Bronchoalveolar lavage (BAL) for research; obtaining adequate sample yield.
Bronchoalveolar lavage (BAL) for research; obtaining adequate sample yield.
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支气管肺泡灌洗（BAL）用于研究；获得足够的样品产量。
DOI：
10.3791/4345
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发表时间：
2014-03-24
期刊：
影响因子：
--
通讯作者：
Gordon SB
中科院分区：
文献类型：
--
作者：
Collins AM;Rylance J;Wootton DG;Wright AD;Wright AK;Fullerton DG;Gordon SB
我们描述了一种使用手动手持吸力进行支气管肺泡灌洗（BAL）的纤维支气管镜检查的研究技术，以去除粘膜表面的非粘附细胞和肺内壁液体。在研究环境中，BAL 允许对肺内的先天（肺巨噬细胞）、细胞（B 细胞和 T 细胞）和体液（免疫球蛋白）反应进行采样。 BAL 已被国际认可用于研究目的，自 1999 年以来，我们的团队已在英国和马拉维对超过 1,000 名受试者进行了这项技术。我们的技术采用手持式轻轻抽吸滴注液；其目的是最大化返回的 BAL 体积并对纤毛上皮施加最小的剪切力，以保留 BAL 液内细胞的结构和功能，并保持活力以促进离体培养中的细胞生长。因此，该研究技术使用较大体积的滴注液（通常为 200 毫升左右）并采用手动抽吸来减少细胞损伤。患者接受局部麻醉，并给予清醒镇静（咪达唑仑），并且能够很好地耐受手术，副作用最小。口头和书面的受试者信息可以提高耐受性，并且书面知情同意是强制性的。受试者的安全至关重要。使用明确的纳入和排除标准仔细选择受试者。该协议包括对潜在风险的描述以及减轻风险所采取的步骤、禁忌症列表、手术前和手术后检查以及精确的支气管镜检查和实验室技术。
We describe a research technique for fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) using manual hand held suction in order to remove nonadherent cells and lung lining fluid from the mucosal surface. In research environments, BAL allows sampling of innate (lung macrophage), cellular (B- and T- cells), and humoral (immunoglobulin) responses within the lung. BAL is internationally accepted for research purposes and since 1999 the technique has been performed in > 1,000 subjects in the UK and Malawi by our group. Our technique uses gentle hand-held suction of instilled fluid; this is designed to maximize BAL volume returned and apply minimum shear force on ciliated epithelia in order to preserve the structure and function of cells within the BAL fluid and to preserve viability to facilitate the growth of cells in ex vivo culture. The research technique therefore uses a larger volume instillate (typically in the order of 200 ml) and employs manual suction to reduce cell damage. Patients are given local anesthetic, offered conscious sedation (midazolam), and tolerate the procedure well with minimal side effects. Verbal and written subject information improves tolerance and written informed consent is mandatory. Safety of the subject is paramount. Subjects are carefully selected using clear inclusion and exclusion criteria. This protocol includes a description of the potential risks, and the steps taken to mitigate them, a list of contraindications, pre- and post-procedure checks, as well as precise bronchoscopy and laboratory techniques.