Abstract：
Respiratory syncytial virus (RSV) remains the primary cause of lower respiratory tract infections, particularly in infants and the elderly. In this study, we employed reverse genetics to generate a chimeric influenza virus expressing neuraminidase-3F protein conjugate with three repeats of the RSV F protein protective epitope inserted into the NA gene of A/California/7/2009 ca (CA/AA ca), resulting in rFlu/RSV/NA-3F (hereafter, rFRN3). The expression of NA-3F protein was confirmed by Western blotting. The morphology and temperature-sensitive phenotype of rFRN3 were similar to CA/AA ca. Its immunogenicity and protective efficiency were evaluated in BALB/c mice and cotton rats. Intranasal administration of rFRN3 elicited robust humoral, cellular, and to some extent, mucosal immune responses. Compared to controls, rFRN3 protected animals from RSV infection, attenuated lung injury, and reduced viral titers in the nose and lungs post-RSV challenge. These results demonstrate that rFRN3 can trigger RSV-specific immune responses and thus exhibits potent protective efficacy. The \"dual vaccine\" approach of a cold-adapted influenza vector RSV vaccine will improve the prophylaxis of influenza and RSV infection. rFRN3 thus warrants further clinical investigations as a candidate RSV vaccine.
摘要：
呼吸道合胞病毒（RSV）仍然是下呼吸道感染的主要原因，尤其是婴儿和老人。在这项研究中,我们使用反向遗传学来产生表达神经氨酸酶-3F蛋白缀合物的嵌合流感病毒，该缀合物具有插入A/California/7/2009ca（CA/AAca）的NA基因中的RSVF蛋白保护性表位的三个重复序列，导致rFlu/RSV/NA-3F(以下，rFRN3).通过蛋白质印迹证实NA-3F蛋白的表达。rFRN3的形态和温度敏感表型与CA/AAca相似。在BALB/c小鼠和棉鼠中评价其免疫原性和保护效率。rFRN3的鼻内给药引起强健的体液，细胞，在某种程度上,粘膜免疫反应。与对照组相比，rFRN3保护动物免受RSV感染，减轻肺损伤，RSV攻击后鼻和肺中的病毒滴度降低。这些结果证明rFRN3可以触发RSV特异性免疫应答,因此表现出有效的保护功效。冷适应流感载体RSV疫苗的“双重疫苗”方法将改善流感和RSV感染的预防。因此,rFRN3作为候选RSV疫苗需要进一步的临床研究。
