[医学]浙大高级免疫学技术英文课件.ppt

An Example Antibodies FACS (Analysis and sorting) ELISA ELISPOT Tetramer cell culture (cells and in vitro culture system) Gene Profiling Proteomics Transgenic and knockout animals Technology based on Immunology Technology which boost immunology MHC tetramers can be used to quantitate numbers of antigen-specific T cells (especially CD8+ T cells). Biotinylated recombinant class I molecules folded with the peptide of interest and β2M and tetramerized by a fluorescently labeled streptavidin(Streptavidin binds to four biotins per molecule.) . This tetramer reagent will specifically label T cells that express T cell receptors that are specific for a given peptide-MHC complex. Antigen specific responses can be measured as CD8+, tetramer+ T cells as a fraction of all CD8+ lymphocytes. Tetramer Tetramers can bind to three TCRs at once, allowing specific binding in spite of the low (10-6 molar) affinity of the typical class I-peptide-TCR interaction. Tetramer CD8 response related(now CD4 too) and known epitope Infectious Disease HIV-AIDS, EBV, CMV, HPV, HBV, HCV, Influenza, Measles, Malaria, TB and RSV Breast, Prostate, Melanoma, Colon, Lung, Cervical, Ovarian and Leukemia Diabetes, Lyme disease, Multiple sclerosis, Rheumatoid arthritis, Autoimmune vitiligo EBV and CMV Transplantation Autoimmune Disease Tumor Immunology Utility of Tetramer Antibodies FACS (Analysis and sorting) ELISA ELISPOT Tetramer cell culture (cells and in vitro culture system) Gene Profiling Proteomics Transgenic and knockout animals Technology based on Immunology Technology which boost immunology The Biggest cell culture resource : American Type Culture Collection (ATCC) Advantage of Cell culture: in vitro study system fast cost less, easy to manipulate Problems in cell culture: 2D vs 3D microenvironment cells mutated during the culture Cell line vs primary cell culture Cell culture Primary cell cul