Retroviral immunotoxin gene therapy of leukemia in mice using leukemia-specific T cells transduced with an interleukin-3/Bax fusion protein gene.
Retroviral immunotoxin gene therapy of leukemia in mice using leukemia-specific T cells transduced with an interleukin-3/Bax fusion protein gene.
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使用转染白细胞介素 3/Bax 融合蛋白基因的白血病特异性 T 细胞对小鼠白血病进行逆转录病毒免疫毒素基因治疗。
DOI：
10.1089/104303403322611791
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Chen,Wei
中科院分区：
文献类型：
--
作者：
Vallera,DanielA;Jin,Ni;Shu,Yanqun;Panoskaltsis-Mortari,Angela;Kelekar,Ameeta;Chen,Wei
在过去的研究中，我们表明，T细胞转导与逆转录病毒白喉免疫毒素（IT）的靶基因可以作为车辆提供IT到体内肿瘤。我们利用了 抗原特异性T细胞能够穿透肿瘤，以设计一种将细胞和体液治疗直接递送到肿瘤部位的方法。为了提高肿瘤特异性，我们选择白细胞介素 （IL）-3作为配体，因为其受体在髓系白血病祖细胞上选择性过表达。由于Bcl-2家族蛋白与白喉毒素（DT）显示出结构相似性，我们构建了一个 独特的逆转录病毒IT使用Bax，Bcl-2家族的促凋亡成员，代替DT。选择Bax是因为几项研究表明，它的转导在不同的癌症中诱导致死性细胞凋亡。 用于基因治疗的逆转录病毒构建体包括位于其80个氨基酸前导序列下游的IL-3，并且允许杂合IL-3/人Bax融合蛋白的共翻译蛋白合成。其他 用IL-3与DT或假单胞菌外毒素融合构建载体。逆转录病毒载体用于瞬时转染C8，一种特异性识别FBL-3的CD 4 +T细胞克隆， 一种致命的骨髓性白血病从转导细胞收集的上清液显示出促凋亡活性，并在体外选择性地抑制FBL-3细胞。腹膜内注射转导的但不 未转导的C8进入具有皮下肿瘤或全身性癌症的小鼠显著抑制肿瘤生长。这些结果表明，用IL-3和各种毒性蛋白制备的逆转录病毒IT可能是有用的 急性髓性白血病（AML）患者的治疗。此外，Bax构建体可以特别用作retIT中细菌毒素的非免疫原性替代物。
In past studies, we showed that T cells transduced with retroviral diphtheria immunotoxin (IT) target genes could serve as vehicles for delivering IT to tumorsin vivo. We took advantage of the observation that antigen-specific T cells are able to penetrate tumors to design an approach delivering combined cellular and humoral therapy directly to the tumor site. To improve tumor specificity, we selected interleukin (IL)-3 as a ligand because its receptor is selectively overexpressed on myeloid leukemia progenitors. Because Bcl-2 family proteins show structural similarity to diphtheria toxin (DT), we constructed a unique retroviral IT using Bax, a proapoptotic member of the Bcl-2 family, in place of DT. Bax was chosen because several studies showed that its transduction induces lethal apoptosis in different cancers. The retroviral construct for gene therapy included IL-3 positioned downstream of its 80 amino acid leader, and permitted cotranslational protein synthesis of hybrid IL-3/human Bax fusion protein. Other vectors were constructed with IL-3 fused to DT orPseudomonasexotoxin. Retroviral vectors were used to transiently transduce C8, a CD4+T cell clone that specifically recognized FBL-3, a lethal myeloid leukemia. Supernatants collected from transduced cells showed proapoptotic activity and selectively inhibited FBL-3 cellsin vitro. Intraperitoneal injection of transduced but not nontransduced C8 into mice with subcutaneous tumors or systemic cancer significantly inhibited tumor growth. These results indicate that retroviral IT made with IL-3 and various toxic proteins may be useful in patients with acute myelogenous leukemia (AML). Furthermore, the Bax construct may be particularly useful as a nonimmunogenic substitute for bacterial toxins in retIT.