title: High-throughput identification and dendritic cell-based functional validation of MHC class I-restricted Mycobacterium tuberculosis epitopes.
abstract: Emergence of drug-resistant strains of the pathogen Mycobacterium tuberculosis (Mtb) and the ineffectiveness of BCG in curtailing Mtb infection makes vaccine development for tuberculosis an important objective. Identifying immunogenic CD8+ T cell peptide epitopes is necessary for peptide-based vaccine strategies. We present a three-tiered strategy for identifying and validating immunogenic peptides: first, identify peptides that form stable complexes with class I MHC molecules; second, determine whether cytotoxic T lymphocytes (CTLs) raised against the whole protein antigen recognize and lyse target cells pulsed with peptides that passed step 1; third, determine whether peptides that passed step 2, when administered in vivo as a vaccine in HLA-A2 transgenic mice, elicit CTLs that lyse target cells expressing the whole protein antigen. Our innovative approach uses dendritic cells transfected with Mtb antigen-encoding mRNA to drive antigen expression. Using this strategy, we have identified five novel peptide epitopes from the Mtb proteins Apa, Mtb8.4 and Mtb19.
authors.affiliation: 1] Departments of Surgery, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Smita K
authors.initials: SK
authors.lastname: Nair
authors.affiliation: 1] Departments of Surgery, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Georgia D
authors.initials: GD
authors.lastname: Tomaras
authors.affiliation: 1] Biostatistics and Bioinformatics, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Sales
authors.affiliation: Departments of Surgery, Duke University Medical Center, Durham, NC 27710.
authors.forename: David
authors.initials: D
authors.lastname: Boczkowski
authors.affiliation: Biostatistics and Bioinformatics, Duke University Medical Center, Durham, NC 27710.
authors.forename: Cliburn
authors.initials: C
authors.lastname: Chan
authors.affiliation: Departments of Surgery, Duke University Medical Center, Durham, NC 27710.
authors.forename: Kelly
authors.initials: K
authors.lastname: Plonk
authors.affiliation: 1] Departments of Surgery, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Yongting
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Departments of Surgery, Duke University Medical Center, Durham, NC 27710.
authors.forename: Jens
authors.initials: J
authors.lastname: Dannull
authors.affiliation: 1] Biostatistics and Bioinformatics, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Kepler
authors.affiliation: 1] Departments of Surgery, Duke University Medical Center, Durham, NC 27710 [2].
authors.forename: Scott K
authors.initials: SK
authors.lastname: Pruitt
authors.affiliation: Departments of Surgery, Duke University Medical Center, Durham, NC 27710.
authors.forename: Kent J
authors.initials: KJ
authors.lastname: Weinhold
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism; D018604:Epitope Mapping / Q000379:methods*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D057166:High-Throughput Screening Assays; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology*; D010455:Peptides / Q000276:immunology; D011485:Protein Binding / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014376:Tuberculosis / Q000276:immunology*
chemical_list: D000942:Antigens, Bacterial; D018984:Epitopes, T-Lymphocyte; D015395:Histocompatibility Antigens Class I; D010455:Peptides
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