Forrest I A, Murphy D M, Ward C, Jones D, Johnson G E, Archer L, Gould F K, Cawston T E, Lordan J L, Corris P A
Applied Immunobiology and Transplantation Research Group, University of Newcastle upon Tyne, UK.
Eur Respir J. 2005 Dec;26(6):1080-5. doi: 10.1183/09031936.05.00141404.
Long-term survival in lung transplantation is limited by the development of obliterative bronchiolitis, a condition characterised by inflammation, epithelial injury, fibroproliferation and obliteration of bronchioles leading to airflow obstruction. To investigate the role of the bronchial epithelium in the pathogenesis of obliterative bronchiolitis the current study aimed to establish primary bronchial epithelial cell cultures (PBEC) from lung allografts. Four to six bronchial brushings were obtained from sub-segmental bronchi of lung allografts. Cells were seeded onto collagen-coated plates and grown to confluence in bronchial epithelial growth medium. Bronchial brushings (n=33) were obtained from 27 patients. PBECs were grown to confluence from 12 out of 33 (39%) brushings. Failure to reach confluence was due to early innate infection. Bacteria were usually isolated from both bronchoalveolar lavage and culture media, but a separate population was identified in culture media only. Primary culture of bronchial epithelial cells from lung transplant recipients is feasible, despite a high rate of early, patient-derived infection. Latent infection of the allograft, identified only by bronchial brushings, may itself be a persistent stimulus for epithelial injury. This technique facilitates future mechanistic studies of airway epithelial responses in the pathogenesis of obliterative bronchiolitis.
肺移植的长期存活受到闭塞性细支气管炎发展的限制,这种病症的特征是炎症、上皮损伤、纤维增殖以及细支气管闭塞导致气流阻塞。为了研究支气管上皮在闭塞性细支气管炎发病机制中的作用,本研究旨在从肺移植供体建立原代支气管上皮细胞培养物(PBEC)。从肺移植供体的亚段支气管获取4至6次支气管刷检样本。将细胞接种到胶原包被的培养板上,并在支气管上皮生长培养基中培养至汇合。从27例患者中获取了33次支气管刷检样本。33次刷检样本中有12次(39%)的PBEC培养至汇合。未能达到汇合是由于早期先天性感染。细菌通常从支气管肺泡灌洗样本和培养基中分离出来,但仅在培养基中鉴定出一个单独的菌群。尽管早期患者源性感染率很高,但从肺移植受者中进行支气管上皮细胞原代培养是可行的。仅通过支气管刷检鉴定出的移植肺潜在感染本身可能是上皮损伤的持续刺激因素。这项技术有助于未来对闭塞性细支气管炎发病机制中气道上皮反应进行机制研究。