Over-expression of neuropathy target esterase activity in bovine chromaffin cell cultures by adenovirus-mediated gene transfer

Abstract: Chromaffin cells in culture show high neuropathy target esterase (NTE) activity. It is well known that inhibition and specific modification of NTE by some organophosphorus (OPs) compounds induces a neurodegenerative neuropathy. It has been suggested that NTE is responsible for phosphatidylcholine homeostasis, although its role in neuropathy induction remains unclear. The cDNA of human NTE (4.4kbp) was inserted into an adenoviral vector. Bovine chromaffin cells cultured at 50,000 cells/well were incubated with the vector for 2h and after removing the volume of infection, cells were maintained in the incubator. After 24h, NTE activity was 6.8±0.5mU/106 cells in untreated cells and 14.8±1.5mU/106 cells, 19.3±2.9mU/106 cells, 24.8±0.9mU/106 cells and 30.9±1.0mU/106 cells in cells incubated with 2, 4, 8 and 16μl of vector, respectively. After 60min of inhibition with mipafox increased concentrations, the calculated I50 (60min) values were 5.5, 6.2 and 6.6μM for cells infected with 0, 2 and 10μl of vector preparation. We confirm that the adenoviral vector containing the human NTE gene is active in bovine chromaffin cells in culture and that the NTE activity expressed by the vector shows the same inhibition pattern by the neuropathic OP mipafox as the NTE activity of bovine chromaffin cells and cells remained viable after the high NTE activity expression. [Copyright &y& Elsevier]