In the course of this joint venture, EpiVax proposes to use its bioinformatics technology, EpiMer and EpiMatrix, to scan the genome of a clinical isolate of Mtb for a broadly immunogenic or promiscuous regions that will be used to develop a epitope-driven oligonucleotide (DNA) vaccine for tuberculosis (TB). We have selected the TIGR TB genome (the """"""""Oshkosh"""""""" strain of Mtb, or CDC stain 1552) as the focus of this TB vaccine development project for two reasons: (I) the entire genome sequence will be available to EpiVax, Inc. when it is completed and (2) a number of open reading frames representing putative proteins that are unique to this pathogenic strain of Mtb will be identified in the process of sequencing that genome. Epitopes from these proteins of interest will be selected according to the following criteria: (l) they will be contained in an QRF unique to Mtb; (2) they will match to a matrix pattern suggesting MHC binding potential for MHC class l and/or class Il molecules; and (3) they will contain a clustering of MHC binding potential (promiscuous epitopes). These putative epitopes will be synthesized and then screened by measuring binding to MHC in vitro and recognition by human T cells. Epitopes that bind to a number of different MHC with high affinity; and/or stimulate T cell responses in assays using cells from genetically diverse individuals will be inserted in string-of-beads DNA constructs; these constructs will be evaluated for successful expression in human cell lines and for immunogenicity in """"""""in vitro vaccination"""""""" studies and in the murine model. EpiVax will then explore, with the assistance of Sequella Global TB Foundation, the efficacy of selected constructs in the non-human primate model of tuberculosis.
