Jinoch Pavel, Zak Roman, Janousková Olga, Kunke David, Rittich Simon, Duskova Martina, Sobotkova Eva, Marinov Iuri, Andelova Monika, Smahel Michal, Vonka Vladimir
Department of Experimental Virology, Institute of Hematology and Blood Transfusion, U Nemocnice 1, 128-20 Prague 2, Czech Republic.
Int J Oncol. 2003 Sep;23(3):775-83.
From mouse (C57BL/6) HPV-16 transformed cells denoted MK16/1/IIIABC (MK16) a cellular thymidine kinase deficient (cTK-) cell line was isolated. These cTK- cells were transduced by bicistronic recombinant adeno-associated viruses (rAAV) carrying the herpes simplex virus thymidine kinase gene and the gene for either the mouse granulocyte-macrophage colony stimulating factor (GM-CSF) or mouse interleukin-2 (IL-2). Transduced cells were highly sensitive to minute amounts of ganciclovir (GCV) and synthesized moderate amounts of the respective cytokines. A number of cell clones were tested for the cytokine production. The two best producer cell lines, the GM-CSF-producing cells denoted B9 and the IL-2-producing cells denoted 181, were selected for further experiments. Neither B9 nor 181 cells were tumorigenic in syngeneic animals. As inducers of antitumour immunity against challenge with MK16 cells, B9 cells proved superior to the 181 cells. GCV treatment did not markedly influence the level of immunity induced.
从标记为MK16/1/IIIABC（MK16）的小鼠（C57BL/6）人乳头瘤病毒16型转化细胞中分离出一种细胞胸苷激酶缺陷（cTK-）细胞系。这些cTK-细胞用携带单纯疱疹病毒胸苷激酶基因以及小鼠粒细胞-巨噬细胞集落刺激因子（GM-CSF）基因或小鼠白细胞介素2（IL-2）基因的双顺反子重组腺相关病毒（rAAV）进行转导。转导后的细胞对微量更昔洛韦（GCV）高度敏感，并合成适量的相应细胞因子。对多个细胞克隆进行了细胞因子产生情况的检测。选择了两个最佳的产生细胞系，即产生GM-CSF的标记为B9的细胞和产生IL-2的标记为181的细胞用于进一步实验。B9细胞和181细胞在同基因动物中均无致瘤性。作为针对MK16细胞攻击的抗肿瘤免疫诱导剂，B9细胞被证明优于181细胞。GCV处理并未显著影响诱导的免疫水平。
