Th cell Activation and differentiation

• 1. Activation
– TCR coupled signaling pathways
– Co-stimulatory signal-Differences amongst APC’s
– Super antigen induced Tcell activation

• 2. Differentiation
– Generation of Effector and Memory cells
– Cell death of Tcell population
– Function of peripheral gamma, delta T cells
 Concept
• Interaction of TCR CD3 complex
+
• Processed Antigenic peptide bound MHC I or II
• Signals
• Cascade of Events
• Proliferation
• Differentiation: Memory cell
Effector cells
 Gene Products
• Immediate genes:30 min
Transcription factors: cFos, cMyc,NF-AT

• Early Genes:1-2hrs
Various Cytokines: Il-2,IL-2R,IL-3 etc

• Late Genes-2 days

Adhesion molecules VLA-4,HLA-DM
 Common themes in signal Transduction: Environmental Stimuli

• Receptor: On cell surface

• Intracellular- hydrophobic ex: steroids
• Generation of second messenger:diffuse into other sites of cell
eg:cAMP,cGMP,Ca,DAG,IP3
• Protein Kinases :add Phosphate
• Protein Phosphatases Remove Phosphate tyrosine, serine or
threonine residues
• Induced Assembly: Adaptor proteins sites for receptor and
cytoplasmic molecule
• Large and small G proteins :GTP activates GDP inactivates
• Cascade of Events
• Default setting is Off
 TCR coupled signaling pathways
• Fyn protein tyrosine kinase: near tail of TCR CD3 complex
• Translocation of Co receptors near TCR
• p56Lck protein tyrosine kinase (bought near TCR CD3 tail)
• Phosphorylate Immunoreceptor Tyrosine Based Activation Motif ITAM
(CD3)
• Creates Docking site for SH2 site of ZAP-70
• This activates Many pathways
• Phospholipase C gamma splits phosphatidylinositol 4,5-biphosphate
into Inositol 1,4,5-Triphosphate and DiacylGlycerol
• IP3- increases Ca+2 --- activates Calmodulin dependent Phosphatase
called Calcineurin—dephosphorylates inactive form NF-AT
• DAG activates Protein Kinase PKC which phosphorylates and releases
Nuclear factor Kappa B
 Co-stimulatory signal-Differences amongst
APC’s
• Signal 1: TCR+ Ag peptide signal cascade
• Signal 2:Ag non specific
– Interaction b/w CD28(t cell) & B7 fly(APC)
– B7-1,B7-2 Ig superfly
– V & c like domain ,cytosolic domains diff

– Constitutively expressed : Dc
– Induced on activated macrophages,B cells

– Ligand for B7:CD28

» CTLA-4 Cytotoxic T Lymphocyte AssociatedProtein
4
 Lymphocyte Homeostasis
• CD28: + • CTLA-4: negative
• T cell proliferation • Downregulatory
• P on resting • P on activated cells
&activated cells . • Detected 24hrs
• Peaks high • Peak 2-3 days
• Less affinity a • Higher affinity
• As its level increases • In Knockout mice:
CTLA level go up
• Lymphadenopathy
• Regulatory feedback
• splenomegaly
 Super antigen induced Tactivation
• Viral/bacterial Ag
• Vbeta domain of TCR & alpha chain of MHC II
• Exogenous: Staphylococcal enterotoxin
Toxic shock syndrome
• Endogenous: cm ptns. Retroviral ptns called Minor
lymphocyte stimulating determinants produced my
Mouse Mammary Tumor virus.
• Any T cell with particular Vbeta domain: polyclonal
activation: over production of Cytokines
• Influence T cell maturation in Thymus : T cells lost by
negative selection
 Generation of Effectors and Memory cells
• CD4/8 (naïve cells) leave thymus: enter
circulation(G0):recirculate blood lymph node till it
encounters Ag:12-24hrs
• After encounters Ag
• TCR CD3 coupled signal
• Co stimulatory signal
• Blast cell
• G1 phase
• IL-2 production 100 fold
• Activated cells divide 2-3 for 4-5 days
• Differentiate into memory effectors cells
 Death of Tcell : APOPTOSIS
• Programmed Cell death
• Lymphocyte Homeostasis
• Deletion of autoreactive thymocytes in Negative
selection
• Proteases called “CASPASES”
• 14 mammals ,highly conserved
• Every cell posses it
• Caenorhabditis elegans
• Cysteine residues specificity for asparatic acid
residues
 Two Pathways
• Ag stimulation increases • Negative Selection
• Fas • Cytochrome c leaks from
• FasL mitochondria
• FADD • Binds Apaf-1 Apoptopic
• Procaspase 8 Protease Activating factor
• Caspase 8 • Procaspase 9
• Procaspase 3 • Cyt3+Apaf-
• Caspase 3 1+Procaspase1:
APOPTOSOME
• Proteolysis essential
• Caspase 9
molecules
• Caspase 3
• AICD Activation Induced
Cell Death • Bid 3: Activates
Mitochondrial pathway
 Function of peripheral gamma, delta T cells

• 5% Skin, intestinal • Imp role in immune

• pulmonary epithelium response: location
• Some express only CD3 commonly encounters Ag
• Primary line of defense
• Some CD3+CD8
• Non peptide Ag
• No recirculation
• Conserved epitopes on
• React with Ag found at
Ag
these sites.
• Binds : ptn lik Abs • Clearance damaged cells
• No clear cut
Not commonly bound to
APCs • Tumor cell lysis