Host immune responses in ex vivo approaches to cutaneous gene therapy targeted to keratinocytes
Host immune responses in ex vivo approaches to cutaneous gene therapy targeted to keratinocytes
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DOI：
10.1111/j.1600-0625.2005.00351.x
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发表时间：
2005-10-01
影响因子：
3.6
通讯作者：
Ghazizadeh, S
中科院分区：
文献类型：
--
作者：
Lu, Z;Ghazizadeh, S
表皮基因治疗可能有益于各种遗传性皮肤病和某些系统性疾病。体内和离体的基因转移方法都已被用于靶向人表皮干细胞，并在免疫缺陷小鼠/人嵌合体模型中实现长期转基因表达。然而，免疫应答，特别是在表达新抗原的情况下，可能会减少表达，从而限制治疗。体内基因转移到皮肤已经显示出诱导转基因特异性免疫应答。然而，离体基因转移方法，其中角质形成细胞在培养物中转导并移植回患者，可避免通过体内施用载体向免疫系统提供信号。在目前的研究中，我们已经开发了一个稳定的表皮移植平台，在免疫功能正常的小鼠中分析宿主在离体表皮基因治疗中的反应。我们使用绿色荧光蛋白（GFP）作为新抗原，并通过离体逆转录病毒介导的基因转移到耗尽抗原递呈细胞（APC）的小鼠原代表皮培养物中，显示出GFP特异性免疫反应的诱导，从而导致转导细胞的清除。在对GFP耐受的免疫活性小鼠中的类似方法导致转导细胞的永久植入和持续的GFP表达。在靶向角质形成细胞的离体基因转移中，转基因特异性免疫应答的激活需要转基因产物向APC的交叉呈递，这是一个最适合免疫调节的过程。该模型可用于探索将转基因特异性免疫应答转移到破坏性较小或致耐受性较低的免疫应答的策略。
Epidermal gene therapy may benefit a variety of inherited skin disorders and certain systemic diseases. Both in vivo and ex vivo approaches of gene transfer have been used to target human epidermal stem cells and achieve long-term transgene expression in immunodeficient mouse/human chimera models. Immunological responses however, especially in situations where a neoantigen is expressed, are likely to curtail expression and thereby limit the therapy. In vivo gene transfer to skin has been shown to induce transgene-specific immune responses. Ex vivo gene transfer approaches, where keratinocytes are transduced in culture and transplanted back to patient, however, may avoid signals provided to the immune system by in vivo administration of vectors. In the current study, we have developed a stable epidermal graft platform in immunocompetent mice to analyze host responses in ex vivo epidermal gene therapy. Using green fluorescent protein (GFP) as a neoantigen and an ex vivo retrovirus-mediated gene transfer to mouse primary epidermal cultures depleted of antigen-presenting cells (APCs), we show induction of GFP-specific immune responses leading to the clearance of transduced cells. Similar approach in immunocompetent mice tolerant to GFP resulted in permanent engraftment of transduced cells and continued GFP expression. Activation of transgene-specific immune responses in ex vivo gene transfer targeted to keratinocytes require cross-presentation of transgene product to APCs, a process that is most amenable to immune modulation. This model may be used to explore strategies to divert transgene-specific immune responses to less destructive or tolerogenic ones.