title: Detection of HLA class II-dependent T helper antigen using antigen phage display.
abstract: Major histocompatibility complex (MHC) class II-dependent antigens not only activate CD4+ T helper (Th) cells, but also cytolytic T lymphocytes and effector cells of the innate immune system. These antigens therefore are candidate vaccines against cancer and infectious agents. We have developed a novel approach using a model antigen, tetanus toxoid (TT), which provides the basis for the establishment of a novel strategy of cloning Th antigens. In the TT model system, a cDNA library encoding part of the TT light chain which contained a TT-associated Th epitope recognized by TT-specific Th clones was displayed on a phage vector (TT-phage) and presented to TT-specific Th cells by autologous Epstein-Barr virus-transformed B cells (APC). These TT-phages were able to specifically activate two different TT-specific CD4+ Th cell lines as demonstrated both in [3H]thymidine incorporation and cytokine release assays. Th cell stimulation by TT-phages was significant at a ratio of one TT-phage in 50 irrelevant phages. The described approach provides the basis for the development of a novel strategy of cloning MHC class II-dependent Th antigens, using available Th cells. This strategy has several potential advantages over existing antigen cloning methods or biochemical peptide isolation.
authors.affiliation: The Wistar Institute, Philadelphia, PA 19104, USA.
authors.forename: R
authors.initials: R
authors.lastname: Somasundaram
authors.forename: K
authors.initials: K
authors.lastname: Satyamoorthy
authors.forename: L
authors.initials: L
authors.lastname: Caputo
authors.forename: H
authors.initials: H
authors.lastname: Yssel
authors.forename: D
authors.initials: D
authors.lastname: Herlyn
mesh_terms: D017951:Antigen Presentation / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D001435:Bacteriophages / Q000276:immunology*; D015704:CD4 Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D016207:Cytokines / Q000032:analysis; D004279:DNA, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D006801:Humans; D013745:Tetanus Toxoid / Q000276:immunology*; D013936:Thymidine / Q000276:immunology; D014764:Viral Proteins / Q000032:analysis
chemical_list: D015704:CD4 Antigens; D016207:Cytokines; D004279:DNA, Viral; D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D013745:Tetanus Toxoid; D014764:Viral Proteins; D013936:Thymidine