title: Construction and screening of an antigen-derived peptide library displayed on yeast cell surface for CD4+ T cell epitope identification.
abstract: Identification of T cell epitopes is a critical, but often difficult step in studying T cell function and developing peptide-based vaccines and immunotherapies. Unlike antibodies that recognize free soluble antigens, T cell receptor (TCR) recognizes its epitope bound to major histocompatibility complex (MHC) expressed on antigen presenting cells (APCs). In addition, the examination of T cell epitope activity requires the use of professional APCs, which are difficult to isolate, expand, and maintain. To address these issues, we have developed a facile, accurate, and high-throughput method for T cell epitope mapping by screening antigen-derived peptide libraries in complex with MHC protein displayed on yeast cell surface. Here, we use hemagglutinin and influenza A virus X31/A/Aichi/68 as examples to describe the key steps in identification of CD4+ T cell epitopes from a single antigenic protein and the entire genome of a pathogen, respectively. Methods for single-chain peptide-MHC complex vector design, yeast surface display, peptide library generation in Escherichia coli, and functional screening in Saccharomyces cerevisiae are discussed.
authors: Department of Chemical Engineering, University of Michigan, Ann Arbor, MI, USA.
authors: Fei
authors: F
authors: Wen
authors: Huimin
authors: H
authors: Zhao
mesh_terms: D000938:Antigen-Presenting Cells / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D018984:Epitopes, T-Lymphocyte / Q000378:metabolism*; D008285:Major Histocompatibility Complex / Q000502:physiology; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: D018984:Epitopes, T-Lymphocyte; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell