Production of Sm37-GAPDH, a major therapeutical target in human schistosomiasis

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the gly colytic metabolism and the production of energy. This probably explains why GAPDH was evidenced as a major therapeutical target in several parasitic d iseases; either as a vaccine candidate or as a target for chemotherapeutic treatments. Schistosoma mansoni GAPDH (Sm37-GAPDH) is one of the main schis tosome vaccine candidates. The production of recombinant Sm37-GAPDH is esse ntial to evaluate the ability of this molecule to induce protective immunit y in animals and possibly in humans. The cDNA encoding Sm37-GAPDH has been cloned and sequenced. In addition, five B cell (including the major B-cell epitope Sm35-5) and two T cell epitopes have been localized on the molecule . Different expression systems have been evaluated in respect with the prod uction yield and the GAPDH enzymatic activity. Some of them have led to eit her a high production of insoluble material (E. coil) or to an inactive enz yme (Pischia pastoris). The present article describes the production settin g of rSm37-GAPDH using the baculovirus-insect cell system. Large amounts of soluble rSm37-GAPDH with enzymatic activity were obtained. Most sera from individuals living in an area endemic for S. mansoni recognised the rSm37 m olecule and inhibited its catalytic activity. (C) 2000 John Wiley & Sons, I nc.