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Induction of tumor specific cytotoxic T lymphocyte by using transfer of co-simulatory molecule gene

共模拟分子基因转移诱导肿瘤特异性细胞毒性T淋巴细胞

基本信息

批准号：
07457150
负责人：
SETOGUCHI Yasuhiro
金额：
$ 4.35万
依托单位：
Juntendo University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457150/
关键词：
gene trasfer adenovirus vector co-stimulatory molecule CD80 CD86 cytotoxic T lymphocyte cancer 癌 B7遺伝子遺伝子治療腫瘍分子免疫

项目摘要

Interaction between the costimulatory molecule B7-1 (CD80) on antigen-presenting cells and its counter-receptor CD28 on T lymphocyte plays a key role in the induction of cell-mediated immune responses including cancer. Many solid tumor lack expression of B7-1 and this has been suggested to contribute to the failure of immune recognition of these diseases. Based on this knowledge, we hypothesized that co-stimulatory signal delivered through B7-1 or B7-2 molecule expressed on lung cancer cells using replication deficient adenovirus vector (Ad) would induce tumor-specific cytotoxic T lymphocyte (CTL). To evaluate this hypothesis, we constructed two Ads ; AdCMVhB7 (am E1^- Ad5 based vector containing human B7-1 cDNA driven by cytomegalovirus immediate early promoter and enhancer) ; AdNull (above same vector withour expression of exogenous gene) as control. Theoretical advantages of Ad consist in capacity of high efficient gene transduction. Using these Ads, generation of tumor specific CTL … More was studied in a primary syngeneic mixed lymphocyte tumor culture by using both lung cancer cells and peripheral blood lymphocytes obtained from patients with lung cancer. Inoculation of lung cancer cells with 10 multiplicity of infection of AdCMVhB7 resulted in rapid and efficient cell surface expression of B7-1 molecule (>90% of cell at 24h). Cytolytic activity of lymphocytes by using ^<51>Cr-releasing assay (E/T=40) demonstrated that effector lymphocytes induced by hB7-1(+)lung cancer cells treated with AdCMVhB7 could lyse 45% parental lung cancer cells hB7-1(-) with a maximum lysis of 62%, in addition, the effector lymphocytes could not lyse the irrelevant syngeneic fibroblast. In countrast, effector lymphocytes induced by lung cancer cells treated with AdNull as control virus or PBS as control could not lyse parental lung cancer cells at all. Furthermore, cytolytic activity of the effector lymphocytes induced by B7-1 transduced lung cancer cells was inhibited by addition of anti-CD3 antibody (10ug/ml). These data suggested that effector lymphocytes induced by lung cancer treated with AdCMVhB7 have a character of tumor-specific CTL.Adenovirus mediated-hB7-1 gene transfer may be a useful means for gene therapy for lung cancer in application of adoptive immunotherapy. Less

抗原提呈细胞表面的共刺激分子B7-1（CD 80）与T淋巴细胞表面的CD 28相互作用在包括肿瘤在内的细胞免疫应答中起着关键作用。许多实体瘤缺乏B7-1的表达，这被认为是导致这些疾病的免疫识别失败的原因。在此基础上，我们假设通过复制缺陷型腺病毒载体（Ad）表达的B7-1或B7-2分子传递共刺激信号，可以诱导肿瘤特异性细胞毒性T淋巴细胞（CTL）。为了验证这一假设，我们构建了两个Ads：AdCMVhB 7（一个基于E1^-Ad 5的载体，含有由巨细胞病毒立即早期启动子和增强子驱动的人B7-1 cDNA）; Ad的理论优势在于其高效的基因转导能力。利用这些Ads，产生肿瘤特异性CTL， ...更多信息通过使用肺癌细胞和从肺癌患者获得的外周血淋巴细胞，在原代同基因混合淋巴细胞肿瘤培养物中研究。用10倍的AdCMVhB 7感染肺癌细胞，导致B7-1分子在细胞表面的快速和有效表达（在24小时>90%的细胞）。淋巴细胞溶<51>铬实验（E/T=40）表明，经AdCMVhB 7处理的hB 7 -1（+）肺癌细胞所诱导的效应淋巴细胞对hB 7 -1（-）亲本肺癌细胞的杀伤率为45%，最高达62%，且不能杀伤无关的同源成纤维细胞。相反，以Adhesive为对照病毒或PBS为对照的肺癌细胞诱导的效应淋巴细胞不能裂解亲本肺癌细胞。此外，抗CD 3抗体（10 μ g/ml）可抑制B7-1转导的肺癌细胞诱导的效应淋巴细胞的杀伤活性。结果表明，AdCMV-hB 7诱导的肺癌效应淋巴细胞具有肿瘤特异性CTL的特性，AdCMV-hB 7 -1基因转移有望成为过继免疫治疗肺癌的一种有效手段。少