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Cytotoxic T cell. Helper T cell. T H. Tc. T cell receptor. Fragments of foreign protein. Class II MHC protein. Class I MHC protein. Fragments of foreign protein. Antigen presenting cell. infected target cell. Helper T cells: • Express CD4 molecule on the cell surface.

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Presentation Transcript
slide1

Cytotoxic T cell

Helper T cell

TH

Tc

T cell receptor

Fragments

of foreign

protein

Class II

MHC

protein

Class I

MHC

protein

Fragments

of foreign

protein

Antigen

presenting cell

infected

target cell

slide2

Helper T cells:

• Express CD4 molecule on the cell surface.

• Recognize antigen peptides bound to class II MHC molecules (exogenous or ingested antigens).

• Provide help to B cells, macrophages and other T cell effector responses though the production of cytokines

Cytotoxic T cells:

• Express CD8 molecule on the cell surface.

• Recognize antigen peptides bound to class I MHC molecules (endogenously produced antigen such as viral antigen).

• Kill infected target cells.

slide3

IL-2 Gene

+1

+47

-286

-257

-256

-242

-208

-188

-158

-145

-93

-66

TATA

NF-AT

NFIL-2D

NFIL-2C

NFIL-2B

NFIL-2A

NFAT Z Construct

-70

+1

+47

TATA

lac Z

TK

promoter

Hygromycin

Resistance gene

NF-AT

NF-AT

NF-AT

IL-2

slide4

Detection of ß-Galactosidase:

Upon addition of its chromogenic substrate, chlorophenol red galactoside (CPRG), ß-galactosidase hydrolyzes CPRG, turning it from a yellow color to purple at alkaline pH.

The color changs thus provide a convenient measure of gene transcription activities during T cell activation.

slide5

2+

Ca

Receptor

CRAC

2+

Ca

IP3R

Ras, Rac, PKC

PO3

CsA/Cph

NF-ATc

Calcineurin

NF-AT

kinase

NF-ATn

NF-ATc

IL-2

T cell

receptor

activation

Ca2+

NF-ATc

IP3

Phosphoinositol

pathway

IL-2

DAG

PKC

NF-ATn

slide6

2+

Ca

2+

Ca

PO3

NF-ATc

Con A

EGTA

Receptor

CRAC

Ionomycin

thapsigargin

IP3R

Ras, Rac, PKC

CsA/Cph

PMA

Calcineurin

NF-AT

kinase

NF-ATn

NF-ATc

IL-2

slide7

1. Mammalian cell culture:

Goal: learn how to count cells using a hemacytometer

Learn to grow mammalian cell lines

2. Signal transduction lab:

Examine the effects of various inhibitors and activators on

IL-2 expression.

This week’s experiments

slide8

How to count cells using a hemacytometer?

Cell unit per box: 1 x 104 cells/ml

Blue cells : 3 x 104/ml

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