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Gene therapy for adult T-cell leukemia using HIV vector

使用 HIV 载体治疗成人 T 细胞白血病的基因治疗

基本信息

批准号：
07671218
负责人：
MATSUSHITA Shuzo
金额：
$ 1.54万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
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-07671218/
关键词：
gene therapy ATL HIV vector Thymidine kinase ganciclovir 成人T細胞白血病

项目摘要

Adult T-cell leukemia/lymphoma (ATL) is derived from CD4-positive T-cells, and has a poor prognosis because of its resistance to chemotherapy. To evaluate the effectiveness of gene therapy for ATL,the effect of ganciclovir on ATL cell lines transfected with the thymidine kinase gene of herpes simplex type 1 (HSV-TK) was analyzed. To tranfer the HSV-TK gene to ATL cells, an HIV-vector that has specific infectivity to CD4-positive cells was used. The thymidine kinase gene of herpes simplex virus type 1 was inserted into the long terminal repeats of human immunodeficiency virus type-1 (HIV-1) and driven by the SL3 promoter (named HXBSL3TK). HXBSL3TK was cotransfected with HXBCAT as a reporter into MT2 or HUT102 cells by DEAE-dextran. The cells were incubated with ganciclovir, and chloramphenicol acetyltransferase (CAT) activity was analyzed. The CAT activity of the MT2 cells and HUT102 cells transfected with HXBSL3TK decreased dosedependently with ganciclovir. HXBSL3TK was also contransfe … More cted into Cos cells with an HIV-1 packaging vector that has gag, pol and env driven by a cytomegalovirus promoter. The supernatant was transferred to MT2 cells or Raji cells, and incubated with ganciclovir. Ninety percent of the MT2 cells transduced by HXBSL3TK and incubated with ganciclovir were killed, but Raji cells were not killed. In addition HXBTK that expresses the HSV-TK gene and Tat gene driven by the LTR of HIV-1 was constructed. HXBTK had a higher expression of the HSV-TK gene and higher sensitivity to ganciclovir than did HXBSL3TK.Furthermore, the packaging cell lines were established, effective transduction to the fresh ATL cells were demonstrated.HIV vector has been shown to be one of the most promising new tools for human gene therapy strategy. In this study, the effect of ganciclovir on adult T-cell leukemia (ATL) cell lines transduced with HIV vector carrying the HSV-TK gene was evaluated. A high efficiency of killing of ATL cell lines and specificity for CD4-positive cells were demonstrated. These results suggest the effectiveness of HIV vector carrying the HSV-TK gene for gene therapy for ATL. Less

成人T细胞白血病/淋巴瘤（ATL）是由CD 4阳性T细胞衍生而来的，由于其对化疗的耐药性，预后不良。为了评价基因治疗ATL的有效性，分析了更昔洛韦对转染单纯疱疹1型胸苷激酶基因（HSV-TK）的ATL细胞系的作用。为了将HSV-TK基因转移至ATL细胞，使用了对CD 4阳性细胞具有特异性感染性的HIV载体。将单纯疱疹病毒1型胸苷激酶基因插入到人类免疫缺陷病毒1型（HIV-1）的长末端重复序列中，并由SL 3启动子驱动（命名为HXBSL 3 TK）。以HXBCAT为报告基因，用DEAE-dextran将HXBSL 3 TK共转染MT 2或HUT 102细胞。将细胞与更昔洛韦孵育，并分析氯霉素乙酰转移酶（CAT）活性。更昔洛韦可剂量依赖性地降低转染HXBSL 3 TK的MT 2细胞和HUT 102细胞的CAT活性。HXBSL 3 TK也被转基因。 ...更多信息用具有由巨细胞病毒启动子驱动的gag、pol和env的HIV-1包装载体导入Cos细胞。将上清液转移到MT 2细胞或Raji细胞中，并与更昔洛韦一起孵育。用HXBSL 3 TK转导的MT 2细胞与更昔洛韦孵育后，90%的MT 2细胞被杀死，但Raji细胞没有被杀死。此外，还构建了表达HSV-TK基因和由HIV-1的LTR驱动的达特基因的HXBTK。HXBTK基因表达量高，对更昔洛韦敏感性高，构建了包装细胞系，可有效转导新鲜ATL细胞，为人类基因治疗策略提供了新的工具。在这项研究中，更昔洛韦对成人T细胞白血病（ATL）细胞系转导携带HSV-TK基因的HIV载体的影响进行了评估。证明了对ATL细胞系的高杀伤效率和对CD 4阳性细胞的特异性。这些结果表明，携带HSV-TK基因的HIV载体用于ATL的基因治疗是有效的。少