Laboratory of Experimental Immunology, Catholic University Hospitals, Leuven, Belgium.
Rhinology. 2010 Dec;48(4):401-7. doi: 10.4193/Rhino10.033.
Human nasal epithelial cells (hNECs) are the first line of immune defense and are able to produce mediators that recruit, activate and prolong survival of immune cells, among which IL-8 takes an important place. Studies on IL-8 and effects of dexamethasone on hNECs have been hampered by methodological shortcomings. The purpose of the study is to investigate the contribution of freshly isolated hNECs to IL-8 production in chronic rhinosinusitis with nasal polyps (CRSwithNP). Secondly, the effects of dexamethasone treatment on hNEC apoptosis and IL-8 production are investigated.
hNECs were freshly isolated from nasal polyp tissue and healthy inferior turbinate of NP patients (n=12) and from inferior turbinates of healthy donors (n=19) by protease treatment and two negative selection procedures. hNECs were incubated with IL-1β (10ng/ml), TNFα (10ng/ml) or dexamethasone (10, 100 and 1000 Amicrog/ml). After 24h, IL-8 levels were determined in the supernatants by ELISA. Finally, hNECs were incubated with increasing doses of dexamethasone and stained with trypan-blue and annexin-FITC/PI to study apoptosis.
hNECs isolated from nasal turbinates of healthy and NP patients and polyp tissue from NP patients produced similar levels of IL-8. IL-1β induced higher levels of IL-8 production in all types of hNECs without differences between control and NP tissue. Dexamethasone induced apoptosis of hNECs concomitant with abrogation of IL-8 production by hNECs.
IL-8 production by human nasal epithelial cells does not differ between NP and healthy tissue under baseline nor stimulatory conditions. Dexamethasone induces apoptosis of hNECs and abrogates IL-8 production.
人鼻腔上皮细胞(hNEC)是第一道免疫防线,能够产生招募、激活并延长免疫细胞存活的介质,其中 IL-8 起着重要作用。由于方法学上的缺陷,IL-8 及地塞米松对 hNEC 的作用的研究受到了阻碍。本研究旨在探讨新鲜分离的 hNEC 在伴有鼻息肉的慢性鼻-鼻窦炎(CRSwNP)中产生 IL-8 的作用。其次,研究地塞米松处理对 hNEC 凋亡和 IL-8 产生的影响。
蛋白酶处理和两次阴性选择程序从 NP 患者的鼻息肉组织和下鼻甲(n=12)以及健康供体的下鼻甲(n=19)中新鲜分离 hNEC。将 hNEC 与 IL-1β(10ng/ml)、TNFα(10ng/ml)或地塞米松(10、100 和 1000Amicrog/ml)孵育。24 小时后,通过 ELISA 测定上清液中的 IL-8 水平。最后,用递增剂量的地塞米松孵育 hNEC,并用台盼蓝和 Annexin-FITC/PI 染色以研究凋亡。
从健康和 NP 患者的鼻甲和 NP 患者的息肉组织中分离的 hNEC 产生相似水平的 IL-8。IL-1β 在所有类型的 hNEC 中诱导更高水平的 IL-8 产生,且在对照和 NP 组织之间无差异。地塞米松诱导 hNEC 凋亡,同时阻断 hNEC 的 IL-8 产生。
在基线和刺激条件下,NP 和健康组织中的人鼻腔上皮细胞产生的 IL-8 没有差异。地塞米松诱导 hNEC 凋亡并阻断 IL-8 产生。