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.
High-throughput identification and dendritic cell-based functional validation of MHC class I-restricted Mycobacterium tuberculosis epitopes
S. Nair,G. Tomaras,A. Sales,D. Boczkowski,Cliburn Chan,K. Plonk,Yongting Cai,J. Dannull,T. Kepler,S. Pruitt,K. Weinhold
Published 2014 in Scientific Reports
ABSTRACT
PUBLICATION RECORD
- Publication year
2014
- Venue
Scientific Reports
- Publication date
2014-04-23
- Fields of study
Biology, Medicine
- Identifiers
- External record
- Source metadata
Semantic Scholar, PubMed
CITATION MAP
EXTRACTION MAP
CLAIMS
CONCEPTS
- apa
A Mycobacterium tuberculosis protein included as a source of identified peptide epitopes in this study.
Aliases: Mtb Apa
- dendritic cells transfected with antigen-encoding mrna
Dendritic cells engineered with mRNA so they express the encoded Mtb antigen and present derived peptides for immune testing.
Aliases: dendritic cells transfected with Mtb antigen-encoding mRNA
- five novel peptide epitopes
The set of newly identified Mtb-derived peptide epitopes reported as validated candidates.
Aliases: five novel epitopes
- hla-a2 transgenic mice
A mouse model engineered to express human HLA-A2 used for in vivo vaccine and CTL validation.
Aliases: HLA-A2 mice, transgenic mice
- mtb19
A Mycobacterium tuberculosis protein included as a source of identified peptide epitopes in this study.
Aliases: Mtb 19
- mtb8.4
A Mycobacterium tuberculosis protein included as a source of identified peptide epitopes in this study.
Aliases: Mtb 8.4
- mtb antigen-encoding mrna
Messenger RNA constructs that encode Mycobacterium tuberculosis antigens for expression in dendritic cells.
Aliases: antigen-encoding mRNA
- stable class i mhc complexes
Peptide-class I MHC complexes that remain sufficiently stable to pass the initial screening step for candidate epitopes.
Aliases: stable complexes with class I MHC molecules, MHC class I binding stability
- three-tiered epitope validation strategy
A sequential workflow that screens peptide candidates by MHC class I binding, CTL recognition of peptide-pulsed targets, and in vivo functional validation.
Aliases: three-tiered strategy, three-tiered strategy for identifying and validating immunogenic peptides
REFERENCES
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