Personalized ex vivo multiple peptide enrichment and detection of T cells reactive to multiple tumor-associated antigens in prostate cancer patients
Personalized peptide vaccination is a promising immunotherapeutic approach in prostate cancer (PCa). We therefore examined whether an approach, utilizing personalized multiple peptide-mediated ex vivo enrichment with effector T cells reactive to multiple tumor-associated antigens (TAAs), could be employed as a basis for the development of T cell immunotherapy of PCa. In this study, we used the non-adherent fraction (lymphocytes) of cryopreserved peripheral blood mononuclear cells from a leukapheretic product of biochemically recurrent (BR, n = 14) and metastatic hormone-refractory (HR, n = 12) PCa patients. The lymphocytes were primed with a pool of mixed overlapping peptides derived from 6 PCa TAAs–PSA, PAP, NY-ESO-1, MAGE-A1, MAGE-A3 and MAGE-A4. After 2 weeks of culture, the cells were stimulated with the peptides and T cell reactivity determined by externalization of CD107a. No TAAs-reactive effector T cells were detected in the patient’s lymphocytes after their reconstitution. However, following their priming with the TAAs-derived peptides and 2-week culturing, the lymphocytes became enriched with polyclonal TAAs-reactive effector CD8+ T cells in 8 out of 14 BR and 5 out of 12 HR patients. No such reactive CD8+ T cells were detected in cultured lymphocytes without the peptide priming. Stimulation of the responding cultures with peptides derived from individual TAAs revealed a unique repertoire of the reactive CD8+ T cells. Our strategy revealed that the personalized multiple peptide-mediated ex vivo enrichment with multiple TAAs-reactive T cells in the PCa patient’s lymphocytes is a viable approach for development of T cell immunotherapy of PCa.
KeywordsProstate cancer Tumor-associated antigens Personalized T cell immunotherapy CD107a externalization
Adoptive cell transfer
Prostatic acid phosphatase
Prostate-specific membrane antigen
Personalized peptide vaccination
Peripheral blood mononuclear cells
New York esophageal squamous cell carcinoma-1
Biochemically recurrent prostate cancer
Hormone-refractory prostate cancer
We would like to thank the clinical research staff, routine laboratory staff and our technicians for their assistance. We thank Dr. Alasdair M. Gilfillan for a critical review of the manuscript.
Research in the authors’ laboratories was supported by funding of the Charles University—Project GA UK No. 188215 and PRIMUS/MED/12, the Ministry of Health, Czech Republic—Project AZV 16-28135A and the conceptual development fund of research organization University Hospital Motol, Prague, Czech Republic 00064203.
Compliance with ethical standards
Conflict of interest
JB a MP are part-time employees of SOTIO, a.s., a biotech company developing cell-based immunotherapy. JB is a minority shareholder of this company. PT, ZS, KV, Dmitry Stakheev and Daniel Smrz declare no financial or commercial conflict of interest.
This study was approved by the ethics committee for multicentric studies and evaluation of the Faculty Hospital Motol, Prague, Czech Republic. 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.
Informed consent was obtained from all individual participants included in the study.
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