No abstract

Identification of CD8+ T cell antigens/epitopes expressed by human pathogens with large genomes is especially challenging, yet necessary for vaccine development. Immunity to tuberculosis, a leading cause of mortality worldwide, requires CD8+ T cell immunity, yet the repertoire of CD8 antigens/epitopes remains undefined. We used integrated computational and proteomic approaches to screen 10% of the Mycobacterium tuberculosis (Mtb) proteome for CD8 Mtb antigens. We designed a weighting schema based upon a Multiple Attribute Decision Making:framework to select 10% of the Mtb proteome with a high probability of containing CD8+ T cell epitopes. We created a synthetic peptide library consisting of 15-mers overlapping by 11 aa. Using the interferon-γ ELISPOT assay and Mtb-infected dendritic cells as antigen presenting cells, we screened Mtb-specific CD8+ T cell clones restricted by classical MHC class I molecules (MHC class Ia molecules), that were isolated from Mtb-infected humans, against this library. Three novel CD8 antigens were unambiguously identified: the EsxJ family (Rv1038c, Rv1197, Rv3620c, Rv2347c, Rv1792), PE9 (Rv1088), and PE_PGRS42 (Rv2487c). The epitopes are B5701-restricted EsxJ24–34, B3905-restricted PE953–67, and B3514-restricted PE_PGRS4248–56, respectively. The utility of peptide libraries in identifying unknown epitopes recognized by classically restricted CD8+ T cells was confirmed, which can be applied to other intracellular pathogens with large size genomes. In addition, we identified three novel Mtb epitopes/antigens that may be evaluated for inclusion in vaccines and/or diagnostics for tuberculosis.

Mycobacterium tuberculosis (Mtb) remains a major threat to human health worldwide. Although treatment of infection is an important part of tuberculosis control, an improved vaccine is essential for the elimination of this disease. Control of infection with Mtb is dependent on the cellular immune system, which in turn requires an understanding of those antigens that are capable of stimulating CD4+ and CD8+ T-cell responses. Peptide libraries provide a high-throughput system for identifying novel T-cell epitopes. They can also be used to assess the hierarchy of immunodominance of these novel antigens and epitopes that are associated with infection with Mtb. This T-cell-driven means of antigen discovery is well adapted to vaccine development as well as developing the tools necessary to understand the natural history of this important human pathogen.