title: Immunogenicity of H-2Kb-low affinity, high affinity, and covalently-bound peptides in anti-tumor vaccination.
abstract: CTL induction by immunization with synthetic peptide epitopes has been shown to inhibit tumor growth and its metastatic spread. Ex vivo pulsing of peptides on MHC class I-bearing cells such as RMA-S cells or professional APCs elicits an effective CTL response. Since the stability of the MHC-peptide complex is strongly correlated with the overall immunogenecity, we compared the effect of immunization with low affinity, high affinity, and irreversibly bound MHC peptides in the context of immunotherapy of metastasis. MUT1, a tumor-associated antigen peptide that was isolated from 3LL Lewis lung carcinoma, is a low H-2Kb binder. MUT1 was modified into a high binder by changing positions 3, 5, and 8 to the favorable anchor residues. In addition, we introduced a photo-active chemical moiety, which can bind irreversibly to MHC upon illumination. These peptides, loaded onto RMA-S, were used to immunize mice against the 3LL tumor. Vaccination via the covalent conjugation of the low binder peptide was found to increase the CTL response measured against MUT1 loaded cells and against H-2Kb transfected D122 cells relative to the native MUT1 peptide. However, the photo cross-linking of the high affinity peptide to the MHC did not significantly improve the induction of specific CTL. The level of CTL activity was elevated to the same extent by either cross-linking the peptide to the MHC or by modifying it into a high-binder peptide. The protective capacity of all the peptide-based vaccines against D122 metastatic spread to the lungs was found to be comparable. These results indicate that augmentation of the affinity of a TAA peptide to the RMA-S surface MHC molecules, by conversion to a high-affinity mimotope or by photo-conjugation, can significantly enhance the immune response. There seems to be, however, a ceiling beyond which increase in the peptide-binding affinity does not lead to a corresponding enhancement of the overall immunogenicity of the peptide.
authors.affiliation: Department of Organic Chemistry, The Weizmann Institute of Science, Rehovot, Israel.
authors.forename: B
authors.initials: B
authors.lastname: Tirosh
authors.forename: K
authors.initials: K
authors.lastname: el-Shami
authors.forename: N
authors.initials: N
authors.lastname: Vaisman
authors.forename: L
authors.initials: L
authors.lastname: Carmon
authors.forename: E
authors.initials: E
authors.lastname: Bar-Haim
authors.forename: E
authors.initials: E
authors.lastname: Vadai
authors.forename: M
authors.initials: M
authors.lastname: Feldman
authors.forename: M
authors.initials: M
authors.lastname: Fridkin
authors.forename: L
authors.initials: L
authors.lastname: Eisenbach
mesh_terms: D000818:Animals; D019496:Cancer Vaccines / Q000737:chemistry / Q000276:immunology*; D017690:Cell Transplantation; D000939:Epitopes / Q000276:immunology; D006183:H-2 Antigens / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018716:Molecular Mimicry; D010455:Peptides / Q000737:chemistry / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014407:Tumor Cells, Cultured; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology*
chemical_list: D019496:Cancer Vaccines; D000939:Epitopes; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D010455:Peptides; D014614:Vaccines, Synthetic