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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。
Objective To investigate the expression of granulocyte-macrophage colony-stimulating factor (GM CSF) and its βc receptor in an adult patient with idiopathic pulmonary alveolar proteinosis (PAP) and to explore the possible relationship between GM CSF and its βc receptor and the pathogenesis of PAP . Methods GM CSF protein was determined by ELISA. The expression of βc receptor on leukocytes was measured by flow cytometry. The expression of GM CSF mRNA and βc receptor mRNA in peripheral blood mononuclear cells (PBMCs) and alveolar macrophages were detected by RT-PCR. The full coding region of GM CSF cDNA and βc receptor was sequenced by dideoxy chain termination. Results GM CSF release was not detected in PBMCs and alveolar macrophages of PAP patients, but GM CSF mRNA expression was normal. In cDNA sequencing, point mutation (TC) at base 382 of GM CSF cDNA was found in PAP patients. The 117th amino acid sequence was changed (isoleucine Threonine). βc receptor expression, βc receptor mRNA expression and its full-length cDNA coding region of the base sequence were normal. Conclusion The abnormal GM CSF protein expression may be related to the pathogenesis of PAP. Point mutation of GM CSF cDNA may be one of the reasons leading to abnormal GM CSF protein expression in PAP patients. The mutation of the βc receptor gene, found in PAP children, is absent in this case of adult PAP.