Recombinant human metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: Deletion of G yields a promising vaccine candidate
Recombinant human metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: Deletion of G yields a promising vaccine candidate
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DOI：
10.1128/jvi.78.23.12877-12887.2004
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发表时间：
2004-12-01
影响因子：
5.4
通讯作者：
Buchholz, UJ
中科院分区：
文献类型：
--
作者：
Biacchesi, S;Skiadopoulos, MH;Buchholz, UJ
人偏肺病毒（HMPV）是近年来发现的引起人类严重呼吸道疾病的重要病原体。特别是，关于HMPV对儿科呼吸道疾病的贡献的新信息表明，开发针对该病毒的疫苗与针对人呼吸道合胞病毒和人副流感病毒开发的疫苗联合使用将是重要的。最近描述的反向遗传系统（S。Biacchesi，M. H.斯基亚多普洛斯湾C. Tran，B. R.墨菲，P.L.柯林斯和U. J.布赫霍尔茨，Virology 321：247-259，2004）用于产生缺乏G基因、SH基因或两者的重组HMPV（rHMPV）。DeltaSH、DeltaG和DeltaSH/G缺失突变体很容易回收，并且发现在细胞培养物中的多克隆生长期间有效复制。因此，SH和G蛋白对于细胞培养物中的生长不是必需的。除了缺失的蛋白质不存在之外，基因缺失突变体产生的病毒体通过蛋白质产量和凝胶电泳蛋白质谱与野生型HMPV相似。当鼻内给予仓鼠时，DeltaG和DeltaSH/G突变体在上呼吸道和下呼吸道中复制，表明含有F作为唯一病毒表面蛋白的HMPV能够在体内复制。然而，与野生型rHMPV相比，这两种病毒在下呼吸道和上呼吸道的复制分别受到至少40倍和600倍的限制。它们还诱导高滴度的HMPV中和血清抗体，并赋予针对野生型HMPV攻击病毒在肺中复制的完全保护。令人惊讶的是，G是保护性的，DeltaG和DeltaSH/G病毒代表了有希望的疫苗候选者。相比之下，DeltaSH在仓鼠肺中的复制效率比野生型rHMPV更高（感染后第5天增加20倍）。这表明SH在体内是完全缺失的，并且其缺失不赋予减毒作用，至少在该啮齿动物模型中。
Human metapneumovirus (HMPV) has recently been identified as a significant cause of serious respiratory tract disease in humans. In particular, the emerging information on the contribution of HMPV to pediatric respiratory tract disease suggests that it will be important to develop a vaccine against this virus for use in conjunction with those being developed for human respiratory syncytial virus and the human parainfluenza viruses. A recently described reverse genetic system (S. Biacchesi, M. H. Skiadopoulos, K. C. Tran, B. R. Murphy, P. L. Collins, and U. J. Buchholz, Virology 321:247-259, 2004) was used to generate recombinant HMPVs (rHMPVs) that lack the G gene, the SH gene, or both. The DeltaSH, DeltaG, and DeltaSH/G deletion mutants were readily recovered and were found to replicate efficiently during multicycle growth in cell culture. Thus, the SH and G proteins are not essential for growth in cell culture. Apart from the absence of the deleted protein(s), the virions produced by the gene deletion mutants were similar by protein yield and gel electrophoresis protein profile to wild-type HMPV. When administered intranasally to hamsters, the DeltaG and DeltaSH/G mutants replicated in both the upper and lower respiratory tracts, showing that HMPV containing F as the sole viral surface protein is competent for replication in vivo. However, both viruses were at least 40-fold and 600-fold restricted in replication in the lower and upper respiratory tract, respectively, compared to wild-type rHMPV. They also induced high titers of HMPV-neutralizing serum antibodies and conferred complete protection against replication of wild-type HMPV challenge virus in the lungs. Surprisingly, G is dispensable for protection, and the DeltaG and DeltaSH/G viruses represent promising vaccine candidates. In contrast, DeltaSH replicated somewhat more efficiently in hamster lungs compared to wild-type rHMPV (20-fold increase on day 5 postinfection). This indicates that SH is completely dispensable in vivo and that its deletion does not confer an attenuating effect, at least in this rodent model.