T CELLS AND T CELL-MEDIATED IMMUNITY

DEFINITION

T cells are lymphocytes that express TCRs and develop in the thymus. They have different subsets
and are involved in cell-mediated adaptive immunity.

They develop from lymphocyte precursors, which go into the thymus and differentiate into two
major subsets of naïve T cells which may possess CD3/CD4 or CD3/CD8 markers. Naïve T cells then are
activated by antigen presentation and divide into effectors and memory T cells. Naïve T cells can be found in
secondary lymphatic organs, as they enter the lymphocyte recirculation after differentiation. 2/3 of the T
cells will be Th cells (CD3/CD4, they activate B cells) and 1/3 Tc cells (CD3/CD8, they kill target cells).

T cells have TCR complexes on their surfaces which are heterodimers and recognise antigens bound
to MHCs. They rely on CD3 complexes (heterodimer too) to transduce the signals. 95% of TCR complexes are
made of alpha and beta subunits, while the remaining 5% are gamma and delta.

DEVELOPMENT

The role of the thymus is to ensure that only self-MHC restricted T cells that are unable to react with
self-antigens can get to the periphery. The aim of T cell differentiation is to:

 Ensure self-MHC recognition by TCR

 Remove unnecessary clones
 Remove self-endangering clones
 Ensure CENTRAL TOLERANCE - Central tolerance is the mechanism by which newly developing T cells and B cells are rendered non-
reactive to self, it occurs while developing immune cells are still present in the primary lymphoid organs (the thymus and bone-marrow), prior to export into
the periphery
 Develop TCR repertoire that can recognise foreign antigens
 Ensure individualised optimal T cell repertoire in the periphery

The stroma of the thymus provides a microenvironment that ensures T cell development and
selection. Pro-T cells reach the thymus from the bone marrow. Inside the thymus they first undergo IL-7-
dependent proliferation, together with beta-chain rearrangement; this happens in two steps:
1) D-J rearrangement, resulting in a CD25+CD44 T cell;
2) V-DJ rearrangement, resulting in a CD25+CD44 T cell with intracellular beta-chains. At the end of this step
the cell can:

 Be destroyed because of an unsuccessful beta-chain;

 Become a delta/gamma T cell (they undergo no selection);
 Become an early pre-T cell;

Early pre-T cells will express TCRs composed of beta-chains with surrogate alpha-chains and very low CD3;
these pre-alphaTCR complexes will inhibit further beta-chain rearrangement (guaranteeing antigen-
specificity) and will induce them to undergo further development by proliferation, expression of CD4/CD8,
and transcription and V-J rearrangement of alpha-chains, which will give CD4 and CD8 positive (DOUBLE
POSITIVE) cells called late pre-T cells.

DOUBLE POSITIVE T cells will be selected by positive selection in the cortex (using epithelial cells)
and negative selection in the corticomedullary region (using DCs and macrophages):

 During positive selection, the cells will become single positive, depending on whether they
recognise MHCI or MHCII molecules; those that will not recognise MHC will undergo apoptosis
(SELECTION).
 After that, the cells will be negatively selected: if they interact tightly with self-antigens, they will
undergo apoptosis (SELECTION); if NOT (=NEGATIVE) they will be allowed to live.The negative
selection CANNOT eliminate T cells whose receptors are specific for self-antigens that are expressed
outside of the thymus (these t cells do not yet have peripheral tolerance as compared to central
tolerance acquired inside the thymus); they will enter circulation, but will be soon rendered anergic
or unresponsive by other mechanisms.

The thymus will express ubiquitous cell-associated (COMMON TO EVERY CELL), circulating proteins, and
tissue-specific antigens (TSAs); their expression in medullary thymic epithelial cells (mTECs) is controlled by
AIRE. AIRE gene is expressed not only in the thymus, but also in many other tissues as well. CENTRAL
TOLERANCE is achieved in this way, as the T cells are made non-reactive to self in the thymus, a primary
(central) lymphoid organ; it is opposed to peripheral tolerance, where the T cells that are reactive to
TSAs(Tissue Specific Antigens) which may have escaped negative selection, are stopped in the peripheral
tissues by other mechanisms:

 SUPPRESSION by regulator T cells;

 ANERGIC response to APCs;
 DELETION responses to APCs – T cells are induced to apoptosis by APCs.
 FUNCTION

There are several types of T cells with several functions:

 T helper cells
 T cytotoxic cells
 Gamma/delta T cells
 Regulatory T cells
 NKT cells

T helper cells: there are mainly five types:

 Induced T reg: TGF-beta in regulation/suppression of immune/inflammatory responses

 TH17: IL1, IL6, IL23, TGF-beta; IL17, IL17F, IL22 in inflammation
 TH2: IL4 in allergic and anti-helminth response
 TH1: INF-gamma in cell-mediated immunity
 TFH: IL21 in B cell help in germinal centres

Cytotoxic T cells are used in cell-mediated immunity. Once activated, they can kill the infected cells by
inducing apoptosis (Fas-FasL interaction; TNF-alpha) or by inducing cell lysis (perforins; granzymes).

