Sequence arrangements and sequences for neoepitope presentation
Systems and methods are presented that allow for selection of tumor neoepitopes that are then used to generate a recombinant polytope that is optimized for proper trafficking and processing. In preferred methods, the polytope is encoded in a viral expression system that is used as a therapeutic agent.
1. A method of eliciting an immune response against a tumor in a subject, the method comprising:
determining a common sequence location within first and second sequence datasets, wherein the first sequence dataset comprises sequence data from a tumor in the subject, and wherein the second sequence dataset comprises sequence data from a matched normal tissue in the subject;
generating at least one pair of sequence pileups from the first and second sequence datasets that overlap at the common sequence location;
determining genotypes for healthy tissue and suspected diseased tissue at the common sequence location as a function of the at least one pair of sequence pileups;
identifying a sequence difference at the common sequence location and between genotypes;
filtering out false positives based on reads in the at least one pair of sequence pileups to thus generate at least one patient specific neoepitope;
immunizing the subject in need thereof with IL-15, RP-182, and a recombinant adenovirus;
wherein the recombinant adenovirus comprises a nucleic acid sequence encoding the at least one patient specific neoepitopes operably linked to a trafficking element that directs the nucleic acid sequence to a sub-cellular location selected from the group consisting of recycling endosome, sorting endosome, and lysosome.
2. The method of claim 1 , wherein the at least one patient specific neoepitope is filtered for MHC binding of less than or equal to 200 nM.
3. The method of claim 1 , wherein the recombinant nucleic acid further comprises a sequence of at least one additional neoepitope, wherein the additional neoepitope is operably linked to a trafficking element that directs the additional neoepitope to a different sub-cellular location than the at least one patient specific neoepitope.
4. The method of claim 1 , wherein the recombinant adenovirus further comprises a sequence encoding a protein selected from the group consisting of CD80, CD86, CD30, CD40, CD30L, CD40L, ICOS-L, B7-H3, B7-H4, CD70, OX40L, 4-1BBL, GITR-L, TIM-3, TIM-4, CD48, CD58, TL1A, ICAM-1, LFA3, IL-2, IL-12, IL-15, IL-15 super agonist (ALT803), IL-21, IPS1, LMP1, an anti-CTLA-4 antibody, an anti-PD-1 antibody, an anti-TIM1 receptor antibody, an anti-2B4 antibody, and an anti-CD160 antibody.