Responses of well-differentiated airway epithelial cell cultures from healthy donors and patients with cystic fibrosis to Burkholderia cenocepacia infection
Responses of well-differentiated airway epithelial cell cultures from healthy donors and patients with cystic fibrosis to Burkholderia cenocepacia infection
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DOI：
10.1128/iai.72.7.4188-4199.2004
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发表时间：
2004-07-01
影响因子：
3.1
通讯作者：
Forstner, J
中科院分区：
文献类型：
--
作者：
Sajjan, U;Keshavjee, S;Forstner, J
从囊性纤维化患者的呼吸道上皮细胞建立的分化良好的培养物(CF培养物)显示出杯状细胞增殖，粘液分泌增加，以及比来自健康捐赠者的类似培养细胞更高的白介素8基础水平。在低剂量(10(4)~10(5)cfu)的伯克霍尔德氏菌Bc7株感染根尖时，两种培养物产生不同的反应。虽然正常培养将添加的细菌捕获在粘液层，杀死和/或抑制细菌复制，并防止细菌入侵细胞，但CF培养未能杀死和/或支持细菌的生长，导致潜在的上皮细胞入侵，跨上皮通透性受损，以及细胞损伤。细菌感染前表面粘液层的耗尽使正常培养物容易受到细菌的入侵，但入侵的细菌主要局限于细胞内的空泡，似乎是不能存活的。相反，在CF培养中，侵入细胞的细菌被发现游离在中间丝包围的细胞质中，也存在于细胞之间。因此，培养的CF型呼吸道上皮比正常上皮更易感染。这模拟了活体中的CF组织，并说明了CF患者和正常受试者在处理细菌感染的方式上的差异。此外，我们还发现，Cf和正常细胞培养物不仅对分离的Bc7有不同的反应，而且对其他洋葱杆菌复合种的分离也有不同的反应。因此，我们认为该细胞培养模型适用于洋葱芽孢杆菌复合体致病机制的研究。
Well-differentiated cultures established from airway epithelia of patients with cystic fibrosis (CF cultures) exhibited goblet cell hyperplasia, increased secretion of mucus, and higher basal levels of interleukin-8 than similarly cultured cells from healthy donors. Upon apical infection with low doses (10(4) to 10(5) CFU) of Burkholderia cenocepacia isolate BC7, the two cultures gave different responses. While normal cultures trapped the added bacteria in the mucus layer, killed and/or inhibited bacterial replication, and prevented bacterial invasion of the cells, CF cultures failed to kill and/or supported the growth of bacteria, leading to invasion of underlying epithelial cells, compromised transepithelial permeability, and cell damage. Depletion of the surface mucus layer prior to bacterial infection rendered the normal cultures susceptible to bacterial invasion, but the invading bacteria were mainly confined to vacuoles within the cells and appeared to be nonviable. In contrast, bacteria that invaded cells in CF cultures were found free in the cytoplasm surrounded by intermediate filaments and also between cells. Cultured CF airway epithelium was therefore more susceptible to infection than normal epithelium. This mimics CF tissue in vivo and illustrates differences in the way epithelia in CF patients and normal subjects handle bacterial infection. In addition, we found that the CF and normal cell cultures responded differently not only to isolate BC7 but also to isolates of other B. cepacia complex species. We therefore conclude that this cell culture model is suitable for investigation of B. cepacia complex pathogenesis in CF patients.