Engineering viral vectors to subvert the airway defense response.
Engineering viral vectors to subvert the airway defense response.
复制标题
设计病毒载体来破坏气道防御反应。
DOI：
--
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发表时间：
1999
影响因子：
6.4
通讯作者：
S. Brody
中科院分区：
文献类型：
--
作者：
D. Look;S. Brody
通过将外源DNA递送至细胞以表达治疗性蛋白质来治疗肺部疾病的潜在治疗是分子工程的最近革命的令人兴奋的益处（1-3）。基于腺病毒的载体已经受到相当大的关注，用于将DNA递送到肺细胞，原因有以下几个：腺病毒可以感染气道上皮中的非分裂细胞;它们是稳定的;它们可以以高滴度产生;并且它们容易被修饰为复制缺陷型并表达外源基因。腺病毒载体已经在细胞培养系统（4，5）、动物模型（4，6，7）和人体试验（8-10）中证明了基因转移的能力。然而，随着这种治疗技术发展到体内系统，出现了基因治疗的生物学障碍，限制了高水平和延长的转基因表达（1-3）。宿主的抗病毒防御反应是一个障碍，特别是缓和了对早期腺病毒载体的热情，尽管最近对载体或宿主免疫的修饰在抑制这种反应方面取得了一些成功。因此，这个角度的目的是简要回顾宿主对腺病毒的反应，并讨论潜在的策略，以克服这一障碍，腺病毒为基础的基因治疗肺。将腺病毒载体施用到肺中产生复杂的多组分宿主应答，其涉及先天性和适应性免疫，导致对基因治疗功效的特异性抑制作用（2）。对腺病毒的肺部反应包括：（1）抗原非特异性、依赖于精氨酸的反应，导致急性炎症，可能具有神经原性成分并可引起全身毒性（7，9-12）;（2）针对表达病毒或转基因蛋白的细胞的主要组织相容性复合体（MHC）I类限制性细胞毒性（CD 81）T淋巴细胞依赖性应答，导致慢性炎症和缺乏持续的转基因表达（7，11，13）;和（3）针对载体递送期间存在的腺病毒衣壳蛋白的辅助（CD 4 1）T淋巴细胞依赖性应答，导致产生限制重复载体施用（如果需要维持治疗效果）的中和抗体（10，13，14）。对病毒病原体的反应是通过多种相互关联的机制发生的，这并不奇怪，因为多组分系统可能适应宿主对各种病原体的防御。然而，很明显，肺中抗病毒应答的所有组分都需要被鉴定和控制，以提高腺病毒载体的基因治疗功效。Thorne及其同事（15）提出了一种小鼠模型，在该模型中，高剂量暴露于灭活的腺病毒颗粒引起急性气道炎症反应，其特征是白细胞介素（IL）-6和肿瘤坏死因子α释放，随后中性粒细胞募集到肺中。该模型似乎与人类的基因治疗相关，因为在向囊性纤维化（CF）患者的肺施用腺病毒载体后观察到IL-6释放（9）。Thorne及其同事研究的对腺病毒的早期肺反应似乎与对内毒素的反应无关，这表明两种刺激可能通过重叠途径激活炎症。这一发现对患有内毒素预先存在的炎症的患者的基于腺病毒的基因治疗具有意义，因为这是患有肺部疾病如CF的患者的气道环境的一部分，其特征在于革兰氏阴性杆菌的慢性感染。该研究还证实，宿主对腺病毒的部分应答不依赖于载体基因表达，但可能受到调节（16）。由于这种急性反应似乎是独立的病毒基因表达，未来的工作将需要针对识别的启动元件在腺病毒颗粒的希望，这些元素可能会被改变，以限制宿主的反应。由于宿主对腺病毒载体的反应限制了基于腺病毒的基因转移的效用，因此已经测试了两种主要策略来减弱这些反应。第一种策略是在鉴定启动或放大肺防御反应的特定病毒分子的基础上形成的，随后进行载体再工程改造以删除它们的表达。这一方向的工作跨越了从去除特定的病毒基因产物（例如，早期区域2A）至完全去除表达病毒蛋白的载体中的所有基因（17，18）。第二种策略是使用药物（例如，皮质类固醇，环磷酰胺），介质（例如，IL-12）或其他阻断策略（例如，抗细胞受体蛋白，口服耐受化）（19-22）。两者（1999年4月2日收到原件）
The potential treatment of lung disease by delivery of exogenous DNA to cells for expression of therapeutic proteins is an exciting benefit of the recent revolution in molecular engineering (1–3). Adenovirus-based vectors have received considerable attention for DNA delivery to lung cells for several reasons: adenoviruses can infect nondividing cells in the airway epithelium; they are stable; they can be produced in high titer; and they are easily modified to be replication-deficient and express exogenous genes. Adenoviral vectors have already demonstrated the capacity for gene transfer in cell culture systems (4, 5), animal models (4, 6, 7), and human trials (8–10). However, as development of this therapeutic technology progressed to in vivo systems, biologic obstacles to gene therapy have emerged that limit high-level and prolonged transgene expression (1–3). The antiviral defense response of the host is one obstacle that has particularly tempered enthusiasm for earlygeneration adenoviral vectors, although recent modifications of vectors or host immunity have had some success at inhibiting this response. Accordingly, the purpose of this perspective is to review briefly the host response to adenoviruses and to discuss potential strategies to overcome this obstacle to adenovirus-based gene therapy to the lung. Administration of adenoviral vectors into the lung generates a complex, multicomponent host response that involves innate and adaptive immunity, resulting in specific inhibitory effects on gene therapy efficacy (2). Components of the pulmonary response to adenovirus include: ( 1 ) an antigen-nonspecific, cytokine-dependent response resulting in acute inflammation, which may have a neurogenic component and can cause systemic toxicity (7, 9–12); ( 2 ) a major histocompatibility complex (MHC) class I– restricted, cytotoxic (CD8 1 ) T lymphocyte–dependent response directed at cells expressing viral or transgene proteins, resulting