible for creation of a self-MHC-
restricted repertoire of T cells.
 negative selection, eliminates T cells that react too strongly with self-MHC or with
self-MHC plus self peptides. It is an extremely important factor in generating T-cells
that are self-tolerant.
 With this maturation the lymphocytes are CD3+
T cell maturation

Migration of Stem cells to the Migration of Thymocytes to

Thymus early in development secondary lymphoid organs
and maturation in to after maturation for interaction
thymocytes (T cells) with potential antigens
Thymic Selection of the T-Cell (Range) Repertoire

 Random gene rearrangement within TCR germ-line DNA combined with

junctional diversity can generate an enormous TCR repertoire,
 with an estimated potential diversity exceeding
 1015 for the TCR receptor and
 1018 for the  TCR receptor.
 they should be capable of recognizing
 Soluble antigen (either foreign or self), self-MHC molecules,or antigen plus a nonself-MHC
molecule.
 the most distinctive property of mature T cells is that they recognize only
foreign antigen combined with self-MHC molecules.
Conti….

 Thymocytes undergo two selection processes in the thymus:

 Positive selection for thymocytes bearing receptors capable of binding
self-MHC molecules, which results in MHC restriction.
 Cells that fail positive selection are eliminated within the thymus by apoptosis (≥98%
before leave thymus).
 Negative selection that eliminates thymocytes bearing high-affinity
receptors for self-MHC molecules alone or self-antigen presented by self-
MHC, which results in self-tolerance.
 Steps in T cell development

Step 1. Positive selection

occurs in the thymic cortex

MHC self-
recognition
molecules
Steps in T cell development (cont’d)

Step 2. Negative selection

occurs in the thymic
medulla.
Structure of the T cell Receptor
 Heterodimer with one
α and one β chain of
roughly equal length
 A short cytoplasmic
tail
 A transmembrane
region with
hydrophobic amino
acids
Structure of the T cell Receptor
 Both α and β chains have
a variable (V) and
constant (C) region
 V regions of the α and β
chains contain
hypervariable regions that
determine the specificity
for antigen
γδ TCR
 Small population of T cells express a TCR that
contain γ and δ chains instead of α and β chains
 The Gamma/Delta T cells predominate in the
mucosal epithelia
 Gamma/Delta T cells play a role in responses to
certain viral and bacterial pathogens
 Genes for the δ chains have V, D and J gene
segments; γ chains have V and J gene
segments
 Repertoire is limited
T-Cell Receptors
 The T-cell receptor (TCR) associates with
CD3, forming the TCR-CD3 membrane
complex.
 The accessory molecule participates in
signal transduction after interaction of T cell
with antigen;
 it does not influence interaction with antigen.
 CD3 is closely to the  heterodimer and is
required for membrane expression of  and 
T-cell receptors
T-Cell Receptors
 Each heterodimer forms a complex with CD3 on
the T-cell membrane. The T-cell receptor
complex can be envisioned as four dimers:
 the  or  TCR heterodimer determines the ligand-
binding specificity,
 the CD3 dimers: gamma-epsilon () , delta-epsilon
(), and 2 zeta, are required for membrane
expression of T-cell receptor and for signal
transduction

 Class I MHC–specific TCR together with the

CD8 co-receptor generates a different signal
 Class II MHC–specific TCR together with the
CD4 co-receptor also produces a different
signal.
T-Cell Accessory Membrane Molecules

 CD4 and CD8 are classified as coreceptors based on their

abilities to recognize the peptide-MHC complex and their roles in
signal transduction.
 The extracellular domains of CD4 and CD8 bind to the conserved
regions of MHC molecules on antigen-presenting cells (APCs) or
target cells.
TCR and CD3 Complex

 TCR is closely
associated with a
group of 5 proteins
collectively called the
CD3 complex
 γ chain
 δ chain
 2 ε chains
 2 ξ chains
 CD3 proteins are
invariant
Role of CD3 Complex

 CD3 complex
necessary for cell
surface expression of
TCR during T cell
development

 CD3 complex
transduces signals to
the interior of the cells
following interaction of
Ag with the TCR
The “Immunological Synapse”
 The interaction between
the TCR and MHC
molecules are not strong
 Accessory molecules
stabilize the interaction
 CD4/Class II MHC or
CD8/Class I MHC
 CD2/LFA-3
 LFA-1/ICAM-1
The “Immunological Synapse”
 Specificity for antigen
resides solely in the TCR
 The accessory molecules
are invariant
 Expression is increased in
response to cytokines
The “Immunological Synapse”
 Engagement of TCR and Ag/MHC is
one signal needed for activation of T
cells
 Second signal comes from
costimulatory molecules
 CD28 on T cells interacting
with B7-1 (CD80) or B7-2
(CD86)
 Others
 Costimulatory molecules are invariant
 “Immunological synapse”
Costimulation is Necessary for T Cell Activation

