title: High-affinity interactions between peptides and heat shock protein 70 augment CD8+ T lymphocyte immune responses.
abstract: Exogenously delivered antigenic peptides complexed to heat shock proteins (HSPs) are able to enter the endogenous Ag-processing pathway and prime CD8+ CTL. It was determined previously that a hybrid peptide containing a MHC class I-binding epitope and HSP70-binding sequence Javelin (J0) in complex with HSP70 could induce cytotoxic T cell responses in vivo that were more robust than those induced by the minimal epitope complexed with HSP70. The present study introduces a novel, higher-affinity HSP70-binding sequence (J1) that significantly enhances binding of various antigenic peptides to HSP70. A competition binding assay revealed a dissociation constant that was 15-fold lower for the H2-K(b) OVA epitope SIINFEKL-J1 compared with SIINFEKL-J0, indicating a substantially higher affinity for HSP70. Further, modifying the orientation of the hybrid epitope and introducing a cleavable linker sequence between the Javelin and the epitope results in even greater immunogenicity, presumably by greater efficiency of epitope processing. The enhanced immunogenicity associated with Javelin J1 and the cleavable linker is consistently observed with multiple mouse and human epitopes. Thus, by creating a series of epitopes with uniform, high-affinity binding to HSP70, successful multiple epitope immunizations are possible, with equal delivery of each antigenic epitope to the immune system via HSP70. These modified epitopes have the potential for creating successful multivalent vaccines for immunotherapy of both infectious disease and cancer.
authors.affiliation: Antigenics Inc., 3 Forbes Road, Lexington, MA 02421, USA.
authors.forename: Jessica B
authors.initials: JB
authors.lastname: Flechtner
authors.forename: Kenya Prince
authors.initials: KP
authors.lastname: Cohane
authors.forename: Sunil
authors.initials: S
authors.lastname: Mehta
authors.forename: Paul
authors.initials: P
authors.lastname: Slusarewicz
authors.forename: Alexis Kays
authors.initials: AK
authors.lastname: Leonard
authors.forename: Brian H
authors.initials: BH
authors.lastname: Barber
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Levey
authors.forename: Sofija
authors.initials: S
authors.lastname: Andjelic
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D018414:CD8-Positive T-Lymphocytes / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000276:immunology; D002417:Cattle; D045744:Cell Line, Tumor; D045142:Cross-Priming; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000502:physiology; D011485:Protein Binding / Q000276:immunology; D025941:Protein Interaction Mapping; D013945:Thymoma / Q000276:immunology / Q000473:pathology / Q000517:prevention & control
chemical_list: D000276:Adjuvants, Immunologic; D019496:Cancer Vaccines; D018984:Epitopes, T-Lymphocyte; D018840:HSP70 Heat-Shock Proteins; D010446:Peptide Fragments; D010047:Ovalbumin; D046988:Proteasome Endopeptidase Complex