title: Bee Venom Phospholipase A2, a Good "Chauffeur" for Delivering Tumor Antigen to the MHC I and MHC II Peptide-Loading Compartments of the Dendritic Cells: The Case of NY-ESO-1.
abstract: Bee venom phospholipase A2 (bvPLA2) is a small, 15kDa enzyme which hydrolyses many phospholipids through interfacial binding. The mutated bvPLA2H34Q (bvPLA2m), in which histidine-34 is replaced by glutamine, is not catalytically active. This protein has been shown to be a suitable membrane anchor and has been suggested as a suitable tumor-antigen vector for the development of novel dendritic cell-based vaccines. To confirm this feature, in this study the fusion protein PNY, composed of NY-ESO-1(NY(s)) fused to the C-terminus of bvPLA2m, was engineered. bvPLA2m enhanced the binding of NY(s) to the membrane of human monocyte-derived dendritic cells (DCs) and, once taken up by the cells, the antigen fused to the vector was directed to both MHC I and MHC II peptide-loading compartments. bvPLA2m was shown to increase the cross-presentation of the NY(s)-derived, restricted HLA-A*02 peptide, NY-ESO-1157-165(NY157-165), at the T1 cell surface. DCs loaded with the fusion protein induced cross-priming of NY(s)-specific CD8 + T-cells with greater efficiency than DCs loaded with NY(s). Sixty-five percent of these NY(s)-specific CD8+ T-cell lines could also be activated with the DCs pulsed with the peptide, NY157-165. Of these CD8+ T-cell lines, two were able to recognize the human melanoma cell line, SK-MEL-37, in a context of HLA-A*02. Only a small number of bvPLA2m CD8+ T-cell lines were induced, indicating the low immunogenicity of the protein. It was concluded that bvPLA2m can be used as a membrane-binding vector to promote MHC class II peptide presentation and MHC class I peptide cross-presentation. Such a system can, therefore, be tested for the preparation of cell-based vaccines.
authors.affiliation: Service d'Ingénierie Moléculaire des Protéines, Institut de Biologie et Technologies de Saclay, Commissariat à l'énergie atomique et aux énergies alternatives, Gif Sur Yvette, France ; Service de Biochimie et de Toxicologie nucléaire, Institut de Biologie Environnementale et Biotechnologie, Commissariat à l'énergie atomique et aux énergies alternatives, Bagnols sur Cèze, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Almunia
authors.forename: Marie
authors.initials: M
authors.lastname: Bretaudeau
authors.forename: Gerhard
authors.initials: G
authors.lastname: Held
authors.forename: Aurélie
authors.initials: A
authors.lastname: Babon
authors.forename: Charles
authors.initials: C
authors.lastname: Marchetti
authors.forename: Florence Anne
authors.initials: FA
authors.lastname: Castelli
authors.forename: André
authors.initials: A
authors.lastname: Ménez
authors.forename: Bernard
authors.initials: B
authors.lastname: Maillere
authors.forename: Daniel
authors.initials: D
authors.lastname: Gillet
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism*; D001514:Bee Venoms / Q000201:enzymology*; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D008285:Major Histocompatibility Complex / Q000276:immunology*; D054467:Phospholipases A2 / Q000378:metabolism*
chemical_list: D000951:Antigens, Neoplasm; D001514:Bee Venoms; D054467:Phospholipases A2