High-throughput identification and dendritic cell-based functional validation of MHC class I-restricted Mycobacterium tuberculosis epitopes.
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.
Duke Scholars
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- Tuberculosis
- T-Lymphocytes, Cytotoxic
- T-Lymphocyte Subsets
- Protein Binding
- Peptides
- Mycobacterium tuberculosis
- Mice, Transgenic
- Mice
- Histocompatibility Antigens Class I
- High-Throughput Screening Assays
Citation
Published In
DOI
EISSN
Publication Date
Volume
Start / End Page
Location
Related Subject Headings
- Tuberculosis
- T-Lymphocytes, Cytotoxic
- T-Lymphocyte Subsets
- Protein Binding
- Peptides
- Mycobacterium tuberculosis
- Mice, Transgenic
- Mice
- Histocompatibility Antigens Class I
- High-Throughput Screening Assays