Ramp Kristina, Veits Jutta, Deckers Daniela, Rudolf Miriam, Grund Christian, Mettenleiter Thomas C, Römer-Oberdörfer Angela
Institute of Molecular Biology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Südufer 10, D-17493 Greifswald-Insel Riems, Germany.
Avian Dis. 2011 Sep;55(3):413-21. doi: 10.1637/9652-011111-Reg.1.
To analyze the contribution of neuraminidase (NA) toward protection against avian influenza virus (AIV) infection, three different recombinant Newcastle disease viruses (NDVs) expressing hemagglutinin (HA) or NA, or both, of highly pathogenic avian influenza virus (HPAIV) were generated. The lentogenic NDV Clone 30 was used as backbone for the insertion of HA of HPAIV strain A/chicken/Vietnam/P41/05 (H5N1) and NA of HPAIV strain A/duck/Vietnam/TG24-01/05 (H5N1). The HA was inserted between the genes encoding NDV phosphoprotein (P) and matrixprotein (M), and the NA was inserted between the fusion (F) and hemagglutinin-neuraminidase protein (HN) genes, resulting in NDVH5VmPMN1FHN. Two additional recombinants were constructed carrying the HA gene between the NDV P and M genes (NDVH5VmPM) or the NA between F and HN (NDVN1FHN). All recombinants replicated well and stably expressed the HA gene, the NA gene, or both. Chickens immunized with NDVH5VmPMN1FHN or NDVH5VmPM were protected against two different HPAIV H5N1 and also against HPAIV H5N2. In contrast, immunization of chickens with NDVN1FHN induced NDV- and AIV N1-specific antibodies but did not protect the animals against a lethal dose of HPAIV H5N1. Furthermore, expression of AIV N1, in addition to AIV H5 by NDV, did not increase protection against HPAIV H5N1.
为分析神经氨酸酶（NA）在预防禽流感病毒（AIV）感染中的作用，构建了三种不同的重组新城疫病毒（NDV），分别表达高致病性禽流感病毒（HPAIV）的血凝素（HA）或NA，或两者都表达。弱毒NDV克隆30用作骨架，插入HPAIV毒株A/鸡/越南/P41/05（H5N1）的HA和HPAIV毒株A/鸭/越南/TG24 - 01/05（H5N1）的NA。HA插入到编码NDV磷蛋白（P）和基质蛋白（M）的基因之间，NA插入到融合蛋白（F）和血凝素 - 神经氨酸酶蛋白（HN）基因之间，得到NDVH5VmPMN1FHN。另外构建了两个重组体，一个在NDV的P和M基因之间携带HA基因（NDVH5VmPM），另一个在F和HN之间携带NA基因（NDVN1FHN）。所有重组体均能良好复制并稳定表达HA基因、NA基因或两者。用NDVH5VmPMN1FHN或NDVH5VmPM免疫的鸡对两种不同的HPAIV H5N1以及HPAIV H5N2具有抵抗力。相比之下，用NDVN1FHN免疫鸡可诱导产生NDV和AIV N1特异性抗体，但不能保护动物免受致死剂量的HPAIV H5N1感染。此外，NDV除表达AIV H5外还表达AIV N1，并未增强对HPAIV H5N1的抵抗力。
