title: Isolation of neoantigen-specific T cells from tumor and peripheral lymphocytes.
abstract: Adoptively transferred tumor-infiltrating T lymphocytes (TILs) that mediate complete regression of metastatic melanoma have been shown to recognize mutated epitopes expressed by autologous tumors. Here, in an attempt to develop a strategy for facilitating the isolation, expansion, and study of mutated antigen-specific T cells, we performed whole-exome sequencing on matched tumor and normal DNA isolated from 8 patients with metastatic melanoma. Candidate mutated epitopes were identified using a peptide-MHC-binding algorithm, and these epitopes were synthesized and used to generate panels of MHC tetramers that were evaluated for binding to tumor digests and cultured TILs used for the treatment of patients. This strategy resulted in the identification of 9 mutated epitopes from 5 of the 8 patients tested. Cells reactive with 8 of the 9 epitopes could be isolated from autologous peripheral blood, where they were detected at frequencies that were estimated to range between 0.4% and 0.002%. To the best of our knowledge, this represents the first demonstration of the successful isolation of mutation-reactive T cells from patients' peripheral blood prior to immune therapy, potentially providing the basis for designing personalized immunotherapies to treat patients with advanced cancer.
authors.forename: Cyrille J
authors.initials: CJ
authors.lastname: Cohen
authors.forename: Jared J
authors.initials: JJ
authors.lastname: Gartner
authors.forename: Miryam
authors.initials: M
authors.lastname: Horovitz-Fried
authors.forename: Katerina
authors.initials: K
authors.lastname: Shamalov
authors.forename: Kasia
authors.initials: K
authors.lastname: Trebska-McGowan
authors.forename: Valery V
authors.initials: VV
authors.lastname: Bliskovsky
authors.forename: Maria R
authors.initials: MR
authors.lastname: Parkhurst
authors.forename: Chen
authors.initials: C
authors.lastname: Ankri
authors.forename: Todd D
authors.initials: TD
authors.lastname: Prickett
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Crystal
authors.forename: Yong F
authors.initials: YF
authors.lastname: Li
authors.forename: Mona
authors.initials: M
authors.lastname: El-Gamil
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rosenberg
authors.forename: Paul F
authors.initials: PF
authors.lastname: Robbins
mesh_terms: D000293:Adolescent; D000328:Adult; D000465:Algorithms; D000595:Amino Acid Sequence; D000937:Antigen-Antibody Reactions; D000951:Antigens, Neoplasm / Q000145:classification / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D004273:DNA, Neoplasm / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D059472:Exome; D005260:Female; D018734:Genes, erbB-2; D015788:HLA-A1 Antigen / Q000737:chemistry / Q000276:immunology; D015789:HLA-A2 Antigen / Q000737:chemistry / Q000276:immunology; D006801:Humans; D059425:Interferon-gamma Release Tests; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008297:Male; D008545:Melanoma / Q000235:genetics / Q000276:immunology* / Q000556:secondary*; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology; D009687:Nuclear Proteins / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D012334:RNA, Neoplasm / Q000235:genetics*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology; D037182:T-Cell Antigen Receptor Specificity; D013601:T-Lymphocytes / Q000276:immunology*; D000091262:TEA Domain Transcription Factors; D014157:Transcription Factors / Q000235:genetics / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D000939:Epitopes; D015788:HLA-A1 Antigen; D015789:HLA-A2 Antigen; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D012334:RNA, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases; D000091262:TEA Domain Transcription Factors; D014157:Transcription Factors; C435939:HLA-A*02:01 antigen; C057182:TEAD1 protein, human; C508053:ERBB2 protein, human