Roberti et al. found that vaccine site-infiltrating lymphocytes (VILs) isolated from a patient with TNBC who was treated with a personalized neoantigen peptide vaccine included both Trm-like CD4+ and CD8+ T cells with distinct antigen reactivities from PBMCs. One neoantigen (NCOR1L1475R) enabled CD8+ T cell expansion and formed stable HLA-I complexes, and TCR-T cells generated using reactive clonotypes responded to the neoantigen, but not the wild-type peptide. Post-vaccine tumor biopsy showed immune activation, but also loss of HLA-I expression and the NCOR1L1475R mutation, and NCOR1L1475R- reactive T cells were not found in the tumor or blood - only in VILs.
Contributed by Alex Najibi
ABSTRACT: T cells that recognize tumor-specific mutations are crucial for cancer immunosurveillance and in adoptive transfer of TILs or transgenic-TCR T cell products. However, their challenging identification and isolation limits their use in clinical practice. Therefore, novel approaches to isolate tumor-specific T cells are needed. Here, we report the isolation of neoantigen-specific CD8(+) T cells from a vaccination site of a metastatic breast cancer patient who received a personalized vaccine. Based on the somatic mutations, potential MHC binding epitopes were predicted, of which 17 were selected to generate a peptide vaccine. Cutaneous biopsies were processed after the fifth vaccination cycle to obtain infiltrating lymphocytes from the vaccination site (VILs). IFN_ ELISpot revealed reactivity to four peptides used in the vaccine. Reactive T cells from VILs were non-overlapping with those detected in the blood and the tumor-microenvironment. ScTCR Seq analysis revealed the presence of a clonotype in VILs that further expanded after a round of in vitro stimulation and validated to be specific against a private mutation, namely NCOR1(L1475R), presented in the context of HLA-B_*_07:02, with no reactivity to the wild-type peptide. Our study shows, for the first time, that tumor mutation - specific T cells are generated at high frequencies in the vaccination site and can be isolated with standard methods for TCR screening. The easy and safe accessibility of skin biopsies overcomes the major hurdles of current TCR screening approaches and present exciting opportunities for the development of innovative immunotherapeutic strategies.
Author Info: (1) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Inte
Author Info: (1) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. (2) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. Center for Quantitative Analysis of Molecular and Cellular Biosystems (Bioquant), Heidelberg University, Heidelberg, Germany. (3) Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. Center for Quantitative Analysis of Molecular and Cellular Biosystems (Bioquant), Heidelberg University, Heidelberg, Germany. (4) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. Antigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, Germany. (5) GMP and T Cell Therapy, German Cancer Research Center (DKFZ), Heidelberg, Germany. (6) Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. GMP and T Cell Therapy, German Cancer Research Center (DKFZ), Heidelberg, Germany. (7) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. Antigen Presentation and T/NK Cell Activation Group, German Cancer Research Center (DKFZ), Heidelberg, Germany. (8) Systems Immunology and Single Cell Biology Group, German Cancer Research Center (DKFZ), Heidelberg, Germany. Faculty of Biosciences, Heidelberg University, Heidelberg, Germany. (9) Systems Immunology and Single Cell Biology Group, German Cancer Research Center (DKFZ), Heidelberg, Germany. German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), Core Center Heidelberg, Heidelberg, Germany. (10) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. Center for Quantitative Analysis of Molecular and Cellular Biosystems (Bioquant), Heidelberg University, Heidelberg, Germany. (11) Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany. (12) Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. Division of Gynecological Oncology, National Center for Tumor Diseases (NCT), A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg University Hospital, Heidelberg, Germany. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany. (13) National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany. (14) Division of Gynecological Oncology, National Center for Tumor Diseases (NCT), A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg University Hospital, Heidelberg, Germany. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany. (15) National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. Division of Gynecological Oncology, National Center for Tumor Diseases (NCT), A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg University Hospital, Heidelberg, Germany. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany. (16) Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. Division of Gynecological Oncology, National Center for Tumor Diseases (NCT), A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg University Hospital, Heidelberg, Germany. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany. (17) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany. (18) Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany. Medical Faculty Heidelberg, Department of Medical Oncology and Internal Medicine VI, Heidelberg University Hospital, Heidelberg University, Heidelberg, Germany. National Center for Tumor Diseases (NCT) Heidelberg, A partnership between DKFZ and Heidelberg University Medical Center, Heidelberg, Germany.
