title: Signal peptides and trans-membrane regions are broadly immunogenic and have high CD8+ T cell epitope densities: Implications for vaccine development.
abstract: Cell mediated immune response has a major role in controlling the elimination of infectious agents. The rational design of sub-unit peptide vaccines against intracellular pathogens or cancer requires the use of antigenic sequence/s that can induce highly potent, long lasting and antigen-specific responses in the majority of the population. A promising peptide selection strategy is the detection of multi-epitope peptide sequences with an ability to bind multiple MHC alleles. While past research sought the best epitopes based on their specific antigenicity, we ask whether specific defined domains have high epitope densities. Signal peptides and trans-membrane domains were found to have exceptionally high epitope densities. The improved MHC binding of these domains relies on their hydrophobic nature and, in signal peptides, also on their specific sequence. The high epitope density of SP was computed using in-silico methods and corroborated by the high percentage of identified SP epitope in the IEDB (immune epitope database). The enhanced immunogenicity of SP was then experimentally confirmed using a panel of nine peptides derived from Mycobacterium tuberculosis (MTb) proteins used in human PBMC proliferation assays and T cell lines functional assays. Our results show the exceptionally high antigen specific response rates and population coverage to SP sequences compared with non-SP peptide antigens derived from the same proteins. The results suggest a novel scheme for the rational design of T cell vaccines using a domain based rather than an epitope based approach.
authors.affiliation: Vaxil BioTherapeutics Ltd. 13A, WIS Science Park, Nes-Ziona 74036, Israel.
authors.forename: Riva
authors.initials: R
authors.lastname: Kovjazin
authors.forename: Ilan
authors.initials: I
authors.lastname: Volovitz
authors.forename: Yair
authors.initials: Y
authors.lastname: Daon
authors.forename: Tal
authors.initials: T
authors.lastname: Vider-Shalit
authors.forename: Roy
authors.initials: R
authors.lastname: Azran
authors.forename: Lea
authors.initials: L
authors.lastname: Tsaban
authors.forename: Lior
authors.initials: L
authors.lastname: Carmon
authors.forename: Yoram
authors.initials: Y
authors.lastname: Louzoun
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D005434:Flow Cytometry; D006649:Histocompatibility Antigens / Q000276:immunology; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000276:immunology; D010455:Peptides / Q000276:immunology / Q000494:pharmacology; D011485:Protein Binding / Q000276:immunology; D021382:Protein Sorting Signals; D022223:Vaccines, Subunit / Q000235:genetics / Q000276:immunology*
chemical_list: D001426:Bacterial Proteins; D018984:Epitopes, T-Lymphocyte; D006649:Histocompatibility Antigens; D008565:Membrane Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D022223:Vaccines, Subunit