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Isolation of a tumor neoantigen specific CD8+ TCR from a skin biopsy of a vaccination site
Maria Paula Roberti1,2,3, Pornpimol Charoentong1,2,3,4, Yanhong Lyu2,4
1Clinical Cooperation Unit Applied Tumor Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany.
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). IFNγ 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 NCOR1L1475R, 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.
Insights
Researchers isolated tumor mutation-specific T cells from a personalized cancer vaccine site. This breakthrough in T cell isolation from skin biopsies offers new avenues for cancer immunotherapy development.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- T cells recognizing tumor-specific mutations are vital for cancer immunity and adoptive cell therapies.
- Identifying and isolating these tumor-specific T cells is challenging, hindering clinical applications.
- Novel methods are required to effectively isolate tumor-specific T cells for therapeutic use.
Purpose of the Study:
- To report the successful isolation of neoantigen-specific CD8+ T cells from a vaccination site in a metastatic breast cancer patient.
- To demonstrate a new approach for isolating tumor mutation-specific T cells using accessible skin biopsies.
- To explore the potential of vaccination-site T cells for developing innovative immunotherapies.
Main Methods:
- Personalized peptide vaccine generation based on predicted MHC-binding neoantigens from somatic mutations.
- Isolation of tumor-infiltrating lymphocytes from the vaccination site (VILs) via cutaneous biopsies.
- IFNγ ELISpot assay for T cell reactivity, ScTCR Seq for clonotype analysis, and in vitro stimulation for T cell expansion and validation.
Main Results:
- Four vaccine peptides elicited T cell reactivity in VILs, distinct from blood and tumor-infiltrating lymphocytes.
- A specific T cell receptor (TCR) clonotype targeting a private mutation (NCOR1L1475R) presented by HLA-B*07:02 was identified in VILs.
- This identified TCR demonstrated specificity for the mutated peptide over the wild-type, with high frequency and expansion in VILs.
Conclusions:
- Tumor mutation-specific T cells are generated at high frequencies within the vaccination site following personalized vaccination.
- Standard methods can successfully isolate these T cells from skin biopsies, overcoming current TCR screening limitations.
- Accessible skin biopsies offer a promising strategy for isolating tumor-specific T cells, advancing cancer immunotherapy development.
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