Therapeutic cancer peptide vaccination is an immunotherapy designed to elicit cytotoxic T-lymphocyte (CTL) responses in patients. A number of therapeutic vaccination trials have been performed, nevertheless there are only a few reports that have analyzed the T-cell receptors (TCRs) expressed on tumor antigen-specific CTLs. Here, we use next-generation sequencing (NGS) to analyze TCRs of vaccine-induced CTL clones and the TCR repertoire of bulk T cells in peripheral blood mononuclear cells (PBMCs) from two lung cancer patients over the course of long-term vaccine therapy. In both patients, vaccination with two epitope peptides derived from cancer/testis antigens (upregulated lung cancer 10 (URLC10) and cell division associated 1 (CDCA1)) induced specific CTLs expressing various TCRs. All URLC10-specific CTL clones tested showed Ca2+ influx, IFN-γ production, and cytotoxicity when co-cultured with URLC10-pulsed tumor cells. Moreover, in CTL clones that were not stained with the URLC10/MHC-multimer, the CD3 ζ chain was not phosphorylated. NGS of the TCR repertoire of bulk PBMCs demonstrated that the frequency of vaccine peptide-specific CTL clones was near the minimum detectable threshold level. These results demonstrate that vaccination induces antigen-specific CTLs expressing various TCRs at different time points in cancer patients, and that some CTL clones are maintained in PBMCs during long-term treatment, including some with TCRs that do not bind peptide/MHC-multimer.
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Cell division associated 1
CDCA1-derived HLA-A24 (A*24:02)-restricted peptide
CDCA1-64/ HLA-A*24:02 pentamer-PE
- CMV pp65 peptide:
CMV-derived HLA-A24 (A*24:02)-restricted peptide
- HIV epitope peptide:
HIV-derived HLA-A24 (A*24:02)-restricted peptide
Upregulated lung cancer 10
URLC10-derived HLA-A24 (A*24:02)-restricted peptide
URLC10-177/ HLA-A*24:02 tetramer-PE
Vascular endothelial growth factor receptor
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We are grateful to OncoTherapy Science, Inc. (Kanagawa, Japan) for their technical support.
This work was supported by the Ministry of Education, Science, and Culture, Japan (15K14410) to K. Takeda.
Conflict of interest
K. Kitaura and R. Suzuki are currently employed by Repertoire Genesis, Inc. The other authors declare that they have no conflicts of interest.
Ethical approval and ethical standards
This study was approved by the ethical committee of Fukushima Medical University (approval number: 810) and was registered with ClinicalTrials.gov (NCT00874588). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any animal studies performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
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Takeda, K., Kitaura, K., Suzuki, R. et al. Quantitative T-cell repertoire analysis of peripheral blood mononuclear cells from lung cancer patients following long-term cancer peptide vaccination. Cancer Immunol Immunother 67, 949–964 (2018). https://doi.org/10.1007/s00262-018-2152-x
- Therapeutic vaccine
- Cancer/testis antigen
- Next-generation sequencing