title: Efficient induction of cytotoxic CD8+ T cells against exogenous proteins: establishment and characterization of a T cell line specific for the membrane protein ActA of Listeria monocytogenes.
abstract: The property of listeriolysin (LLO) to introduce soluble passenger proteins into the cytosol of antigen-presenting cells allows the induction of CD8+ cytotoxic T cells against such antigens. To overcome the potential problem of presentation of the immunodominant epitope LL091-99 by H-2Kd, a variant LLO92A was established in which Tyr 92 was replaced by Ala. Immunization of BALB/c mice with purified LLO92A failed to stimulate cytotoxic T cells specific for either the epitope LLO91-99 or for any other LLO-derived peptide. Injection of mixtures of purified LLO92A and soluble nucleoprotein (NP) of influenza virus into mice resulted in a strong cytotoxic T cell response exclusively directed against NP. The LLO92A variant was successfully used to generate, propagate and characterize a CD8 T cell line specific for the membrane-bound virulence factor ActA of Listeria monocytogenes. Interestingly, wildtype ActA bound to the surface of live L. monocytogenes was not presented by MHC class I molecules to the CD8+ T cell line.
authors.affiliation: Department of Cell Biology and Immunology, GBF, National Research Center for Biotechnology, Braunschweig, Germany. dbr@gbf.de
authors.forename: D
authors.initials: D
authors.lastname: Bruder
authors.forename: A
authors.initials: A
authors.lastname: Darji
authors.forename: D M
authors.initials: DM
authors.lastname: Gakamsky
authors.forename: T
authors.initials: T
authors.lastname: Chakraborty
authors.forename: I
authors.initials: I
authors.lastname: Pecht
authors.forename: J
authors.initials: J
authors.lastname: Wehland
authors.forename: S
authors.initials: S
authors.lastname: Weiss
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D000942:Antigens, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology*; D001427:Bacterial Toxins; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology; D006460:Hemolysin Proteins; D008089:Listeria monocytogenes / Q000276:immunology*; D008565:Membrane Proteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009154:Mutation
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D008565:Membrane Proteins; C072980:actA protein, Listeria monocytogenes; C038705:hlyA protein, Listeria monocytogenes