Introduction

Where do the T cells develop? Predominantly in thymus

Where is thymus Located? The ventral part of third pharyngeal pouch. Endoderm is the only site from which thymus primordium emerges

Note : the dorsal part gives rise to the parathyroid gland

The Thymus
Lobulated structure with a STROMA of epithelial cells & connective tissue. Stroma provides a microenvironment for T cell development & selection. Lobules differentiated into an outer CORTEX and Inner MEDULLA, both filled with bone marrow derived THYMOCYTES

What is the evidence that the thymus is involved in the generation of T cells?
Thymectomy- adult versus newborn Athymic (nu/nu)- no hair cum thymic deficiency Mutation in whn (winged helix nude). A Forkhead transcription factor known as FOXN 1 which is expressed in thymic epithelial cells, hair follicles and skin epithelial cells, causes the nude phenotype. It is required for the normal differentiation of hair follicles and thymus epithelial cells

Digeorge syndrome - developmental defect ( deletions in chromosomes 22q11) causes immunodeficiency along with hypoparathyoidism and congenital heart disease due to mutations in the T-box transciption factor known as TBx1 Note: vitamin A deficiency and excess retinoic acids reduces TBx1. Thus it is important to note that proper amount of Vitamin A and retinoic acids during development are important for expression of TBx1 and formation of t cell in thymus

T cells T cells
Some contain invariant TCR whereas other contain variable TCRs and innate role Immunomodulation = reducing inflammation/ Injury of epithelial cells ( as a result of an Immune response, eg after infection or allery). By secreting epithelial cell growth factors, T cells enhance the recovery of epithelial cells after an immune response. No need for MHC or MHC processing pathways. Some evidence that MHC class b molecules may be up-regulated after activation, stress, transformation etc leading to recognition by T cells. T cells kills intestinal cells expressing MICA/MICB.

gd T cells are favored during early fetal development

Antigen recognition by gd T cells is different than T cells

gd T cells are not MHC restricted!

Antigen is recognized directly, more like an antibody

In some cases ligands for the gd TCR are self proteins upregulated under stress conditions In humans, circulating gd cells recognize a phospholipid antigen from Mycobacterium tuberculosis

T cells T cells
CD4 or CD8 TCR recognises self MHC+ peptide, therefore, TCR should recognise self MHC but should NOT recognise self antigens

Methods of study
Use antibodies to cell surface molecules & FACS Use bone marrow chimeras or fetal thymic organ cultures (FTOC) or transgenic TCR mice or mice without key immune molecules, athymic mice, Rag 1 & 2 deficient or SCID mice ( lack DNA dependent protein kinase involved in repair of ds DNA breaks)

T Cell education
Bone marrow

Primitive T cell
Positive selection Selects T cells that can bind to MHC. Removes cells that cannot be recognised Negative selection Removes strongly self Reactive T cells Mature T cells

Apoptosis

Early Development
What is required for maturation in the thymus? Experiment by J.C. Zuniga Pfluker: Gene knockout on mice T Cells can develop in the absence of thymic fragments Notch1 ligand required for T-cell precursor to form

Developmental Steps in the Thymus

Cell-Surface Molecules

c-Kit: Receptor for stem cell growith factor CD44: Adhesion Molecule CD25: Alpha-Chain of the IL-2 receptor

T-Cell Development

T-Cell precursor in Double Negative State. DN1 DN4 changes in cell-surface protein expression. DN1 is a Thymic progenitors. DN2 is a crucial state RAG expression gene rearrangements Determination of beta, gamma and delta Chain of TCR CD3 gene is expressed

In DN4 stage, TCR-beta is expressed together with pT-, signaling by this TCR results in increased survival, proliferation and differentiation. Resulting in expression of CD* followed by CD4, i.e. cells become DP

TCR is now in the Double Positive (DP) State

Thymic Selection Processes

Thymocytes are in the Double Positive State (i.e. express both coreceptors. Positive and Negative Selection necessary to produce an effective T-Cell repertoire.

Positive Selection
Results in MHC restriction Mechanism:
Immature thymocytes cluster with MHC molecules on the cortical cells of the thymus
If TCR interacts with MHC protective signal results that prevents apoptosis. If TCR does not interact with MHC no protective signal and apoptosis occurs.

Result? Only reactive thymocytes survive.

Negative Selection
Ensures self-tolerance Weeds out High affinity thymocytes Mechanism:
APCs bearing MHCs interact with thymocytes
If avidity is too strong thymocyte undergoes apoptosis. Details unknown

Result? Only self-tolerant thymocytes survive.

Positive Selection Experiment

Thymocytes are dependent on the presence of MHCs in the thymus for survival. Class I MHC knockout CD4 is selected for, but CD8 does not survive. Class II MHC knockout CD8 is selected for, but CD4 does not survive.