High lysosomal activities in cystic fibrosis tracheal gland cells corrected by adenovirus-mediated CFTR gene transfer.
1999
Abstract 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 β-galactosidase cDNA. We observed that the endogenous β-galactosidase activity in cultured CF-HTGS cells was too strong to allow us to detect any exogenous β-galactosidase activity. Immunohistological study on sections of human tracheal tissue confirmed the presence of β-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 β-galactosidase, α-fucosidase, α-mannosidase and β-glucuronidase activities than normal cells. An analysis of their kinetic constants has shown that this difference could be attributed to a lower K m of CF lysosomal enzymes. More importantly, these differences are eliminated after adenovirus-mediated CFTR gene transfer and not after β-galactosidase gene transfer.
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