title: Cross-presentation of HLA class I epitopes from influenza matrix protein produced in Saccharomyces cerevisiae.
abstract: Here we report that genetically engineered yeast of the strain Saccharomyces cerevisiae expressing full-length influenza matrix protein (IMP) attached to the yeast cell wall are a very versatile host for antigen delivery. Feeding of dendritic cells with either intact yeast expressing IMP protein or soluble IMP protein cleaved off the cell wall resulted in protein uptake, processing and cross-presentation of IMP-derived peptides. This process was analysed using previously established T-cell lines recognizing the immuno-dominant 58-66 peptide when presented by HLA-A2*0201 complexes. In addition, IMP(58-66)/HLA-A2*0201-specific antibodies were selected from a naive phage library which confirmed that peptide presentation was an active process of endocellular uptake and not just a result of external peptide loading. Moreover, MHC peptide antibodies could block the recognition of peptide-presenting dendritic cells by IMP(58-66)-specific T-cells in a dose dependent manner. There was no difference in T-cell recognition when either intact yeast or yeast cell extracts were used for DC feeding. Together, these data demonstrate that yeast derived proteins either in their soluble form or as part of a whole yeast vaccine are taken up, processed and presented by dendritic cells in HLA class I context.
authors.affiliation: Klinik und Poliklinik für Onkologie, Rämistr. 100, UniversitätsSpital Zürich, CH-8091 Zürich, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Wadle
authors.forename: Gerhard
authors.initials: G
authors.lastname: Held
authors.forename: Frank
authors.initials: F
authors.lastname: Neumann
authors.forename: Sascha
authors.initials: S
authors.lastname: Kleber
authors.forename: Beate
authors.initials: B
authors.lastname: Wuellner
authors.forename: Anne Marie
authors.initials: AM
authors.lastname: Asemissen
authors.forename: Boris
authors.initials: B
authors.lastname: Kubuschok
authors.forename: Carmen
authors.initials: C
authors.lastname: Scheibenbogen
authors.forename: Tanja
authors.initials: T
authors.lastname: Breinig
authors.forename: Andreas
authors.initials: A
authors.lastname: Meyerhans
authors.forename: Christoph
authors.initials: C
authors.lastname: Renner
mesh_terms: D000918:Antibody Specificity; D017951:Antigen Presentation / Q000276:immunology; D051940:B7-2 Antigen / Q000276:immunology; D045142:Cross-Priming / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology; D004705:Endocytosis / Q000276:immunology; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology*; D015789:HLA-A2 Antigen; D016056:Immunodominant Epitopes / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D037181:Lectins, C-Type / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins / Q000276:immunology; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology*; D010587:Phagocytosis / Q000276:immunology; D011956:Receptors, Cell Surface / Q000276:immunology; D015375:Receptors, Interleukin-2 / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000276:immunology*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D051940:B7-2 Antigen; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D016056:Immunodominant Epitopes; D007128:Immunoglobulin Fragments; D037181:Lectins, C-Type; D018950:Lipopolysaccharide Receptors; D000090323:Mannose Receptor; D037241:Mannose-Binding Lectins; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014763:Viral Matrix Proteins; C448634:influenza matrix peptide (58-66); C069939:a-agglutinin protein, S cerevisiae