ملخص: Human tracheal gland serous (HTGS) cells are now believed to be a major target of cystic fibrosis (CF) gene therapy. To evaluate the efficiency of adenovirus-mediated gene transfer in these cells we tested the adenovirus construction containing [beta]-galactosidase cDNA. We observed that the endogenous [beta]-galactosidase activity in cultured CF-HTGS cells was too strong to allow us to detect any exogenous [beta]-galactosidase activity. Immunohistological study on sections of human tracheal tissue confirmed the presence of [beta]-galactosidase in the serous component of the submucosal glands. We then looked for other lysosomal activities in normal and CF-HTGS cells. We showed that normal cells already have elevated enzyme values and that CF-HTGS cells contained 2-4-fold more [beta]-galactosidase, [alpha]-fucosidase, [alpha]-mannosidase and [beta]-glucuronidase activities than normal cells. An analysis of their kinetic constants has shown that this difference could be attributed to a lower Km of CF lysosomal enzymes. More importantly, these differences are eliminated after adenovirus-mediated CFTR gene transfer and not after [beta]-galactosidase gene transfer.

لغة: إنجليزية