in chronic inflammation and a lack of persistent transgene expression (7, 11, 13); and ( 3 ) a helper (CD4 1 ) T lymphocyte–dependent response directed at adenoviral capsid proteins present during vector delivery, resulting in the production of neutralizing antibodies that limit repeated vector administration (if required to sustain a therapeutic effect) (10, 13, 14). It should not be surprising that the response to viral pathogens occurs through multiple, interrelated mechanisms because a multicomponent system can probably adapt for host-defense against a variety of pathogens. However, it is apparent that all components of the antiviral response in the lung will need to be identified and controlled in order to improve gene therapy efficacy with adenoviral vectors. In this issue, Thorne and colleagues (15) present a murine model in which high-dose exposure to inactivated adenoviral particles elicits an acute airway inflammatory response marked by interleukin (IL)-6 and tumor necrosis factora release and consequent neutrophil recruitment into the lung. This model appears relevant to gene therapy in humans because IL-6 release was observed after adenoviral vector administration to the lungs of patients with cystic fibrosis (CF) (9). The early pulmonary response to the adenovirus studied by Thorne and colleagues does not appear to be additive with the response to endotoxin, suggesting that both stimuli may activate inflammation by overlapping pathways. This finding has implications for adenovirus-based gene therapy in patients with preexisting inflammation from endotoxin, as this is part of the airway milieu in patients with lung diseases such as CF, characterized by chronic infection with gram-negative bacilli. This study also confirms that a portion of the host response to adenovirus is independent of vector gene expression, but may be modulated pharmacologically (16). Because this acute response appears to be independent of viral gene expression, future work will need to be directed toward identification of the initiating elements in adenoviral particles with the hope that these elements might be altered to limit the host response. As host responses to adenoviral vectors that limit the utility of adenovirus-based gene transfer have been identified, two major strategies to blunt these responses have been tested. The first strategy is formed on the basis of identifying specific viral molecules that initiate or amplify the pulmonary defense response with subsequent vector reengineering to delete their expression. Work in this direction spans from removal of specific viral gene products (e.g., early region 2A) to total removal of all genes in the vectors that express viral proteins (17, 18). A second strategy is to pharmacologically manipulate the host response using drugs (e.g., corticosteroids, cyclophosphamide), mediators (e.g., IL-12), or other blocking strategies (e.g., anticell receptor proteins, oral tolerization) (19–22). Both ( Received in original form April 2, 1999 )
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影响因子：
158.5
作者：
KNOWLES, MR;HOHNEKER, KW;BOUCHER, RC
通讯作者：
BOUCHER, RC
E2a 缺陷的重组腺病毒在棉鼠肺中延长转基因表达。
影响因子：
4.2
作者：
Engelhardt,JF;Litzky,L;Wilson,JM
通讯作者：
Wilson,JM
影响因子：
56.9
作者：
Darnell, JE
通讯作者：
Darnell, JE
对腺病毒抗原的口服耐受允许使用重组腺病毒载体进行长期基因表达。
DOI：
10.1172/jci119238
发表时间：
1997
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Ilan,Y;Prakash,R;Davidson,A;Jona;Droguett,G;Horwitz,MS;Chowdhury,NR;Chowdhury,JR
通讯作者：
Chowdhury,JR
体内腺病毒介导的小鼠肝细胞转基因表达的菌株相关变异：免疫活性和免疫缺陷近交菌株之间的比较。
DOI：
--
发表时间：
1995
期刊：
Gene therapy.
影响因子：
--
作者：
Barr,D;Tubb,J;Ferguson,D;Scaria,A;Lieber,A;Wilson,C;Perkins,J;Kay,MA
通讯作者：
Kay,MA