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Blood, 1 March 2004, Vol. 103, No. 5, pp. 1763-1769.
Prepublished online as a Blood First Edition Paper on November 13, 2003; DOI 10.1182/blood-2003-06-2097.
Submitted June 26, 2003
Accepted October 29, 2003
Induction of chronic lymphocytic leukemia (CLL)-specific CD4- and CD8-mediated T-cell responses using RNA-transfected dendritic cells
Martin R Mueller, Garyfalia Tsakou, Frank Gruenebach, Susanne M Schmidt, and Peter Brossart*
Department of Hematology, Oncology and Immunology, University of Tuebingen, Tuebingen, Germany
* Corresponding author; email: peter.brossart{at}med.uni-tuebingen.de.
Recently, it was demonstrated that transfection of DC with tumor-derived RNA can elicit effective T cell responses. This technique does not require the definition of the tumor antigen or HLA haplotype of the patients. We applied this approach to induce HLA class I- and class II- restricted T cell responses directed against malignant cells from patients with chronic lymphocytic leukemia (B-CLL). Here, we show that DC generated from monocytes of patients with B-CLL induce leukemia-specific cytotoxic and proliferative T cell responses upon transfection with total RNA isolated from autologous leukemic B lymphocytes. Standard 51Cr-release assays showed specific MHC class I restricted cytotoxic activity against the autologous leukemic B cells and DC transfected with CLL-RNA, whereas non-malignant B cells were spared. The specificity of the CTL response was confirmed using cold target inhibiton assays and by blocking HLA class I molecules. Furthermore, we established a protocol for the amplification of whole B-CLL-mRNA. The use of DC transfected with in vitro amplified B-CLL-mRNA elicited specific T cell responses similar to the results obtained with native mRNA. These data suggest that vaccinations using DC transfected with RNA might be a potent new strategy in the treatment of CLL.
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