title: Immune reactivity against a novel HLA-A3-restricted influenza virus peptide identified by predictive algorithms and interferon-gamma quantitative PCR.
abstract: The use of appropriate antigenic peptides for the most common human major histocompatibility complex (MHC) alleles is required for the amplification of the autologous cytotoxic compartment and the development of cytotoxic T cell-mediated immunity. The human A2 allele of the MHC plays an important role for the identification of peptide-specific cytotoxic T cells (CTL) against tumor and viral epitopes. Computer-based prediction algorithms, which are available on the Internet, have already proved to be applicable for the identification of novel CTL epitopes. Using the bioinformatics approach, the authors have identified the novel influenza matrix protein-derived and HLA-A3-restricted 9-mer peptide RLEDVFAGK capable of inducing peptide specific CTL reactivity. Peripheral blood mononuclear cells (PBMC) from healthy individuals and patients with lung cancer were pulsed with this peptide and with the well-characterized HLA-A2-restricted influenza A virus matrix peptide GILGFVFTL. Using quantitative PCR (TaqMan; Applied Biosystems, Foster City, CA, U.S.A), reactivity for both peptides was determined by measuring the change in type 1 cytokine (IFN-gamma) expression upon in vitro stimulation. Peptide-specific reactivity matched well with the subsequently determined MHC-class I alleles of the tested individuals. Results from this study indicate that the use of bioinformatics and the PCR-based screening system for the monitoring of T cell reactivity may allow for the identification of novel CTL epitopes.
authors.affiliation: Division of Oncology, Department of Internal Medicine, University Hospital Zürich, Rämistrasse 100, 8091 Zürich, Switzerland. andreas.trojan@dim.usz.ch
authors.forename: Andreas
authors.initials: A
authors.lastname: Trojan
authors.forename: Mirjana
authors.initials: M
authors.lastname: Urosevic
authors.forename: Jörg
authors.initials: J
authors.lastname: Hummerjohann
authors.forename: Robin
authors.initials: R
authors.lastname: Giger
authors.forename: Urs
authors.initials: U
authors.lastname: Schanz
authors.forename: Rolf A
authors.initials: RA
authors.lastname: Stahel
mesh_terms: D000465:Algorithms; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood; D018984:Epitopes, T-Lymphocyte; D015791:HLA-A3 Antigen / Q000276:immunology*; D006801:Humans; D007114:Immunization; D007371:Interferon-gamma / Q000032:analysis; D008175:Lung Neoplasms / Q000097:blood; D008285:Major Histocompatibility Complex / Q000276:immunology*; D009975:Orthomyxoviridae / Q000276:immunology*; D010446:Peptide Fragments; D016133:Polymerase Chain Reaction / Q000379:methods; D011237:Predictive Value of Tests; D011336:Probability; D012680:Sensitivity and Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D018984:Epitopes, T-Lymphocyte; D015791:HLA-A3 Antigen; D010446:Peptide Fragments; D007371:Interferon-gamma