Induction of antigen-specific CD8 cytotoxic T cells by dendritic cells co-electroporated with a dsRNA analogue and tumor antigen mRNA

• Published in: Gene therapy Vol. 13; no. 13; pp. 1027 - 1036
• Main Authors: MICHIELS, A, BRECKPOT, K, CORTHALS, I, TUYAERTS, S, BONEHILL, A, HEIRMAN, C, THIELEMANS, K, AERTS, J. L
• Format: Journal Article
• Language: English
• Published: Basingstoke Nature Publishing Group 01.07.2006
• Subjects: Adaptive immunity; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Antigen Presentation; Antigens; Antigens, Neoplasm - genetics; Antigens, Neoplasm - immunology; Applied cell therapy and gene therapy; Biological and medical sciences; Biotechnology; CD8 antigen; CD8-Positive T-Lymphocytes - immunology; Cell activation; Cytokines; Cytotoxicity; Cytotoxicity Tests, Immunologic; Dendritic cells; Dendritic Cells - immunology; Double-stranded RNA; Electroporation; Electroporation - methods; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gene therapy; Genetic aspects; Genetic Engineering; Genetic Therapy - methods; Health aspects; Health. Pharmaceutical industry; Humans; Immunology; Immunotherapy; Immunotherapy - methods; Industrial applications and implications. Economical aspects; Inflammation; Interferon; Interferon-gamma - immunology; Lymphocyte Activation; Lymphocytes T; MART-1 Antigen; Medical sciences; Melanoma, Experimental - immunology; Melanoma, Experimental - therapy; Methods; Molecular biology; Monocytes; mRNA; Mutation; Neoplasm Proteins - genetics; Physiological aspects; RNA - administration & dosage; RNA, Double-Stranded - administration & dosage; Skin Neoplasms - immunology; Skin Neoplasms - therapy; T cells; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Tumor Cells, Cultured; Tumors; Belgium; dsRNA; Dendritic cell; Induction; CTL induction; Cytotoxicity; Electroporation; RNA electroporation; Antigen; maturation; Messenger RNA; Immunotherapy; T-Lymphocyte; Tumor; Gene therapy
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/sj.gt.3302750

The maturation state of dendritic cells (DCs) is an important determinant for the initiation and regulation of adaptive immune responses. In this study, we wanted to assess whether functional activation of human monocyte-derived DCs can be achieved by electroporation of an activation signal in the form of double-stranded (ds) RNA and whether simultaneous electroporation of the dsRNA with tumor antigen encoding mRNA can lead to the induction of a cytotoxic T-lymphocyte (CTL) response. Electroporation of immature DCs with poly(I:C(12)U), a dsRNA analogue, resulted in phenotypic as well as functional changes, indicative of DC maturation. Co-electroporation of DCs with both poly(I:C(12)U) and Melan-A/MART-1 encoding mRNA induced strong anti-Melan-A/MART-1 CD8(+) T-cell responses in vitro. Higher numbers of Melan-A/MART-1-specific CTLs were consistently obtained with poly(I:C(12)U)-activated DCs compared to DCs matured in the presence of an inflammatory cytokine cocktail. These results indicate that DC co-electroporation with both dsRNA and tumor antigen encoding mRNA induces fully activated and antigen-loaded DCs that promote antigen-specific CTL responses and may provide the basis for future immunotherapeutic strategies.