ANALYSIS OF MACROPHAGE SPECIFICalpha1-ANTITRYPSIN PROMOTER IN PATIENTS WITH EMPHYSEMA

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The purpose of this study is to investigate the genetic factors for pulmonary emphysema. We analyzed the genetic polymorphism of macrophage specific promoter sequences of alpha1-antitrypsin gene. Mononuclear cells were separated from peripheral blood of patients with pulmonary emphysema, chronic bronchiolitis and bronchial asthma, and the genomic DNA was isolated. Macrophage specific promoter sequences of alpha1-antitrypsin gene was amplified by polymerase chain reaction. Single strand conformational polymorphism analysis was adopted on the analysis of the amplified DNA sequences. We detected three types of genetic polymorphism of macrophage specific promoter sequences of alpha1-antitrypsin gene by single-strand conformational polymorphism analysis. The genetic polymorphism was not associated with the diseases which were investigated. Then, we cloned the cDNA of macrophage specific promoter sequences of alpha1-antitrypsin gene from the normal subjects. The sequences were inserted into pGL2-basic vector, which contains luciferase gene as a reporter gene to test the unknown promoter activity. Our results demonstrated that macrophage specific promoter sequences of alpha1-antitrypsin gene showed the weak promoter activity itself, and the macrophage specific promoter activity was not clear. These findings suggest that further investigation would be necessary to study the high activity and macrophage specificity of the promoter sequences for alpha1-antitrypsin gene after stimulation of the gene transfected cells by cytokines or other factors.