The natural adjuvant properties of bacterial ghosts (BGs) lie within the presence of intact pathogen-associated molecular patterns on their surface. BGs can improve the direct delivery, natural processing and presentation of target antigens within dendritic cells (DCs). Moreover, sensitization of human DCs by cancer cell lysate (oncolysate)-loaded BGs in the presence of IFN-α and GM-CSF enhanced DC maturation as indicated by an increased expression of maturation markers and co-stimulatory molecules, higher production of IL-12p70 and stimulation of significantly increased proliferation of both autologous CD4+ and CD8+ T cells compared to DCs matured in the presence of purified lipopolysaccharide. The induced T cells efficiently recognized oncolysate-derived tumor-associated antigens expressed by cancer cells used for the production of oncolysate. Our optimized one-step simultaneous antigen delivery and DC maturation-inducing method emerges as a promising tool for the development and implementation of next-generation cellular cancer immunotherapies.

• Keywords
• Bacterial ghosts, Cancer immunotherapy, Dendritic cells, Natural adjuvant, T cell stimulation, Vaccine,
• MeSH
• Cell Differentiation immunology   MeSH
• CD4-Positive T-Lymphocytes immunology   MeSH
• CD8-Positive T-Lymphocytes immunology   MeSH
• Dendritic Cells immunology   microbiology   transplantation   MeSH
• Escherichia coli immunology   MeSH
• Phenotype MeSH
• Glioblastoma immunology   therapy   MeSH
• Immunotherapy, Adoptive methods   MeSH
• Interleukin-12 biosynthesis   immunology   MeSH
• Humans MeSH
• Lipopolysaccharides pharmacology   MeSH
• Cell Line, Tumor MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Interleukin-12 MeSH
• Lipopolysaccharides MeSH

Dendritic cells (DCs) belong to the immune system and are particularly studied for their potential to direct either an activated or tolerogenic immune response. The roles of microRNAs (miRNAs) in posttranscriptional gene expression regulation are being increasingly investigated. This study's aim is to evaluate the miRNAs' expression changes in prepared human immature (iDCs), activated (aDCs), and tolerogenic dendritic cells (tDCs). The dendritic cells were prepared using GM-CSF and IL-4 (iDC) and subsequently maturated by adding LPS and IFN-γ (aDC) or IL-10 and TGF-β (tDC). Surface markers, cytokine profiles, and miRNA profiles were evaluated in iDC, tDC, and aDC at 6 h and 24 h of maturation. We identified 4 miRNAs (miR-7, miR-9, miR-155 and miR-182), which were consistently overexpressed in aDC after 6 h and 24 h of maturation and 3 miRNAs (miR-17, miR-133b, and miR-203) and miR-23b cluster solely expressed in tDC. We found 5 miRNAs (miR-10a, miR-203, miR-210, miR-30a, and miR-449b) upregulated and 3 miRNAs downregulated (miR-134, miR-145, and miR-149) in both tDC and aDC. These results indicate that miRNAs are specifically modulated in human DC types. This work may contribute to identifying specific modulating miRNAs for aDC and tDC, which could in the future serve as therapeutic targets in the treatment of cancer and autoimmune diseases.

• MeSH
• Dendritic Cells drug effects   metabolism   MeSH
• Granulocyte-Macrophage Colony-Stimulating Factor pharmacology   MeSH
• Interleukin-10 pharmacology   MeSH
• Interleukin-4 pharmacology   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Lipopolysaccharides pharmacology   MeSH
• MicroRNAs genetics   MeSH
• Flow Cytometry MeSH
• Transforming Growth Factor beta pharmacology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Granulocyte-Macrophage Colony-Stimulating Factor MeSH
• Interleukin-10 MeSH
• Interleukin-4 MeSH
• Lipopolysaccharides MeSH
• MicroRNAs MeSH
• MIRN10 microRNA, human MeSH Browser
• MIRN133 microRNA, human MeSH Browser
• MIRN17 microRNA, human MeSH Browser
• MIRN203 microRNA, human MeSH Browser
• MIRN210 microRNA, human MeSH Browser
• MIRN30b microRNA, human MeSH Browser
• MIRN449 microRNA, human MeSH Browser
• Transforming Growth Factor beta MeSH