Mosler Katharina, Coraux Christelle, Fragaki Konstantina, Zahm Jean-Marie, Bajolet Odile, Bessaci-Kabouya Katia, Puchelle Edith, Abély Michel, Mauran Pierre
INSERM, U514, Reims, F-51092, France.
J Cyst Fibros. 2008 Jan;7(1):44-53. doi: 10.1016/j.jcf.2007.04.005. Epub 2007 Jun 5.
For a better understanding of the early stages of cystic fibrosis (CF), it is of major interest to study respiratory epithelial cells obtained as early as possible. Although bronchoalveolar lavage has been proposed for this purpose, nasal brushing, which is a much less invasive technique, has seldom been used in CF infants. The aim of the present study was to examine in a few infants the feasibility of a nasal brushing technique for studies of airway epithelial functions in very young CF infants.
In 5 CF (median age 12, range 1-18 months) and 10 control infants (median age 5, range 1-17 months), a nasal brushing was performed by means of a soft sterile cytology brush, after premedication with oral paracetamol (15 mg/kg body weight) and rectal midazolam (0.2 mg/kg body weight). Samples were used for microbiological, cytological and functional studies.
The procedure was well tolerated. Number of cells collected was similar in CF and non-CF patients (CF: median 230x10(3), range 42x10(3)-900x10(3); non-CF: median 340x10(3), range 140x10(3)-900x10(3)). Median number of viable cells was 67% (range 31-84%). Freshly obtained samples were successfully used for studies of ciliary beating frequency and cAMP-dependent chloride efflux. In 7 out of 17 cell cultures, confluence was obtained (CF: 2 out of 7; non-CF: 5 out of 10). The feasibility of studying protein release and mRNA expression of IL-8, IL-6 and TNF-alpha, under basal conditions and after stimulation by Pseudomonas aeruginosa, was demonstrated.
By means of a simple nasal brushing technique easily performed and well tolerated, it is feasible, in infants, to harvest respiratory cells in sufficient amounts to study the airway epithelium using a broad range of techniques including cell culture.
为了更好地了解囊性纤维化(CF)的早期阶段,尽早获取呼吸道上皮细胞进行研究具有重要意义。虽然已有人提出使用支气管肺泡灌洗来实现这一目的,但鼻拭子采集是一种侵入性小得多的技术,在CF婴儿中很少使用。本研究的目的是在少数婴儿中检验鼻拭子采集技术用于研究极年幼CF婴儿气道上皮功能的可行性。
对5名CF婴儿(中位年龄12个月,范围1 - 18个月)和10名对照婴儿(中位年龄5个月,范围1 - 17个月),在口服对乙酰氨基酚(15毫克/千克体重)和直肠给予咪达唑仑(0.2毫克/千克体重)进行预处理后,用软质无菌细胞学刷进行鼻拭子采集。采集的样本用于微生物学、细胞学和功能研究。
该操作耐受性良好。CF患者和非CF患者采集到的细胞数量相似(CF:中位值230×10³,范围42×10³ - 900×10³;非CF:中位值340×10³,范围140×10³ - 900×10³)。活细胞的中位数量为67%(范围31% - 84%)。新采集的样本成功用于纤毛摆动频率和环磷酸腺苷(cAMP)依赖性氯离子外流的研究。在17个细胞培养物中有7个达到汇合状态(CF:7个中有2个;非CF:10个中有5个)。证明了在基础条件下以及铜绿假单胞菌刺激后研究白细胞介素 - 8(IL - 8)、白细胞介素 - 6(IL - 6)和肿瘤坏死因子 - α(TNF - α)的蛋白质释放和mRNA表达的可行性。
通过一种易于操作且耐受性良好的简单鼻拭子采集技术,在婴儿中采集足够数量的呼吸道细胞以使用包括细胞培养在内的多种技术研究气道上皮是可行的。