Institute of Translational Medicine (Child Health), University of Liverpool, Alder Hey Children's Hospital, Eaton Road, Liverpool L12 2AP, UK.
Thorax. 2012 Jan;67(1):42-8. doi: 10.1136/thoraxjnl-2011-200131. Epub 2011 Aug 23.
Respiratory syncytial virus (RSV) infection of airway epithelial cells (AECs) is an important initial event in RSV bronchiolitis. AEC immunological responses are thought to be critical in driving the subsequent inflammation in the airway. This study examined viral replication, cytotoxicity and cytokine production in cultures of primary AECs from children compared with responses to RSV infection in an immortalised epithelial cell line and to those from infants with RSV bronchiolitis.
RSV replication, proinflammatory cytokine responses and cytotoxicity in RSV-infected primary AEC cultures derived from bronchial brushings from the lungs of children were compared with those seen in BEAS-2B cultures, as well as AECs and bronchoalveolar lavage fluid collected from children with and without RSV bronchiolitis.
Viral replication, cytotoxicity and inflammatory cytokine production were greater in primary AEC cultures than in BEAS-2B cells. Different response patterns were observed, with RSV infection of primary AEC cultures causing distinct peaks of viral replication and matched cytotoxic responses. Some primary AEC culture immunological responses, such as interleukin 8, were similar in magnitude to those seen in clinical samples from the lungs of children with RSV bronchiolitis. Although variable amounts of RSV were detected by PCR in freshly isolated primary AECs, RSV was not detected by immunocytochemistry.
This is one of the first studies to examine comprehensively the responses to RSV infection in primary AEC cultures from children and shows marked differences from those of a commercially available immortalised human cell line but reassuring similarities to results found in vivo. This suggests that future work investigating responses of AECs to RSV infection should use primary AEC cultures.
呼吸道合胞病毒(RSV)感染气道上皮细胞(AEC)是 RSV 细支气管炎的重要初始事件。AEC 的免疫反应被认为是驱动随后气道炎症的关键因素。本研究比较了儿童原代 AEC 培养物与永生化上皮细胞系以及 RSV 细支气管炎患儿的 AEC 和支气管肺泡灌洗液对 RSV 感染的病毒复制、细胞毒性和细胞因子产生的反应,以研究其反应。
比较从儿童肺部支气管刷取物中分离的 RSV 感染原代 AEC 培养物与 BEAS-2B 培养物、以及患有和不患有 RSV 细支气管炎的儿童的 AEC 和支气管肺泡灌洗液中观察到的 RSV 复制、促炎细胞因子反应和细胞毒性。
原代 AEC 培养物中的病毒复制、细胞毒性和炎症细胞因子产生均高于 BEAS-2B 细胞。观察到不同的反应模式,RSV 感染原代 AEC 培养物导致病毒复制和匹配的细胞毒性反应出现明显峰值。一些原代 AEC 培养物免疫反应,如白细胞介素 8,其大小与 RSV 细支气管炎患儿肺部临床样本中观察到的相似。尽管通过 PCR 在新鲜分离的原代 AEC 中检测到了不同量的 RSV,但通过免疫细胞化学未检测到 RSV。
这是首次全面研究儿童原代 AEC 培养物对 RSV 感染的反应的研究之一,与商业上可用的永生化人细胞系的反应存在明显差异,但与体内研究结果相似。这表明,未来研究 AEC 对 RSV 感染的反应应该使用原代 AEC 培养物。