Thirukkumaran Chandini M, Luider Joanne M, Stewart Douglas A, Cheng Tina, Lupichuk Sasha M, Nodwell Michael J, Russell James A, Auer Iwona A, Morris Donald G
Calgary Laboratory Services, Calgary, Alberta, Canada.
Blood. 2003 Jul 1;102(1):377-87. doi: 10.1182/blood-2002-08-2508. Epub 2003 Mar 13.
Hematologic stem cell rescue after high-dose cytotoxic therapy is extensively used for the treatment of many hematopoietic and solid cancers. Gene marking studies suggest that occult tumor cells within the autograft may contribute to clinical relapse. To date purging of autografts contaminated with cancer cells has been unsuccessful. The selective oncolytic property of reovirus against myriad malignant histologies in in vitro, in vivo, and ex vivo systems has been previously demonstrated. In the present study we have shown that reovirus can successfully purge cancer cells within autografts. Human monocytic and myeloma cell lines as well as enriched ex vivo lymphoma, myeloma, and Waldenström macroglobulinemia patient tumor specimens were used in an experimental purging model. Viability of the cell lines or purified ex vivo tumor cells of diffuse large B-cell lymphoma, chronic lymphocytic leukemia, Waldenström macroglobulinemia, and small lymphocytic lymphoma was significantly reduced after reovirus treatment. Further, [35S]-methionine labeling and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of cellular proteins demonstrated reovirus protein synthesis and disruption of host cell protein synthesis as early as 24 hours. Admixtures of apheresis product with the abovementioned tumor cells and cell lines treated with reovirus showed complete purging of disease. In contrast, reovirus purging of enriched ex vivo multiple myeloma, Burkitt lymphoma, and follicular lymphoma was incomplete. The oncolytic action of reovirus did not affect CD34+ stem cells or their long-term colony-forming assays even after granulocyte colony-stimulating factor (G-CSF) stimulation. Our results indicate the ex vivo use of an unattenuated oncolytic virus as an attractive purging strategy for autologous stem cell transplantations.
大剂量细胞毒性治疗后的血液学干细胞救援被广泛用于治疗多种血液系统和实体肿瘤。基因标记研究表明，自体移植物中的隐匿肿瘤细胞可能导致临床复发。迄今为止，清除被癌细胞污染的自体移植物一直未成功。呼肠孤病毒对体外、体内和离体系统中多种恶性组织学类型具有选择性溶瘤特性，此前已有相关证明。在本研究中，我们表明呼肠孤病毒能够成功清除自体移植物中的癌细胞。在实验性清除模型中，使用了人单核细胞和骨髓瘤细胞系以及富集的离体淋巴瘤、骨髓瘤和华氏巨球蛋白血症患者肿瘤标本。经呼肠孤病毒处理后，弥漫性大B细胞淋巴瘤、慢性淋巴细胞白血病、华氏巨球蛋白血症和小淋巴细胞淋巴瘤的细胞系或纯化的离体肿瘤细胞的活力显著降低。此外，细胞蛋白质的[35S]-甲硫氨酸标记和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）显示，早在24小时就有呼肠孤病毒蛋白质合成以及宿主细胞蛋白质合成的破坏。用呼肠孤病毒处理的上述肿瘤细胞和细胞系与单采产品的混合物显示疾病被完全清除。相比之下，呼肠孤病毒对富集的离体多发性骨髓瘤、伯基特淋巴瘤和滤泡性淋巴瘤的清除并不完全。即使在粒细胞集落刺激因子（G-CSF）刺激后，呼肠孤病毒的溶瘤作用也不影响CD34+干细胞或其长期集落形成试验。我们的结果表明，将未减毒的溶瘤病毒用于离体，是自体干细胞移植一种有吸引力的清除策略。
