Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
PLoS One. 2013 Sep 4;8(9):e72530. doi: 10.1371/journal.pone.0072530. eCollection 2013.
Newcastle disease virus (NDV), an avian paramyxovirus type 1, is a promising vector for expression of heterologous proteins from a variety of unrelated viruses including highly pathogenic avian influenza virus (HPAIV). However, pre-existing NDV antibodies may impair vector virus replication, resulting in an inefficient immune response against the foreign antigen. A chimeric NDV-based vector with functional surface glycoproteins unrelated to NDV could overcome this problem. Therefore, an NDV vector was constructed which carries the fusion (F) and hemagglutinin-neuraminidase (HN) proteins of avian paramyxovirus type 8 (APMV-8) instead of the corresponding NDV proteins in an NDV backbone derived from the lentogenic NDV Clone 30 and a gene expressing HPAIV H5 inserted between the F and HN genes. After successful virus rescue by reverse genetics, the resulting chNDVFHN PMV8H5 was characterized in vitro and in vivo. Expression and virion incorporation of the heterologous proteins was verified by Western blot and electron microscopy. Replication of the newly generated recombinant virus was comparable to parental NDV in embryonated chicken eggs. Immunization with chNDVFHN PMV8H5 stimulated full protection against lethal HPAIV infection in chickens without as well as with maternally derived NDV antibodies. Thus, tailored NDV vector vaccines can be provided for use in the presence or absence of routine NDV vaccination.
新城疫病毒（NDV）是一种禽副黏病毒 1 型，是表达来自各种无关病毒（包括高致病性禽流感病毒（HPAIV））的异源蛋白的有前途的载体。然而，预先存在的 NDV 抗体可能会损害载体病毒的复制，导致对外来抗原的低效免疫反应。具有与 NDV 无关的功能性表面糖蛋白的嵌合 NDV 载体可以克服这个问题。因此，构建了一种 NDV 载体，该载体携带禽副黏病毒 8 型（APMV-8）的融合（F）和血凝素神经氨酸酶（HN）蛋白，而不是来自弱毒 NDV Clone 30 的 NDV 骨干中的相应 NDV 蛋白，以及在 F 和 HN 基因之间插入的表达 HPAIV H5 的基因。通过反向遗传学成功挽救病毒后，在体外和体内对产生的 chNDVFHN PMV8H5 进行了表征。通过 Western blot 和电子显微镜验证了异源蛋白的表达和病毒粒子的掺入。新生成的重组病毒的复制与在鸡胚中的亲本 NDV 相当。用 chNDVFHN PMV8H5 免疫可刺激鸡对致死性 HPAIV 感染的完全保护，而无需常规 NDV 疫苗接种或存在母源 NDV 抗体。因此，可以提供定制的 NDV 载体疫苗，用于存在或不存在常规 NDV 疫苗接种的情况。