Activation of naïve Th and Tc cells is analogous; it occurs when the TCR-CD3 complex binds to the
MHC-antigen complex. Their binding is weak, therefore accessory molecules are required (CD4 and CD8) to
stabilise the bond. To initiate the activation however, this interaction itself is not enough; costimulatory
signals are needed by the naïve T cells. This is mediated by CD80/86 or CD40 molecules on the APC that
bind to CD28 or CD40L molecules on the naive T cell.

The TCR complex is composed of the alpha/beta dimer receptor and a co-receptor. The T-cell co-
receptor is composed of four distinct chains:1 CD3γ chain +1 CD3δ chain +2 CD3ε chains; ζ-chains generate
the activation signal in T lymphocytes.

When an APC forms a synapse with a T cell, the latter will be activated: the TCR complex and the
accessory molecules will interact with the MHC-antigen complex; the CD28 (cytotoxic) or CD40L (helper) co-
receptors will interact with B7-1 (=CD80) or B7-2 (=CD86) and CD40, respectively. The interaction between
the MHC and the TCR complexes will result in a series of reaction: on the TCR complex the fynprotein kinase
will be activated, together with the Lck protein kinase which is associated to the accessory molecules; upon
their activation, fyn and Lck will phosphorylate tyrosine residues on theζ-chains. This will lead to binding of
ZAP-70, which will phosphorylate other molecules, inducing the phosphatidylinositol pathway activation,
leading to intracellular calcium concentration increase, which will cause translocation of NF-AT into the
nucleus; this molecule will upregulate IL2 alpha-chain gene expression. This will cause the production of the
alpha-chain of the IL2 receptor, which until then was composed of only gamma and beta-chains, rendering
it poorly affine to IL2; this is also called the SIGNAL 1 of T cell activation.

However, the binding of MHC itself is not sufficient for T cell activation; in fact, lack of further stimulation
leads the T cell into ANERGY: cells are unable to proliferate or be clonally selected. The costimulatory signal
necessary to continue the immune response can come from B7-CD28 and CD40-CD40L interaction. These
are the sources of SIGNAL 2. This signal with activate AP-1 and NFkappa-B to increase IL2 production by
100x overall. Cytokines could ulteriorly contribute to the activation: SIGNAL 3. Immunosuppressive drugs
illustrate the importance of IL-2 in immune responses: e.g.Cyclosporin inhibits IL-2 by disrupting TCR
signalling;Rapamycin inhibits IL-2R signalling. Costimulation however, does not only enhance IL-2
production but also induces expression of CTLA-4 receptors; these will bind B7 molecules with a higher
affinity then CD28 and will induce a negative feedback inhibition of IL-2 production, it acts as a brake (in
antitumor therapy, it will be inhibited by IPILIMUMAB, to keep the T cells activated, and prevent their
inhibition; in case of Hodgkin’s or non-Hodgkin’s lymphomas, related to B cells over activation,
LUCATUMUMAB is used, to inhibit CD40 interaction with malignant B cells and mediate ADCC).

Gamma/delta T cells: they have multiple ligands, which allow them to interact almost universally with the
components of the immune system and against the pathogenic microbes or molecules. Mostly, we
distinguish three types:

1. MHC-related and -unrelated T cell receptor (TCR) ligands ;

2. various cell surface molecules that engage the activating natural killer receptors (NKRs);

3. DAMPs and PAMPs receptors (=PRRs).

These cells are generally used to suppress the immune responses, by local immunoregulation: they block
neutrophils, induce cytolysis and general suppression of effector T cells, local APCs and epithelial cells,
antagonise TH-1 activity, and induces reinforcement of the epithelial cell barrier.

DETECTION

You can detect them by morphology (flow cytometry) or function (response to antigen by
proliferation assays or cytokine production detection); cytotoxicity assays.

DISEASES

Remember three of them:

 THYMIC APLASIA/HYPOPLASIA or DIGEORGE SYNDROME– missing thymus

 HYPER IGM SYNDROME– missing CD40L leads to inability to produce antibodies other than IgMs
 AIDS (=HIV INFECTION)– virus.