Classical antigen presentation by HLA-A2 of Mycobacterium tuberculosis derived ligands (106.40)

Abstract Mycobacterium tuberculosis (Mtb) remains a world health threat, with 8.8 million cases reported in 2010, 65% of which were new and recurrent cases of tuberculosis . Cellular immunity is critical for controlling Mtb and recent studies identify CD8+ T cells as important players in controlling infection. Cytotoxic T lymphocytes (CTLs) recognize peptide antigen presented by Major Histocompatibility Complex (MHC) Class I molecules and destroy infected cells, therefore discovery of CTL epitopes is of high importance. Using a method of soluble HLA production, we were able to generate soluble HLA-A2 from THP-1 monocyte cell lines transfected with a soluble form of HLA A*02:01. Transfected cells were seeded into a hollow fiber bioreactor and were infected with Mtb or left uninfected. Following production of 25 mgs of HLA-A2 from infected and uninfected cells, peptides were eluted from the supernatants of uninfected and infected cells. Peptides from each group were fractionated using HPLC and analyzed by mass spectrometry. We were able to identify six Mtb derived ligands presented by the MHC Class I molecule HLA-A*0201, two of which reacted well with donor PBMC from patients with both active and latent Mtb infection. Identifying new vaccine candidates is a top priority in the TB field and the reactive Mtb ligands identified here demonstrate that multiple Mtb proteins are processed for presentation by Class I HLA and a fraction of these ligands are immunologically reactive.