Preliminary characterization of an adult worm "vomit" preparation of Schistosoma mansoni and its potential use as antigen for diagnosis.
Schistosoma mansoni is a parasitic trematode of the portal-mesenteric veins with a closed-end intestine. Adult worms regurgitate their intestinal content after digestion, together with constituents of the lining gut. Some of these molecules circulate in the blood and are antigenic. We obtain a "vomit" preparation and preliminary evaluate its biochemical composition and antigenic capacity. The "vomit" preparation was obtained after changes in temperature and solutions of incubation of adult worms between 4 and 37 degrees C. Supernatant was assayed for protein, carbohydrate concentration and enzymatic activities associated to the intestine and to the worm tegument. The antigenicity of the product was evaluated using Western blot (WB) analysis against sera of experimentally infected mice, before and after drug cure, sera from people infected with S. mansoni and from individuals infected with other parasitoses. More carbohydrate than protein was detected in the preparations. Cysteine proteinase (CP), N-acetyl-beta-D: -glucosaminidase and alkaline phosphatase activities were detected. The latter enzyme activity is a marker of the tegument, suggesting that in spite of careful conditions used to avoid the presence of tegumental material, manipulation of the worms always resulted in the release of tegumental molecules. Cationic exchange chromatography was useful to separate various components of this "vomit" preparation, particularly enzymes responsible for CP activity. Two highly immunogenic and specific duplets were observed in the WB analysis, 31/32- and 38/40-kDa components, the former probably referring to the intestinal CPs Sm31/Sm32. None of the two duplets disappeared after successful chemotherapy during the time of evaluation in mice or humans.
Planchart
Sandra
S
Departamento de Parasitología, Facultad de Ciencias de la Salud, Universidad de Carabobo, Valencia, Venezuela.
Incani
Renzo Nino
RN
Cesari
Italo Mario
IM
D000118:Acetylglucosaminidase / Q000032:analysis; D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000737:chemistry / Q000302:isolation & purification*; D015153:Blotting, Western; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000032:analysis; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D012550:Schistosoma mansoni / Q000737:chemistry / Q000276:immunology*; D012555:Schistosomiasis mansoni / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D000469:Alkaline Phosphatase; D000118:Acetylglucosaminidase; D003546:Cysteine Endopeptidases