Skip to main content
SpringerLink
Log in

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

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Collin M, Bigley V (2018) Human dendritic cell subsets: an update. Immunology 154(1):3–20. https://doi.org/10.1111/imm.12888

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  2. Cancel J-C, Crozat K, Dalod M, Mattiuz R (2019) Are conventional type 1 dendritic cells critical for protective antitumor Immunity and How? Front Immunol 10(19):9–9. https://doi.org/10.1038/s41467-019-13368-y (ISSN 1664-3224)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  3. Eisenbarth SC (2019) Dendritic cell subsets in T cell programming: location dictates function. Nat Rev Immunol 19(2):89–103. https://doi.org/10.1038/s41577-018-0088-1.ISSN1474-1733

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Cintolo JA, Datta J, Mathew SJ, Czerniecki BJ (2012) Dendritic cell-based vaccines: barriers and opportunities. Future Oncol 8(10):1273–1299. https://doi.org/10.2217/fon.12.125 (ISSN 1479-6694)

    Article  PubMed  CAS  Google Scholar 

  5. Yu J, Sun H, Cao W, Song Y, Jiang Z (2022) Research progress on dendritic cell vaccines in cancer immunotherapy. Exp Hematol Oncol 11(1):3. https://doi.org/10.1186/s40164-022-00257-2

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. PantelSpeicher KMR (2016) The biology of circulating tumor cells. Oncogene 35(10):1216–1224. https://doi.org/10.1038/onc.2015.192.ISSN0950-9232

    Article  Google Scholar 

  7. Kolostova K, Zhang Y, Hoffman RM, Bobek V (2014) In vitro culture and characterization of human lung cancer circulating tumor cells isolated by size exclusion from an orthotopic nude-mouse model expressing fluorescent protein. J Fluorescence 24(5):1531–1536. https://doi.org/10.1007/s10895-014-1439-3 (ISSN 1053-0509)

    Article  CAS  Google Scholar 

  8. Kolostova K, Matkowski R, Jędryka M et al (2015) The added value of circulating tumor cells examination in ovarian cancer staging. Am J Cancer Res 5(11):3363–3375

    PubMed  PubMed Central  CAS  Google Scholar 

  9. Nath SC, Harper L, Rancourt DE (2020) Cell-based therapy manufacturing in stirred suspension bioreactor: thoughts for cGMP compliance. Front Bioeng Biotechnol 8(8):1–16. https://doi.org/10.3389/fbioe.2020.599674 (ISSN 2296-4185)

    Article  Google Scholar 

  10. Hopewell EL, Cox C (2020) Manufacturing dendritic cells for immunotherapy: monocyte enrichment. Mol Ther Methods Clin Dev 16(1):155–160. https://doi.org/10.1016/j.omtm.2019.12.017 (ISSN 23290501)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Perez CR, De Palma M (2019) Engineering dendritic cell vaccines to improve cancer immunotherapy. Nat Commun 10(1):1–10. https://doi.org/10.1038/s41467-019-13368-y (ISSN 2041-1723)

    Article  CAS  Google Scholar 

  12. Silvestris N, Ciliberto G, De Paoli P, Apolone G, Lavitrano ML, Pierotti MA, Stanta G (2017) Liquid dynamic medicine and N-of-1 clinical trials a change of perspective in oncology research. J Exp Clin Cancer Res 36(1):5. https://doi.org/10.1186/s13046-017-0598-x (ISSN 1756-9966)

    Article  Google Scholar 

  13. Pantel K, Alix-Panabières C (2019) Liquid biopsy and minimal residual disease—latest advances and implications for cure. Nat Rev Clin Oncol 16(7):409–424. https://doi.org/10.1038/s41571-019-0187-3 (ISSN 1759-4774)

    Article  PubMed  CAS  Google Scholar 

  14. Heiser A, Coleman D, Dannull J et al (2002) Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors. J Clin Investig 109(3):409–417. https://doi.org/10.1172/JCI14364 (ISSN 0021-9738)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Saxena M, Balan S, Roudko V, Bhardwaj N (2018) Towards superior dendritic-cell vaccines for cancer therapy. Nat Biomed Eng 2(6):341–346. https://doi.org/10.1038/s41551-018-0250-x (ISSN 2157-846X)

