Whiteside Theresa L, Gambotto Andrea, Albers Andreas, Stanson Joanna, Cohen Edward P
Department of Pathology, University of Pittsburgh School of Medicine and University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Proc Natl Acad Sci U S A. 2002 Jul 9;99(14):9415-20. doi: 10.1073/pnas.142302399. Epub 2002 Jun 21.
This article describes a DNA-based vaccination strategy evaluated ex vivo with human cells. The vaccine was prepared by transferring tumor-derived genomic DNA to PCI-13 cells, a highly immunogenic tumor cell line ("recipient cell"), which had been genetically modified to secrete IL-2 (PCI-13/IL-2). PCI-13 cells expressed class I MHC determinants (HLA-A2) shared with the tumor from which the DNA was obtained as well as allogeneic determinants. DNA from a gp100(+) melanoma cell line was transduced into gp100(-) PCI-13/IL-2 cells (PCI-13/IL-2/DNA). A T cell line specific for the gp100 epitope responded to PCI-13/IL-2/DNA cells by IFN-gamma-secretion measured in enzyme-linked immunospot assays. The T cell line also recognized the gp100 epitope presented by dendritic cells that ingested PCI-13/IL-2/DNA cells, which had been induced by UVB irradiation to undergo apoptosis. After up-take and processing of apoptotic PCI-13/IL-2/DNA cells, the dendritic cells primed normal peripheral blood lymphocytes to generate effector T cells specific for the tumor donating the DNA. The results indicate that tumor epitopes encoded in such DNA are expressed in recipient cells and can induce tumor-specific T cells. The findings support translation of this vaccination strategy to a phase I trial in patients with cancer.
本文描述了一种基于DNA的疫苗接种策略，该策略已在体外用人细胞进行了评估。疫苗是通过将肿瘤来源的基因组DNA转移到PCI-13细胞（一种高度免疫原性的肿瘤细胞系，即“受体细胞”）中制备的，该细胞系已被基因改造以分泌IL-2（PCI-13/IL-2）。PCI-13细胞表达与获取DNA的肿瘤共有的I类MHC决定簇（HLA-A2）以及同种异体决定簇。来自gp100(+)黑色素瘤细胞系的DNA被转导到gp100(-)的PCI-13/IL-2细胞中（PCI-13/IL-2/DNA）。在酶联免疫斑点试验中，对gp100表位特异的T细胞系通过分泌IFN-γ对PCI-13/IL-2/DNA细胞作出反应。该T细胞系还识别摄取了经UVB照射诱导凋亡的PCI-13/IL-2/DNA细胞的树突状细胞所呈递的gp100表位。在摄取并处理凋亡的PCI-13/IL-2/DNA细胞后，树突状细胞使正常外周血淋巴细胞致敏，以产生对捐赠DNA的肿瘤特异的效应T细胞。结果表明，此类DNA中编码的肿瘤表位在受体细胞中表达，并可诱导肿瘤特异性T细胞。这些发现支持将这种疫苗接种策略转化为针对癌症患者的I期试验。
