Enhanced Dendritic Cell Antigen Presentation in RNA-Based Immunotherapy
| Autor: | Scott K. Pruitt, Karen M. Padilla, Douglas S. Tyler, Sirisha Emani, Mark W. Onaitis, Matthew F. Kalady |
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| Rok vydání: | 2002 |
| Předmět: |
T-Lymphocytes
medicine.medical_treatment Antigen presentation Biology Transfection Cell Line Interferon-gamma Antigen medicine Humans RNA Messenger Cellular Senescence Antigen Presentation Vaccines Follicular dendritic cells Antigen processing Dendritic Cells Immunotherapy Dendritic cell Molecular biology Cell biology Electroporation Surgery Cell aging |
| Zdroj: | Journal of Surgical Research. 105:17-24 |
| ISSN: | 0022-4804 |
| Popis: | Dendritic cells pulsed with mRNA provide a unique approach to tumor immunotherapy. We hypothesized that increased mRNA transfection efficiency and dendritic cell maturation would improve antigen processing and presentation as well as T-cell costimulation, resulting in enhanced induction of antimelanoma immune responses.Immature monocyte-derived dendritic cells were transfected with mRNA by passive pulsing, lipofection, or electroporation. Dendritic cells were either left untreated or matured using the double-stranded RNA poly(I:C). T-Cell cultures were generated by stimulation of naïve T-cells with each set of dendritic cells. Specific antigen presentation and specific effector T-cell generation were analyzed by an IFN-gamma release Elispot assay.Greatest intracellular green fluorescent protein was observed by flow cytometry following dendritic cell electroporation with green fluorescent protein mRNA. DC presentation of Mart-1/Melan A peptide, as measured by Elispot assay using a specific T-cell clone, was greatest following transfection with Mart-1/Melan A mRNA by electroporation. Maturation of dendritic cells further improved antigen presentation regardless of transfection technique. Specific Mart-1/Melan A effector T cells were produced after culture of naïve T cells with dendritic cells that were electroporated with Mart-1/Melan A mRNA and then matured, but not for dendritic cells that remained immature.Efficient mRNA transfection by electroporation as well as dendritic cell maturation results in increased levels of Mart-1/Melan A antigen presentation and enhanced production of antigen-specific effector T cells. This combination of strategies may be used to enhance immune responses to RNA-based dendritic cell vaccines. |
| Databáze: | OpenAIRE |
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