Korean J Hematol.
2001 Aug;36(3):223-231.
Induction of Cytotoxic T Lymphocytes by Dendritic Cells Pulsed with Murine Leukemic Cell RNA
- Affiliations
-
- 1Division of Hematology Oncology1, Department of Medicine, Samsung Medical Center, Seoul, Korea.
- 2Department of Internal Medicine, College of Medicine, Chung-Ang University, Seoul, Korea.
- 3Cancer Research Center, Seoul National University College of Medicine, Seoul, Korea.
- 4Department of Internal Medicine, College of Medicine, Chungbuk National University, Cheongju, Chungbuk, Korea.
- 5Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea.
- 6Department of Internal Medicine, Hanyang University School of Medicine, Seoul, Korea.
Abstract
- BACKGROUND
This study was done to assess the feasibility of dendritic cell generation from murine bone marrow and the efficacy of dendritic cells pulsed with total RNA to induce specific cytotoxic T lymphocyte response against leukemic cells.
METHODS
Nucleated cells of inbred BALB/c mice were obtained and cultured with granulocyte/macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) to induce dendritic cells. Total RNA of WEHI-3BD+, a myelomonocytic leukemia cell line from BALB/c, was transfected into the dendritic cells using liposome. RNA pulsed dendritic cells were irradiated and administered to the BALB/c mice intraperitoneally and splenic T lymphocytes were harvested. After restimulation with leukemic cells, T cell proliferation and specific cytotoxicity was assessed.
RESULTS
Cells cultured with GM-CSF and lipopolysaccaride were found to have prominent dendritic processes. The percentage of cells showing high expression of both MHC class II and CD80, CD86, or CD11c was 69.6 %, 63.7%, and 41.8%, respectively. T cells stimulated by WEHI-3BD+ total RNA pulsed dendritic cells using DOTAP showed enhanced proliferation than those stimulated by total RNA or media only (P=0.05). When T cells were cocultured with WEHI-3BD+ as target cells, T cells stimulated by WEHI-3BD+ total RNA pulsed dendritic cells using DOTAP showed much increased cytotoxicity than controls.
CONCLUSION
Dendritic cells pulsed with total leukemic RNA could stimulate T cells to induce specific cytotoxic effect.
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