Study of the in vitro activation of the complement alternative pathway by Echinococcus granulosus hydatid cyst fluid.
In the present study we have investigated the fluid phase activation of the complement (C) alternative pathway by Echinococcus granulosus sheep hydatid cyst fluid (SHCF) and its higher molecular weight fraction (SHCF-I) by quantitating the formation of both the terminal C intermediary C5b6 complex and the terminal C complex (TCC). Our results show that in vitro C activation progresses beyond the C5 step suggesting that potentially lytic complexes may be generated in vivo. In addition, SHCF and SHCF-I glucidic moieties are probably involved in C activation since 80% and 86% of SHCF and SHCF-I activity respectively was destroyed by periodate oxidation. Furthermore, partial deglycosylation with Peptide N-Glycosidase F of SHCF-I which had been digested with Pronase E, released an active fraction (MW < 14 KDa) which bound to Soybean agglutinin, suggesting that N-linked oligosaccharides containing alpha- or beta-linked N-acetyl galactosamine play a role in C activation by SHCF.
Ferreira
A M
AM
Molecular Immunopathology Unit, MRC Centre, Cambridge, UK.
Würzner
R
R
Hobart
M J
MJ
Lachmann
P J
PJ
D000116:Acetylgalactosamine / Q000276:immunology; D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D001826:Body Fluids / Q000276:immunology; D015938:Complement Membrane Attack Complex / Q000032:analysis; D003170:Complement Pathway, Alternative / Q000276:immunology*; D003165:Complement System Proteins / Q000032:analysis; D004443:Echinococcosis / Q000276:immunology / Q000662:veterinary*; D004446:Echinococcus / Q000187:drug effects / Q000276:immunology*; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010504:Periodic Acid / Q000494:pharmacology; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology*
D015938:Complement Membrane Attack Complex; D010504:Periodic Acid; C050974:complement C5b-6 complex; D003165:Complement System Proteins; C009288:metaperiodate; D010450:Endopeptidases; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D000116:Acetylgalactosamine