Attenuated Human Parainfluenza Virus Type 1 (HPIV1) Expressing the Fusion Glycoprotein of Human Respiratory Syncytial Virus (RSV) as a Bivalent HPIV1/RSV Vaccine.
Attenuated Human Parainfluenza Virus Type 1 (HPIV1) Expressing the Fusion Glycoprotein of Human Respiratory Syncytial Virus (RSV) as a Bivalent HPIV1/RSV Vaccine.
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表达人呼吸道合胞病毒 (RSV) 融合糖蛋白的减毒人副流感病毒 1 型 (HPIV1)，作为二价 HPIV1/RSV 疫苗。
DOI：
10.1128/jvi.01380-15
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发表时间：
2015
影响因子：
5.4
通讯作者：
Munir,Shirin
中科院分区：
文献类型：
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作者：
Mackow,Natalie;Amaro-Carambot,Emérito;Liang,Bo;Surman,Sonja;Lingemann,Matthias;Yang,Lijuan;Collins,PeterL;Munir,Shirin
研究了重组人副流感病毒1型减毒活病毒（rHPIV 1）作为表达呼吸道合胞病毒（RSV）融合（F）糖蛋白的载体，以提供抗RSV和HPIV 1的双价疫苗。RSV F基因经工程改造以包含HPIV 1转录信号，并分别插入两个减毒rHPIV 1主链中的三个基因位置。每个骨架含有单个先前描述的减毒突变，其针对减毒稳定，具体地，涉及重叠P/C开放阅读框（ORF）中的6个核苷酸的非温度敏感性缺失突变（CΔ170）或L ORF中的温度敏感性错义突变（LY 942 A）。基因组中的插入位点是pre-N（F1）、N-P（F2）或P-M（F3），并且两个主链是相同的。在体外，F插入片段的存在降低了病毒复制速率，但最终滴度与野生型（wt）HPIV 1的最终滴度相同。在rHPIV 1-CΔ170-F1、-F2和-F3以及rHPIV 1-LY 942 A-F1中观察到培养细胞中RSV F的高水平表达。在仓鼠中，rHPIV 1-CΔ170-F1、-F2和-F3载体在鼻甲中中度受限，在肺中高度受限，并且在体内遗传稳定。在CΔ 170载体中，F1病毒对野生型RSV攻击的免疫原性和保护性最强。rHPIV 1-LY 942载体在体内是高度限制性的，并且没有可检测的免疫原性或保护性，指示过度减毒。CΔ170-F1构建体似乎是合适的减毒的和免疫原性的，以进一步开发为二价鼻内儿科疫苗。重要提示没有针对儿科呼吸道病原体RSV和HPIV的疫苗。我们正在开发用于病毒初治婴儿的RSV和HPIV减毒活疫苗。特别是减毒活RSV毒株由于其生长不良和物理不稳定性而难以开发，但这些障碍可以通过使用疫苗载体来避免。我们描述了表达RSV F蛋白的减毒rHPIV 1活载体的开发和临床前评价。将两种不同的减毒rHPIV 1主链各自工程化以从三个不同的基因位置表达RSV F。rHPIV 1-CΔ170-F1载体在P/C基因中携带减毒缺失突变（CΔ170），并从前N位表达RSV F，在仓鼠模型中对RSV F蛋白和HPIV 1具有减毒、稳定和免疫原性，并对RSV攻毒提供实质性保护。本研究提供了一种候选rHPIV 1-RSV-F疫苗病毒，适合作为针对两种主要儿童病原体的二价疫苗继续开发。
Live attenuated recombinant human parainfluenza virus type 1 (rHPIV1) was investigated as a vector to express the respiratory syncytial virus (RSV) fusion (F) glycoprotein, to provide a bivalent vaccine against RSV and HPIV1. The RSV F gene was engineered to include HPIV1 transcription signals and inserted individually into three gene locations in each of the two attenuated rHPIV1 backbones. Each backbone contained a single previously described attenuating mutation that was stabilized against deattenuation, specifically, a non-temperature-sensitive deletion mutation involving 6 nucleotides in the overlapping P/C open reading frames (ORFs) (CΔ170) or a temperature-sensitive missense mutation in the L ORF (LY942A). The insertion sites in the genome were pre-N (F1), N-P (F2), or P-M (F3) and were identical for both backbones.In vitro, the presence of the F insert reduced the rate of virus replication, but the final titers were the same as the final titer of wild-type (wt) HPIV1. High levels of RSV F expression in cultured cells were observed with rHPIV1-CΔ170-F1, -F2, and -F3 and rHPIV1-LY942A-F1. In hamsters, the rHPIV1-CΔ170-F1, -F2, and -F3 vectors were moderately restricted in the nasal turbinates, highly restricted in lungs, and genetically stablein vivo. Among the CΔ170vectors, the F1 virus was the most immunogenic and protective against wt RSV challenge. The rHPIV1-LY942Avectors were highly restrictedin vivoand were not detectably immunogenic or protective, indicative of overattenuation. The CΔ170-F1 construct appears to be suitably attenuated and immunogenic for further development as a bivalent intranasal pediatric vaccine.IMPORTANCEThere are no vaccines for the pediatric respiratory pathogens RSV and HPIV. We are developing live attenuated RSV and HPIV vaccines for use in virus-naive infants. Live attenuated RSV strains in particular are difficult to develop due to their poor growth and physical instability, but these obstacles could be avoided by the use of a vaccine vector. We describe the development and preclinical evaluation of live attenuated rHPIV1 vectors expressing the RSV F protein. Two different attenuated rHPIV1 backbones were each engineered to express RSV F from three different gene positions. The rHPIV1-CΔ170-F1 vector, bearing an attenuating deletion mutation (CΔ170) in the P/C gene and expressing RSV F from the pre-N position, was attenuated, stable, and immunogenic against the RSV F protein and HPIV1 in the hamster model and provided substantial protection against RSV challenge. This study provides a candidate rHPIV1-RSV-F vaccine virus suitable for continued development as a bivalent vaccine against two major childhood pathogens.