Loss of transgene following ex vivo gene transfer is associated with a dominant Th2 response:: Implications for cutaneous gene therapy
Loss of transgene following ex vivo gene transfer is associated with a dominant Th2 response:: Implications for cutaneous gene therapy
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DOI：
10.1038/mt.sj.6300086
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发表时间：
2007-05-01
影响因子：
12.4
通讯作者：
Ghazizadeh, Soosan
中科院分区：
文献类型：
--
作者：
Lu, Zhenmei;Ghazizadeh, Soosan
宿主对治疗性基因产品的反应是皮肤基因治疗中潜在的严重并发症。因此，控制对治疗性抗原的免疫反应可能是有效治疗的关键。体外和体内向表皮干细胞的基因转移都能诱导转基因特异性免疫反应，但其免疫激活机制是否相同尚不清楚。在这项研究中，我们研究了以绿色荧光蛋白为模型抗原和逆转录病毒介导的基因传递的表皮基因转移的体外模型中转基因特异性免疫反应。体外转导的角质形成细胞的排斥反应与Th2/嗜酸性粒细胞反应相关，而与体内基因转导到表皮后引起的T辅助(Th)1型反应相反。这些反应的特征是T细胞产生IL-4和IL-5，血清中以抗绿色荧光蛋白IgG1为主，排斥皮肤中存在大量嗜酸性粒细胞，缺乏I类限制性细胞毒性T淋巴细胞反应。用中和抗IL-5抗体对接受体外转导的角质形成细胞的小鼠进行预处理，可以防止嗜酸性粒细胞的渗透，延长转导的表皮的存活时间。这些数据表明Th2/嗜酸性粒细胞通路在体外转导的角质形成细胞的排斥反应中发挥了作用，这表明了对体外和体内基因治疗方法实现耐受的不同要求。
Host responses to therapeutic gene products are potentially serious complications in cutaneous gene therapy. Controlling immune responses to the therapeutic antigen may therefore be critical for an effective therapy. Both ex vivo and in vivo gene transfer to epidermal stem cells has been shown to induce transgene-specific immune responses; however, whether the mechanism of immune activation is the same is not clear. In this study, we have characterized transgene-specific immune responses in an ex vivo model of epidermal gene transfer using green fluorescent protein as a model antigen and retrovirus-mediated gene delivery. Contrary to T helper (Th)1-type responses induced following in vivo gene transfer to epidermis, rejection of ex vivo-transduced keratinocytes was associated with Th2/eosinophilc responses. These responses were characterized by interleukin (IL)-4 and IL-5 production by T cells, a predominance of anti-green fluorescent protein IgG1 in serum, the presence of numerous eosinophils within rejected skin, and a lack of class I-restricted cytotoxic T lymphocyte response. Pretreatment of mice receiving ex vivo transduced keratinocytes with neutralizing anti-IL-5 antibody prevented eosinophil infiltration and prolonged survival of transduced epidermis. These data indicate a role for the Th2/eosinophilic pathway in rejection of ex vivo-transduced keratinocytes, suggesting different requirements for achieving tolerance for ex vivo and in vivo approaches to gene therapy.