Production of Sm37-GAPDH, a major therapeutical target in human schistosomiasis.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a key enzyme in the glycolytic metabolism and the production of energy. This probably explains why GAPDH was evidenced as a major therapeutical target in several parasitic diseases; either as a vaccine candidate or as a target for chemotherapeutic treatments. Schistosoma mansoni GAPDH (Sm37-GAPDH) is one of the main schistosome vaccine candidates. The production of recombinant Sm37-GAPDH is essential to evaluate the ability of this molecule to induce protective immunity 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 production yield and the GAPDH enzymatic activity. Some of them have led to either a high production of insoluble material (E. coli) or to an inactive enzyme (Pischia pastoris). The present article describes the production setting 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 molecule and inhibited its catalytic activity.
Argiro
L
L
Laboratory of Immunology and Genetics of Parasitic Diseases, INSERM U399, Faculté de Médecine, 27 Blvd. Jean Moulin, 13385 Marseille Cedex 5, France.
Doerig
C
C
Liabeuf
S
S
Bourgois
A
A
Romette
J L
JL
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002478:Cells, Cultured; D002850:Chromatography, Gel; D004274:DNA, Recombinant / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007106:Immune Sera; D007915:Lepidoptera / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000276:immunology; D012552:Schistosomiasis / Q000276:immunology / Q000517:prevention & control*; D014614:Vaccines, Synthetic / Q000096:biosynthesis
D004274:DNA, Recombinant; D007106:Immune Sera; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases