The Regulatory Effects of Human Bronchial Fibroblasts on Peripheral Blood Eosinophils: An In Vitro Study

人類支氣管纖維母細胞對週邊血液嗜伊紅性白血球的體外調節作用

楊宗穎(Tsung-Ying Yang) ； 許正園(Jeng-Yuan Hsu) ； 朱兆嘉(Jao-Jia Chu) ； 江自得(Chi-Der Chiang)

《胸腔醫學》 18卷3期 (2003/06) Pp. 194-198

https://doi.org/10.29806/TM.200306.0002

摘要

氣喘是一個慢性氣道炎性疾病，嗜酸性白血球在組織內的存在及活化與氣喘的疾病機制息息相關。曾有報告顯示纖維母細胞可以影響嗜酸性白血球的功能，本研究主旨即在探討這兩種細胞間的交互關係。
方法：人類支氣管纖維母細胞細胞株CCD14-Br購自ATCC並保存於含10 %胎牛血清之MEM培養液。氣喘病人之週邊血液經抽取後以f-met-leu-phe（fmlp）處理並用Percoll比重離心法製成人類周邊血液嗜酸性白血球。細胞存活率分析採用MTT比色法，流式細胞儀則用來分析經螢光染色後的細胞表面抗原表現情形。
結果：經分離後的人類周邊血液嗜酸性白血球純度可達85%。超過一半的嗜酸性白血球在體外無法存活超過三天以上。將嗜酸性白血球與纖維母細胞一齊培養至第七天時，可發現將近60 %的嗜酸性白血球仍然存活。若於最初同時加入50 pM人類重組顆粒球生成素，有80 %的嗜酸性白血球可存活七天；但是若只加入重組顆粒球生成素而無纖維母細胞，則第七天時只有50 %之纖維母細胞存活。使用Transwella半透膜阻斷纖維母細胞與嗜酸性白血球的直接接觸，於第五天時嗜酸性白血球的細胞活率由63 %降為45.8 %。以流式細胞儀觀察，嗜酸性白血球與CCD14-Br一齊培養後，HLA-DR表面抗原之表現明顯增加，但CD4表面抗原則無任何改變。
結論：本研究之結果顯示支氣管纖維母細胞可經由控制嗜酸性白血球的存活進一步影響氣喘的病態生理變化。

關鍵字

氣喘 ； 嗜酸性白血球 ； 纖維母細胞

並列摘要

Background: Asthma is a chronic inflammatory disease of the airways. Its disease status is closely associated with the survival and activation of eosinophils. Because it has been reported that fibroblasts partially govern the regulatory functions of eosinophils, we were interested in investigating the relationship between the two cells.
Methods: CCD14-Br human bronchial fibroblast cells were obtained from the American Type Culture Collection (ATCC) and maintained in a MEM medium containing 10% fetal calf serum. Human peripheral blood eosinophils were isolated from f-met-leu-phe (fmlp) treated blood and followed by discontinuous Percoll density gradient centrifugation. Cell survival was determined with MTT colorimetric analysis. The expression of cell surface markers was analyzed by employing immunofluorescence staining and quantitated with a fluorescence activated cell sorter.
Results: The purity of eosinophil isolation exceeded 85%. More than half of the isolated eosinophils did not survive three days in vitro. If we cultured the eosinophils with CCD14-Br cells, 60% survived at the seventh day, and 80% survived in the presence of 50 pM recombinant human GM-CSF. Only 50% of eosinophils survived in the treatment of GM-CSF without CCD14-Br cells. Blocking the contact between the two cells by employing Transwella, the survival of eosinophils significantly decreased as compared with that of the direct cocultured system (45.8% vs. 63%) at the fifth day. The expression of HLA-DR markedly increased, whereas that of CD4 remained unchanged when cocultured with CCD14-Br cells.
Conclusion: The results indicated that bronchial fibroblasts might play a role in the pathogenesis of asthma via regulating the life span of eosinophils.

並列關鍵字

asthma ； eosinophils ； fibroblasts