 Engagement of TCR and Ag/MHC

in the absence of costimulation can
lead to anergy
 Engagement of costimulatory
molecules in the absenece of TCR
engagement results in no response
 Activation only occurs when both
TCR and costimulatory molecules
are engaged with their respective
ligands
 Downregulation occurs if CTLA-4
interacts with B7
 CTLA-4 send inhibitory signal
TH-Cell Activation
 TH cell activation is initiated by interaction of the TCR-
CD3 complex with a processed antigenic peptide bound
to a class II MHC molecule on the surface of an antigen-
presenting cell - APC.
 Interaction of a TH cell with antigen initiates a cascade of
biochemical events that induces the resting TH cell to
enter the cell cycle, proliferating and differentiating into
memory cells or effector cells.
 Resulting in release of specific cytokines and forming
adhesion molecules.
Key Steps in T cell Activation

 APC must process and present peptides to T cells

 T cells must receive a costimulatory signal
 Usually from CD28/B7
 Accessory adhesion molecules help to stabilize binding of T cell and APC
 CD4/MHC-class II or CD8/MHC class I
 LFA-1/ICAM-1
 CD2/LFA-3
 Signal from cell surface is transmitted to nucleus
 Second messengers
 Cytokines produced to help drive cell division
 IL-2 and others
T-Cell Differentiation

 Niave T cells that have not encounted an antigen-MHC complex

repeatedly migrate through the circulation and lymphatics.
 When these cells encounter an antigen-MHC complex they
undergo repeated cell division producing effector and memory
cells.
Cell Death and T-Cell Populations

 is an important feature of lymphocyte

homeostasis, returning T- and B-cell populations
to their appropriate levels after bursts of antigen-
induced proliferation.
 Apoptosis also plays a crucial role in the deletion
of potentially autoreactive thymocytes during
negative selection and in the removal of
developing T cells unable to recognize self
(failure to undergo positive selection).
Cell Death and T-Cell Populations

 Outside of the thymus, most of the TCR-mediated

apoptosis of mature T cells is mediated by the Fas
pathway.
 Repeated or persistent stimulation of peripheral T
cells results in the coexpression of both Fas and Fas
ligand, followed by the apoptotic death of the cell.
 The Fas/FasL mediated death of T cells as a
consequence of activation is called activation-induced
cell death (AICD) .
 The T-cell repertoire is shaped by apoptosis in the
thymus and periphery.
Cont…
Signal Transduction by the TCR Complex

 Recognition of Ag by TCR initiates a sequence of biochemical signals in T

cells
 Goal is to coordinately activate the transcription of genes that are silent in
naive cells and whose products mediate the responses and functions of
activated T cells.
 Main biochemical pathways activated includes
 Ras-MAP kinase pathway,
 the protein kinase C pathway,
 calcium-calcineurin pathway
 Each of these pathways contributes to the expression of genes encoding
proteins needed for T cell clonal expansion, differentiation, and effector
functions.
Intracellular signaling events during T cell activation
Superantigen induced T-cell activation
 Superantigens:
 viral or bacterial peptides that
can simultaneously bind to the
V domain of TCR and -chain
of a class II MHC molecule,
outside the peptide-binding site
 a whole 'family' of T cells
respond, rather than a single
clone, with excessive and
potentially damaging over-
production of cytokines.
Conti…..

 Both exogenous and endogenous superantigens have been

identified.
 Crosslinkage of a TCR receptor and class II MHC molecule by
either type of superantigen produces an activating signal that
induces T-cell activation and proliferation
 Exogenous superantigens are soluble proteins secreted by
bacteria
 among them are a variety of exotoxins secreted by gram-
positive bacteria, such as staphylococcal enterotoxins, toxic-
shock-syndrome toxin, and exfoliative-dermatitis toxin.
Conti…
 Each of these exogenous superantigens binds particular V
sequences in T-cell receptors and crosslinks the TCR to a class
II MHC molecule.
 The massive activations that follow crosslinkage by a
superantigen results in overproduction of TH-cell cytokines,
leading to systemic toxicity.
 Example:
 food poisoning: induced by staphylococcal enterotoxins
 toxic shock: induced by toxic shock-syndrome toxin-1 (TSST-
1).
Conti…

 Endogenous superantigens
 are membrane-embedded proteins produced by certain viruses
that infect mammalian cells;
 they include Mls antigens (minor lymphocyte stimulating)
encoded by mouse mammary tumor virus (MTV).