Gilligan M G, Knox P, Weedon S, Barton R, Kerr D J, Searle P, Young L S
CRC Institute for Cancer Studies, University of Birmingham, Edgbaston, UK.
Gene Ther. 1998 Jul;5(7):965-74. doi: 10.1038/sj.gt.3300672.
The majority of tumour cells do not express immune costimulatory molecules and this may account for their inability to stimulate directly an antitumour T cell response. Here we report on the construction of a recombinant E1/E3-deleted adenovirus encoding the human B7-1 costimulatory molecule. We explored the use of this vector for gene transfer to a number of human ovarian and cervical tumour cell lines, and to primary ovarian tumour material. Rapid and efficient gene transfer and expression was obtained in the majority of cases using a multiplicity of infection of 30 plaque forming units per cell. B7-1 expression was detectable at the cell surface within 12 h and was still detectable 10 days after infection. The immunogenicity of gene-modified tumour cells was tested in an allogeneic mixed lymphocyte tumour cell culture. Tumour cells expressing B7-1 were found to induce significantly higher levels of T cell proliferation than tumour cells modified with a control adenovirus carrying the beta-galactosidase gene. B7-1-induced T cell proliferation could be blocked by the addition of anti-B7-1 antibodies at the initiation of cocultures. These results support the rationale for use of adenovirally delivered B7-1 for genetic immunotherapy of ovarian and cervical cancer.
大多数肿瘤细胞不表达免疫共刺激分子，这可能是它们无法直接刺激抗肿瘤T细胞反应的原因。在此，我们报告了一种编码人B7-1共刺激分子的重组E1/E3缺失腺病毒的构建。我们探索了使用该载体将基因转移到多种人卵巢和宫颈肿瘤细胞系以及原发性卵巢肿瘤材料中的方法。在大多数情况下，使用每细胞30个噬斑形成单位的感染复数可实现快速高效的基因转移和表达。感染后12小时内即可在细胞表面检测到B7-1表达，感染10天后仍可检测到。在同种异体混合淋巴细胞肿瘤细胞培养中测试了基因修饰肿瘤细胞的免疫原性。发现表达B7-1的肿瘤细胞比用携带β-半乳糖苷酶基因的对照腺病毒修饰的肿瘤细胞诱导更高水平的T细胞增殖。在共培养开始时添加抗B7-1抗体可阻断B7-1诱导的T细胞增殖。这些结果支持了使用腺病毒递送的B7-1进行卵巢癌和宫颈癌基因免疫治疗的理论依据。
