T cell protocols : development and activation

pt. 1. Overview: for chapters 2-15 -- Insights into T-cell development using transgenic and knockout mice -- pt. 2. T cell development -- Isolation and characterizatin of murine early intrathymic precursor populations -- Differentiation of mouse thymocytes in fetal thymus organ culture -- Purification of immature CD4+CD8+ thymyocytes by panning with anti-CD8 antibody -- Transfection and transcription of genes in developing thymocytes -- analysis of CD4/CD8 lineage commitment by pronase treatment and reexpression assay -- Intrahymic delivery of MHC genes using recombinant adenoviruses -- Quantitative analysis of the usage of human T-cell receptor [alpha] and [beta] chain variable regions by reverse dot-blot hybridization -- Flow cytometric detection of cytoplasmic and surface CD3-[epsilon] expression in developing T cells -- Assessing apoptosis in developing T cells by flow cytometry -- Isolation of T-cells subsets by magnetic cell sorting (MACS) -- Isolation of T-cell antigens by retrovirus-mediated expression cloning -- Flow cytometric analysis of murine T lymphocytes: a practical guide -- Production and characterization of T cell hybridomas -- pt. 3. Overview: for chapters 17-29 --Something happens: a brief history of TCR signal transduction -- pt. 4. T-cell activation -- Measurement of phosphatidylinositol (PI) hydrolysis in activated T lymphocytes -- Fluorescence polarization as an early measure of T-lymphocyte stimulation -- Measurement of lymphoproliferation at the single-cell level by flow cytometry -- Digital image analysis of lymphocyte activation -- Induction of T-cell signaling by immobilized integrin ligands -- Cytotoxic T-cell adherence assay (CAA) -- Detecting ubiquitinated T-cell antigen receptor subunits by immunoblotting -- Determination of CD45 tyrosine phosphatase activity in T lymphocytes -- Measurement of protein tyrosine phosphorylation in T-cell subsets by flow cytometry -- Biochemical analysis of activated T lymphocytes: protein phosphorylation and Ras, ERK and JNK activation -- Activation of heterotrimeric GTP-binding proteins upon TCR/CD3 engagement -- Generation of human T helper 1 and T helper 2 subsets form peripheral blood-derived naive CD4+ T cells -- Transcriptional expression analysis of T-cell activation by multiplex meassenger assay (MMA).