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

• Published in: Cancer Immunology, Immunotherapy Immunotherapy, 2022-12, Vol.71 (12), p.2901-2911
• Main Authors: Kolostova, Katarina, Pospisilova, Eliska, Matkowski, Rafal, Szelachowska, Jolanta, Bobek, Vladimir
• Format: Article
• Language: English
• Subjects: Automation; Blood cells; Cancer Research; CD14 antigen; CD4 antigen; CD80 antigen; CD86 antigen; Cell activation; Cell differentiation; Dendritic cells; Dendritic Cells - metabolism; Gene expression; Good Manufacturing Practice; Granulocyte-macrophage colony-stimulating factor; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Humans; IL-1β; Immunology; Interleukin 1; Interleukin 4; Interleukin 6; Interleukin-4 - pharmacology; Interleukin-6 - pharmacology; Kinases; Leukapheresis; Lymphocytes T; Lysates; Maturation; Medicine; Medicine & Public Health; Monocytes; Monocytes - metabolism; Neoplastic Cells, Circulating - metabolism; Oncology; Original; Original Article; Prostaglandins E - pharmacology; Tumor cells
• ISSN: 0340-7004; 1432-0851
• DOI: 10.1007/s00262-022-03189-2

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.