Tristram D A, Hicks W, Hard R
Department of Pediatrics, State University of New York at Buffalo School of Medicine, Children's Hospital of Buffalo, 14222, USA.
Arch Otolaryngol Head Neck Surg. 1998 Jul;124(7):777-83. doi: 10.1001/archotol.124.7.777.
A suitable model for respiratory syncytial virus (RSV) infection has yet to be developed.
To describe an in vitro model of human respiratory epithelium in primary cell culture linked with a computer microscope interface that allows evaluation and imaging of living RSV-infected respiratory epithelium.
A descriptive, controlled study. Human bronchial cells were obtained from surgical samples by elastase dissociation and replated on collagen gel membranes. After 7 to 10 days, cells were brought to air interface. Baseline sampling of cell fluid for cytokine production by enzyme-linked immunosorbent assay (interleukin [IL] 1beta, IL-6, IL-8, and RANTES) and leukotriene C4 by radioimmunoassay was taken before treatment with RSV (n=30) or HEp-2 (human laryngeal carcinoma cells) control (n=25). Sampling was done at 4, 24, 72, and 120 hours thereafter. The infectious process was monitored with a microscope (Zeiss UEM, Carl Zeiss Inc, Thornwood, NY) equipped with a camera (Newvicon, Dage Corporation, Stamford, Conn). Images were either digitized using a computer (Macintosh Quadra 950, Apple Computers Inc, Cupertino, Calif) equipped with a digitizing board (Perceptics Corporation, Knoxville, Tenn) or were recorded on an SVHS videotape using a videocassette recorder (JVC, Elmwood Park, NJ).
Respiratory syncytial virus induced profound effects on the ciliated cells: ciliostasis, clumping, and loss of cilia from live cells and sloughing of cells. Significant differences in the release of IL-6, IL-8, and RANTES (P<.03 for each cytokine) were noted in RSV-infected bronchial cultures by 24 hours with a peak at 72 hours. The IL-beta and leukotriene C4 were not altered by RSV infection in bronchial cells.
This model closely mirrors human RSV disease and affords a unique opportunity to study interepithelial cell interactions, cytokine responses from cells of different donors, and ciliary activity of live cells undergoing RSV infection.
呼吸道合胞病毒（RSV）感染的合适模型尚未建立。
描述一种原代细胞培养中的人呼吸道上皮体外模型，该模型与计算机显微镜接口相连，可对感染RSV的活呼吸道上皮进行评估和成像。
一项描述性对照研究。通过弹性蛋白酶解离从手术样本中获取人支气管细胞，并重新接种于胶原凝胶膜上。7至10天后，使细胞处于气液界面。在用RSV（n = 30）或HEp - 2（人喉癌细胞）对照（n = 25）处理前，通过酶联免疫吸附测定法（白细胞介素[IL] - 1β、IL - 6、IL - 8和调节激活正常T细胞表达和分泌因子[RANTES]）对细胞培养液进行细胞因子产生的基线采样，并用放射免疫测定法对白三烯C4进行采样。此后在4、24、72和120小时进行采样。使用配备相机（Newvicon，Dage公司，斯坦福德，康涅狄格州）的显微镜（蔡司UEM，卡尔蔡司公司，索恩伍德，纽约州）监测感染过程。图像要么使用配备数字化板（Perceptics公司，诺克斯维尔，田纳西州）的计算机（苹果麦金塔Quadra 950，苹果电脑公司，库比蒂诺，加利福尼亚州）进行数字化，要么使用盒式录像机（JVC，埃尔姆伍德公园，新泽西州）记录在超级 VHS录像带上。
呼吸道合胞病毒对纤毛细胞产生深远影响：纤毛静止、聚集、活细胞纤毛丧失以及细胞脱落。在RSV感染的支气管培养物中，到24小时时，IL - 6、IL - 8和RANTES的释放出现显著差异（每种细胞因子P <.03），在72小时达到峰值。支气管细胞中的IL - β和白三烯C4未因RSV感染而改变。
该模型紧密模拟人RSV疾病，为研究上皮细胞间相互作用、不同供体细胞的细胞因子反应以及感染RSV的活细胞的纤毛活动提供了独特机会。
