Glycobiology, 2002, Vol. 12, No. 2 135-144
© 2002 Oxford University Press
Expression of 
-gal epitopes on HeLa cells transduced with adenovirus containing 1,3galactosyltransferase cDNA
2Departments of Cardiovascular-Thoracic Surgery and Immunology, Rush University, 1653 West Congress Parkway, Chicago, IL 60612, USA and 3Institute of Gene Therapy, University of Pennsylvania, M6. 30 Malony, Philadelphia, PA 19104, USA
1,3Galactosyltransferase (1,3GT) synthesizes -gal epitopes (Gal1-3Galß1-4GlcNAc-R) on glycoconjugates in nonprimate mammals but not in humans. Transduction of 1,3GT gene into human HeLa cells by an adenovirus vector allowed for accurate kinetics studies on the appearance of 1,3GT and of its product, the -gal epitope, in the transduced cells. Mouse 1,3GT cDNA was inserted into a replication-defective adenovirus vector. This viral vector, designated AdGT, could be propagated in human 293 cells that have the viral E1 complementing gene. Transduction of HeLa cells resulted in immediate penetration of 
20 AdGT copies into each cell and the appearance of 1,3GT mRNA after 4h. Catalytic activity of 1,3GT was first detected in the cells after 6 h. The initial appearance of -gal epitopes (6 x 104/cell) on cell surface glycoconjugates was detected 10 h posttransduction, whereas 24 h posttransduction each cell expressed 2 x 106 epitopes. The activity of 1,3GT in cells transduced with approximately two copies of AdGT was eightfold lower than that in cells transduced with 20 AdGT copies; however, the number of -gal epitopes/cell remained closely similar. This implies that increased 1,3GT activity above a certain saturation level does not result in a corresponding increase in the carbohydrate product, possibly because of competing glycosyltransferases.
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