Influenza M2 envelope protein augments avian influenza hemagglutinin pseudotyping of lentiviral vectors
Influenza M2 envelope protein augments avian influenza hemagglutinin pseudotyping of lentiviral vectors
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DOI：
10.1038/sj.gt.3302715
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发表时间：
2006-04-01
期刊：
影响因子：
5.1
通讯作者：
Olsen, JC
中科院分区：
文献类型：
--
作者：
McKay, T;Patel, M;Olsen, JC
基于慢病毒的基因转移有可能有效地将基于dna的疗法传递到气道的非分裂上皮细胞中，用于治疗肺部疾病，如囊性纤维化。然而，在将慢病毒载体常规应用于肺部之前，必须克服肺特异性基因转移和慢病毒载体生产的重大障碍。在本研究中，我们研究了流感病毒M2在载体生成细胞中的共表达是否可以提高禽鼠疫病毒血凝素（HA）伪型慢病毒载体的生成能力。我们发现M2的表达导致基于马传染性贫血病毒（EIAV）或人类免疫缺陷病毒1型（HIV-1）的ha假型慢病毒载体的产量增加10-30倍。利用M2抑制剂金刚烷胺和M2耐药突变体进行的实验表明，M2的离子通道活性对M2依赖性载体产量的增加很重要。此外，从产生细胞释放颗粒所需的神经氨酸酶活性也可以通过流感NA cDNA的共表达纳入产生细胞。以流感包膜蛋白为假型的慢病毒载体能够有效地在体外通过极化小鼠气管培养物的顶膜以及在体内小鼠气管上皮进行转导。
Lentivirus-based gene transfer has the potential to efficiently deliver DNA-based therapies into non-dividing epithelial cells of the airway for the treatment of lung diseases such as cystic fibrosis. However, significant barriers both to lung-specific gene transfer and to production of lentivirus vectors must be overcome before these vectors can be routinely used for applications to the lung. In this study, we investigated whether the ability to produce lentiviral vectors pseudotyped with fowl plague virus hemagglutinin (HA) could be improved by co-expression of influenza virus M2 in vector-producing cells. We found that M2 expression led to a 10-30-fold increase in production of HA-pseudotyped lentivirus vectors based upon equine infectious anemia virus (EIAV) or human immunodeficiency virus type 1 (HIV-1). Experiments using the M2 inhibitor amantadine and a drug-resistant mutant of M2 established that the ion channel activity of M2 was important for M2-dependent augmentation of vector production. Furthermore, the neuraminidase activity necessary for particle release from producer cells could also be incorporated into producer cells by co-expression of influenza NA cDNA. Lentiviral vectors pseudotyped with influenza envelope proteins were able to efficiently transduce via the apical membrane of polarized mouse tracheal cultures in vitro as well as mouse tracheal epithelia in vivo.