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Immune activation of the monocyte-derived dendritic cells using patients own circulating tumor cells

Cancer Immunology, Immunotherapy volume 71, pages 2901–2911 (2022)Cite this article

Abstract

Background

Dendritic cell (DC) therapy counts to the promising strategies how to weaken and eradicate cancer disease. We aimed to develop a good manufacturing practice (GMP) protocol for monocyte-derived DC (Mo-DC) maturation using circulating tumor cells lysates with subsequent experimental T-cell priming in vitro.

Methods

DC differentiation was induced from a population of immunomagnetically enriched CD14 + monocytes out of the leukapheresis samples (n = 6). The separation was provided automatically, in a closed bag system, using CliniMACS Prodigy® separation protocols (Miltenyi Biotec). For differentiation and maturation of CD14 + cells, DendriMACs® growing medium with supplements (GM-CSF, IL-4, IL-6, IL-1B, TNFa, PGE) was used. Immature Mo-DCs were loaded with autologous circulating tumor cell (CTCs) lysates. Autologous CTCs were sorted out by size-based filtration (MetaCell®) of the leukapheresis CD14-negative fraction. A mixture of mature Mo-DCs and autologous non-target blood cells (NTBCs) was co-cultured and the activation effect of mature Mo-DCs on T-cell activation was monitored by means of multimarker gene expression profiling.

Results

New protocols for mMo-DC production using automatization and CTC lysates were introduced including a feasible in vitro assay for mMo-DC efficacy evaluation. Gene expression analysis revealed elevation for following genes in NTBC (T cells) subset primed by mMo-DCs: CD8A, CD4, MKI67, MIF, TNFA, CD86, and CD80 (p ≤ 0.01).

Conclusion

Summarizing the presented data, we might conclude mMo-DCs were generated using CliniMACS Prodigy® machine and CTC lysates in a homogenous manner showing a potential to generate NTBC activation in co-cultures. Identification of the activation signals in T-cell population by simple multimarker-qPCRs could fasten the process of effective mMo-DC production.

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Acknowledgements

This study was supported by Krajska zdravotni, a.s, Grant nr.: IGA-KZ-2017-1-16. The figures were created with BioRender.com.

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Authors and Affiliations

  1. Laboratory of Personalized Medicine, Oncology Clinic, University Hospital Kralovske Vinohrady, Srobarova 50, 10034, Prague, Czech Republic

    Katarina Kolostova, Eliska Pospisilova & Vladimir Bobek

  2. Department of Oncology, Wrocław Medical University, Wrocław, Poland

    Rafal Matkowski & Jolanta Szelachowska

  3. Breast Cancer Unit, Lower Silesian Oncology, Pulmonology and Hematology Center, Plac Hirszfelda 12, 54-413, Wrocław, Poland

    Rafal Matkowski & Jolanta Szelachowska

  4. 3rd Department of Surgery University Hospital Motol and 1st Faculty of Medicine, Charles University, V Uvalu 84, 15006, Prague, Czech Republic

    Vladimir Bobek

  5. Department of Thoracic Surgery, Masaryk’s Hospital, Krajska Zdravotni a.s., Socialni pece 3316/12A, 40113, Usti nad Labem, Czech Republic

    Vladimir Bobek

  6. Department of Thoracic Surgery, Lower Silesian Oncology, Pulmology and Hematology Center and Medical University Wroclaw, Grabiszynska 105, 53-413, Wrocław, Poland

    Vladimir Bobek

Authors
  1. Katarina Kolostova
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Correspondence to Vladimir Bobek.

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Kolostova, K., Pospisilova, E., Matkowski, R. et al. Immune activation of the monocyte-derived dendritic cells using patients own circulating tumor cells. Cancer Immunol Immunother 71, 2901–2911 (2022). https://doi.org/10.1007/s00262-022-03189-2

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  • DOI: https://doi.org/10.1007/s00262-022-03189-2

Keywords

  • Dendritic cells
  • Immunotherapy
  • T cells
  • Personalized medicine
  • MetaCell
  • Circulating tumor cells