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  • Poster presentation
  • Open Access

Study and characterization of mutated antigen specific T cells isolated from fresh tumor and peripheral lymphocytes in cancer patients

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Journal for ImmunoTherapy of Cancer20153 (Suppl 2) :P7

  • Published:


  • Melanoma
  • Advanced Cancer
  • Metastatic Melanoma
  • Melanoma Patient
  • Exome Sequencing


T cell-based immunotherapy shows promise for the successful treatment of advanced cancer. Indeed, adoptively transferred tumor infiltrating T lymphocytes (TIL) that mediated complete regression of metastatic melanoma have been shown to recognize neoantigens/mutated epitopes expressed by autologous tumors.


In the present study, we sought to develop a strategy for facilitating the isolation, expansion and study of T cells specific for neoantigens. We performed whole exome sequencing on matched tumor and normal DNA from eight metastatic melanoma patients. Candidate neo-epitopes, identified using a peptide/MHC binding algorithm, were synthesized and we used those to produce panels of MHC/tetramers that were evaluated for binding to tumor digests and cultured TIL used for patient treatment.


This resulted in the characterization of nine mutated epitopes from five of eight patients tested. Cells reactive with eight of the nine 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 patient peripheral blood prior to immune therapy. Moreover, we were able to rapidly isolate and clone from these cells TCRs specific for neoantigens that could be used to endow T cells with mutated antigens specificity. In addition, neo-antigens reactive T cells were detected in the patient peripheral blood for up to one year after treatment. We believe this potentially provides the basis for designing novel personalized immunotherapies for treating patients with advanced cancer.

Authors’ Affiliations

Bar-Ilan University, Ramat Gan, Israel
Surgery Branch/National Cancer Institute / National Institutes of Health, Bethesda, MD, USA
NIH/National Cancer Institute, Bethesda, MD, USA
NIH, Bethesda, MD, USA
NCI/NIH, Bethesda, MD, USA
NIH/NCI, Bethesda, MD, USA


© Cohen et al. 2015

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.