Optimizing Viral Protein Yield of Influenza Virus Strain A/Vietnam/1203/2004 by Modification of the Neuraminidase Gene
Optimizing Viral Protein Yield of Influenza Virus Strain A/Vietnam/1203/2004 by Modification of the Neuraminidase Gene
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DOI：
10.1128/jvi.02391-08
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发表时间：
2009-05-01
影响因子：
5.4
通讯作者：
Ye, Zhiping
中科院分区：
文献类型：
--
作者：
Adamo, Joan E.;Liu, Teresa;Ye, Zhiping
高产流感病毒H5N1毒株的制备对研究人员和疫苗生产商都提出了挑战。 H5N1 毒株产量相对较低的原因尚不完全清楚，但可能部分取决于血凝素 (HA) 或神经氨酸酶 (NA) 表面糖蛋白与其他流感病毒蛋白之间的相互作用。在这项研究中，我们在 A/Puerto Rico/8/34 (PR8) NA 基因和 A/Vietnam/1203/2004 (VN1203) NA 基因之间构建了嵌合体，导致重配病毒的病毒产量增加，而 NA 活性没有显着损失。通过将 PR8 菌株的 NA 的氨基末端与 VN1203 的 NA 的羧基末端相结合，保留了 H5N1 VN1203 NA 糖蛋白特有的表面表位。这种重配病毒在卵中具有更高的滴度和总蛋白产量，生长至更高的滴度，在 MDCK 细胞上产生大噬斑，并保留 NA 活性。这项工作描述了一种新颖的重组技术，旨在提高生产大流行性流感病毒疫苗的候选疫苗的产量。还讨论了重配体的感染性和蛋白质产量之间的关系。
The preparation of high-yield prepandemic influenza virus H5N1 strains has presented a challenge to both researchers and vaccine manufacturers. The reasons for the relatively low yield of the H5N1 strains are not fully understood, but it might be partially dependent on the interactions between the hemagglutinin (HA) or neuraminidase (NA) surface glycoprotein and other influenza virus proteins. In this study, we have constructed chimeras between the A/Puerto Rico/8/34 (PR8) NA gene and the A/Vietnam/1203/2004 (VN1203) NA gene that have resulted in an increase in the virus yield of the reassortant viruses without a significant loss of NA activity. By combining the amino terminus of NA from the PR8 strain with the carboxy terminus of NA from VN1203, the surface epitopes unique to the H5N1 VN1203 NA glycoprotein are maintained. This reassortant virus had a higher titer and total protein yield in eggs, grew to a higher titer, produced large plaques on MDCK cells, and retained NA activity. This work describes a novel recombinant technique designed to increase the yields of vaccine candidates for the production of pandemic influenza virus vaccines. The relationship between the infectivity and protein yield of the reassortants also is discussed.