Analysis of the GM-CSF and GM-CSF/IL-3/IL-5 receptor common beta chain in a patient with pulmonary alveolar proteinosis  

GM-CSF及其β_c受体在一例肺泡蛋白沉积症病人中的表达研究(英文)

作　　者：王选锭[1] 刘富光[1] Burkhard Bewig 

机构地区：[1]浙江大学医学院附属二院呼吸科,杭州310009 [2]德国基尔大学附属医院内科

出　　处：《Chinese Medical Journal》2002年第1期76-80,149,共5页中华医学杂志（英文版）

摘　　要：Objective To investigate the expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and GM-CSF/IL-3/IL-5 receptor common beta chain (βc receptor) in an adult patient with idiopathic pulmonary alveolar proteinosis (PAP), so as to demonstrate the possible association of the GM-CSF and βc receptor with the pathogenesis of human PAP.Methods The GM-CSF levels were measured with a commercial ELISA kit (sensitivity 5?pg/ml) and the βc receptor expression on the cell surface was detected by flow cytometry analysis. Reverse transcription-polymerase chain reaction (RT-PCR) analysis was employed to detect the expression of the GM-CSF mRNA and the βc receptor mRNA in peripheral blood mononuclear cells and alveolar macrophages. The entire coding regions of the GM-CSF cDNA and the βc receptor cDNA were sequenced by the Sanger dideoxy-mediated chain termination method to detect possible mutations.Results The patient with PAP failed to release the GM-CSF protein either from circulating mononuclear cells or from alveolar macrophages. The expression of the GM-CSF mRNA was normal after the stimulation of lipopolysaccharide, whereas a point mutation at position 382 of the GM-CSF cDNA from 'T' to 'C' was revealed by cDNA sequencing, which caused a change in amino acid 117 of the protein from isoleucine to threonine. The βc receptor expression on the cell surface was normal, and the βc receptor mRNA expression and the sequence of the entire coding region of the βc receptor were also normal.Conclusions The decreased GM-CSF production is associated with the pathogenesis of human PAP. A point mutation of the GM-CSF cDNA may contribute to the decreased GM-CSF production in our adult PAP patient. The mutation of the βc receptor in some of paediatric patients with PAP may not be a common problem in adult patients.目的　研究粒 巨噬细胞集落刺激因子 (GM CSF)及其βc受体在一例特发性肺泡蛋白沉积症 (PAP)成年患者中的表达 ,探索GM CSF及其βc受体与PAP发病的可能关系。方法　GM CSF蛋白测定采用ELISA法 ,白细胞表面 βc受体表达测定采用流速细胞仪。外周血单个核细胞(PBMCs)及肺泡巨噬细胞中GM CSFmRNA和 βc受体mRNA表达水平检测采用RT PCR。GM CSFcDNA和 βc受体cDNA全编码区测序采用双脱氧链终止反应法。结果　PAP患者PBMCs和肺泡巨噬细胞中均检测不到GM CSF释放 ,但GM CSFmRNA表达水平正常。cDNA测序发现PAP患者GM CSFcDNA第 382位碱基发生点突变 (TC) ,致 117位氨基酸序列发生改变 (异亮氨酸苏氨酸 )。βc受体表达、βc受体mRNA表达及其cDNA全长编码区碱基序列均无异常。结论　GM CSF蛋白表达异常与PAP发病可能有关 ,GM CSFcDNA点突变可能是导致PAP患者GM CSF蛋白表达异常的原因之一。曾在PAP患儿中发现的βc受体基因突变在本例成年PAP患者中不存在。

关 键 词：pulmonary alveolar proteinosis  ·  granulocyte-macrophage colony-stimulating factor (GM-CSF)   ·    GM-CSF/IL-3/IL-5 receptor common beta chain   ·  mutation 

分 类 号：R563.9[医药卫生—呼吸系统]