Human respiratory syncytial virus (HRSV) is an important respiratory pathogen causing a spectrum of illness, from common cold-like symptoms, to bronchiolitis and pneumonia requiring hospitalization in infants, the immunocompromised and the elderly. HRSV exists as two antigenic subtypes, A and B, which typically cycle biannually in separate seasons. There are many unresolved questions in HRSV biology regarding the interactions and interplay of the two subtypes. Therefore, we generated a reverse genetics system for a subtype A HRSV from the 2011 season (A11) to complement our existing subtype B reverse genetics system. We obtained the sequence (HRSVA11) directly from an unpassaged clinical sample and generated the recombinant (r) HRSVA11. A version of the virus expressing enhanced green fluorescent protein (EGFP) from an additional transcription unit in the fifth (5) position of the genome, rHRSVA11EGFP(5), was also generated. rHRSVA11 and rHRSVA11EGFP(5) grew comparably in cell culture. To facilitate animal co-infection studies, we derivatized our subtype B clinical isolate using reverse genetics toexpress the red fluorescent protein (dTom)-expressing rHRSVB05dTom(5). These viruses were then used to study simultaneous in vivo co-infection of the respiratory tract. Following intranasal infection, both rHRSVA11EGFP(5) and rHRSVB05dTom(5) infected cotton rats targeting the same cell populations and demonstrating that co-infection occurs in vivo. The implications of this finding on viral evolution are important since it shows that inter-subtype cooperativity and/or competition is feasible in vivo during the natural course of the infection.

中文翻译：

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重组A亚型和B亚型人呼吸道合胞病毒临床分离株共同感染棉鼠呼吸道。


人类呼吸道合胞病毒 (HRSV) 是一种重要的呼吸道病原体，可引起一系列疾病，从普通感冒样症状到需要住院治疗的婴儿、免疫功能低下者和老年人的细支气管炎和肺炎。 HRSV 以两种抗原亚型 A 和 B 的形式存在，通常在不同的季节每半年循环一次。 HRSV 生物学中关于这两种亚型之间的相互作用和相互作用还有许多未解决的问题。因此，我们为 2011 年季节 (A11) 的 A 亚型 HRSV 构建了一个反向遗传学系统，以补充我们现有的 B 亚型反向遗传学系统。我们直接从未传代的临床样本中获得了序列 (HRSV A11 )，并生成了重组 (r) HRSV A11 。还生成了表达增强型绿色荧光蛋白 (EGFP) 的病毒版本，该病毒来自基因组第五 (5) 个位置的附加转录单元 rHRSV A11 EGFP(5)。 rHRSV A11和 rHRSV A11 EGFP(5) 在细胞培养物中的生长情况相当。为了促进动物共感染研究，我们使用反向遗传学衍生化了 B 亚型临床分离株，以表达表达红色荧光蛋白 (dTom) 的 rHRSV B05 dTom(5)。然后使用这些病毒来研究呼吸道的同时体内共感染。鼻内感染后，rHRSV A11 EGFP(5) 和 rHRSV B05 dTom(5) 都感染了针对相同细胞群的棉鼠，并证明了体内发生了共感染。 这一发现对病毒进化的影响很重要，因为它表明在感染的自然过程中亚型间的合作和/或竞争在体内是可行的。