    Article  PubMed  PubMed Central  Google Scholar 

  16. Koido S, Kashiwaba M, Chen D, Gendler S, Kufe D, Gong J (2000) Induction of antitumor immunity by vaccination of dendritic cells transfected with MUC1 RNA. J Immunol 165(10):5713–5719. https://doi.org/10.4049/jimmunol.165.10.5713 (ISSN 0022-1767)

    Article  PubMed  CAS  Google Scholar 

  17. Wculek SK, Cueto FJ, Mujal AM, Melero I, Krummel MF, Sancho D (2020) Dendritic cells in cancer immunology and immunotherapy. Nat Rev Immunol 20(1):7–24. https://doi.org/10.1038/s41577-019-0210-z (ISSN 1474-1733)

    Article  PubMed  CAS  Google Scholar 

  18. Constantino J, Gomes C, Falcão A, Cruz MT, Neves BM (2016) Antitumor dendritic cell–based vaccines: lessons from 20 years of clinical trials and future perspectives. Transl Res 168:74–95. https://doi.org/10.1016/j.trsl.2015.07.008 (ISSN 19315244)

    Article  PubMed  CAS  Google Scholar 

  19. Amberger DC, Schmetzer HM (2020) Dendritic cells of leukemic origin: specialized antigen-presenting cells as potential treatment tools for patients with Myeloid Leukemia. Transfus Med Hemother 47(6):432–443. https://doi.org/10.1159/000512452 (ISSN 1660-3796)

    Article  PubMed  PubMed Central  Google Scholar 

  20. Sauter B, Albert ML, Francisco L, Larsson M, Somersan S, Bhardwaj N (2000) Consequences of Cell Death. J Exp Med 191(3):423–434. https://doi.org/10.1084/jem.191.3.423 (ISSN 0022-1007)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Podrazil M, Horvath R, Becht E et al (2015) Phase I/II clinical trial of dendritic-cell based immunotherapy (DCVAC/PCa) combined with chemotherapy in patients with metastatic, castration-resistant prostate cancer. Oncotarget 6(20):18192–18205. https://doi.org/10.18632/oncotarget.4145 (ISSN 1949-2553)

    Article  PubMed  PubMed Central  Google Scholar 

  22. Oh SA, Wu DC, Cheung J (2020) PD-L1 expression by dendritic cells is a key regulator of T-cell immunity in cancer. Nat Cancer 1(1):681–691. https://doi.org/10.1038/s43018-020-0075-x

    Article  PubMed  CAS  Google Scholar 

  23. Morita M, Gravel S-P, Hulea L, Larsson O, Pollak M, St-Pierre J, Topisirovic I (2015) MTOR coordinates protein synthesis, mitochondrial activity and proliferation. Cell Cycle 14(4):473–480. https://doi.org/10.4161/15384101.2014.991572 (ISSN 1538-4101)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Szatmari I, Töröcsik D, Agostini M, Nagy T, Gurnell M, Barta E, Chatterjee K, Nagy L (2007) PPARγ regulates the function of human dendritic cells primarily by altering lipid metabolism. Blood 110(9):3271–3280. https://doi.org/10.1182/blood-2007-06-096222 (ISSN 0006-4971)

    Article  PubMed  CAS  Google Scholar 

  25. Pearce EJ, Everts B (2015) Dendritic cell metabolism. Nat Rev Immunol 15(1):18–29. https://doi.org/10.1038/nri3771 (ISSN 1474-1733)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  26. Krawczyk CM, Holowka T, Sun J et al (2010) Toll-like receptor–induced changes in glycolytic metabolism regulate dendritic cell activation. Blood 115(23):4742–4749. https://doi.org/10.1182/blood-2009-10-249540 (ISSN 0006-4971)

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

Acknowledgements

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

Author information

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
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eliska Pospisilova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rafal Matkowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jolanta Szelachowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vladimir Bobek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vladimir Bobek.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (XLSX 4369 kb)

Supplementary file1 (DOCX 18 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

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

Keywords

Search

Navigation