Retroviral transduction of human peripheral blood lymphocytes with bcl-xL promotes in vitro lymphocyte survival in pro-apoptotic conditions
Retroviral transduction of human peripheral blood lymphocytes with bcl-xL promotes in vitro lymphocyte survival in pro-apoptotic conditions
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DOI：
10.1038/sj.gt.3301685
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发表时间：
2002-04-01
期刊：
影响因子：
5.1
通讯作者：
Hawkins, RE
中科院分区：
文献类型：
--
作者：
Eaton, D;Gilham, DE;Hawkins, RE
基因修饰效应细胞在体内的长期存活对于任何（基因修饰的）过继细胞免疫治疗方法的成功至关重要。然而，在迄今为止的癌症临床试验中，检测存活的循环基因修饰T细胞需要高度敏感的技术。T细胞共刺激的体外研究表明，通过连接CD28上调抗凋亡基因Bcl-X-L可促进T细胞存活，但不能促进增殖。在这里，我们研究了通过使用表达Bcl-X-L的逆转录病毒载体转导人外周血淋巴细胞来调节细胞凋亡抗性和提高细胞存活率的能力。我们发现用Bcl-X-L逆转录病毒转导的Jurkat细胞对Fas （CD95）抗体诱导的细胞凋亡具有部分抗性。随后用转导的原代人淋巴细胞进行的体外实验表明，Bcl-XL的过表达促进了在缺乏白细胞介素-2的情况下培养的淋巴细胞的存活。通过抗cd3 （epsilon）抗体激活诱导的细胞凋亡，OKT3也被调节。此外，Bcl-X-L在人淋巴细胞中的过表达延缓了与肿瘤细胞系长期共培养诱导的细胞凋亡的发生。尽管体外存活率有所提高，但在安全性评估的初步实验中，在注射Bc1-X-L转导淋巴细胞的NOD/SCID小鼠中未观察到恶性肿瘤或自身免疫的迹象。这些结果表明，在淋巴细胞治疗中单独表达Bcl-X-L或与另一个治疗基因联合表达Bcl-X-L可以增强细胞在体内的持久性，从而可能改善过继细胞治疗的临床结果。
The prolonged in vivo survival of genetically modified effector cells is crucial to the success of any (gene-modified) adoptive cellular immunotherapy approach. In cancer clinical trials to date, however, the detection of surviving circulating gene-modified T cells has required highly sensitive techniques. In vitro studies of T cell co-stimulation have shown that up-regulation of the anti-apoptosis gene Bcl-X-L by ligation of CD28 promotes T cell survival, but not proliferation. Here we have investigated the ability to modulate resistance to apoptosis and improve cell survival by transducing human peripheral blood lymphocytes using a retroviral vector that expresses Bcl-X-L. We show that Jurkat cells transduced with Bcl-X-L retrovirus were partially resistant to Fas (CD95) antibody-induced apoptosis. Subsequent in vitro assays with transduced primary human lymphocytes demonstrates that over-expression of Bcl-XL promotes the survival of lymphocytes cultured in the absence of interleukin-2. Activation-induced apoptosis with anti-CD3(epsilon) antibody, OKT3 is also modulated. Furthermore, Bcl-X-L over-expression In human lymphocytes delays the onset of apoptosis induced by long-term co-culture with tumour cell lines. Despite this improved in vitro survival, in a preliminary experiment to assess safety, no signs of malignancy or autoimmunity were observed in NOD/SCID mice injected with Bc1-X-L transduced lymphocytes. These results indicate that expression of Bcl-X-L in lymphocyte therapy either alone or in conjunction with an additional therapeutic gene could enhance persistence of cells in vivo thereby potentially improving the clinical outcome of adoptive cellular therapy.