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.docVIP

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 1,3* Christine Almunia , Marie Bretaudeau 3 2 1 3 , Gerhard Held , Aurélie Babon , Charles Marchetti , Florence 1 Anne Castelli , André Ménez , Bernard Maillere , Daniel Gillet 1,4 1 1 1 Service d’Ingénierie Moléculaire des Protéines, Institut de Biologie et Technologies de Saclay, Commissariat à lénergie atomique et aux énergies alternatives, Gif Sur Yvette, France, 2 Medizinische Klinik I, Universitaetsklinik des Saarlandes, Homburg, Germany, 3 Service de Biochimie et de Toxicologie nucléaire, Institut de Biologie Environnementale et Biotechnologie, Commissariat à lénergie atomique et aux énergies alternatives, Bagnols sur Cèze, France, 4 Museum National dHistoire Naturelle, Paris, France 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