ABSTRACT
Abstract INTRODUCTION: High percentages of structural identity and cross-immunoreactivity have been reported between potato apyrase and Schistosoma mansoni ATP diphosphohydrolase (SmATPDases) isoforms, showing the existence of particular epitopes shared between these proteins. METHODS: Potato apyrase was employed using ELISA, western blot, and mouse immunization methods to verify IgE reactivity. RESULTS: Most of the schistosomiasis patient's (75%) serum was seropositive for potato apyrase and this protein was recognized using western blotting, suggesting that parasite and plant proteins share IgE-binding epitopes. C57BL/6 mice immunized with potato apyrase showed increased IgE antibody production. CONCLUSIONS: Potato apyrase and SmATPDases have IgE-binding epitopes.
Subject(s)
Animals , Female , Apyrase/immunology , Schistosoma mansoni/immunology , Schistosomiasis mansoni/immunology , Solanum tuberosum/enzymology , Immunoglobulin E/immunology , Antibodies, Helminth/immunology , Epitopes/immunology , Enzyme-Linked Immunosorbent Assay , Blotting, Western , Cross Reactions , Mice, Inbred C57BLABSTRACT
A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.
Subject(s)
Animals , Male , Mice , Antibodies, Helminth/immunology , Antigens, Helminth/immunology , Apyrase/immunology , Schistosoma mansoni/immunology , Blotting, Western , Cross Reactions , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Egg Proteins/immunology , Immunohistochemistry , Schistosoma mansoni/enzymologyABSTRACT
In this paper, we showed for the first time that the conserved domains within Schistosoma mansoni ATP diphosphohydrolase isoforms, shared with potato apyrase, possess epitopes for the IgG1 and IgG4 subtypes, as 24 (80 percent) of the 30 schistosomiasis patients were seropositive for this vegetable protein. The analyses for each patient cured (n = 14) after treatment (AT) with praziquantel revealed variable IgG1 and IgG4 reactivity against potato apyrase. Different antigenic epitopes shared between the vegetable and parasite proteins could be involved in susceptibility or resistance to S. mansoni AT with praziquantel and these possibilities should be explored.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Animals , Child , Child, Preschool , Humans , Middle Aged , Young Adult , Antibodies, Helminth/immunology , Apyrase/immunology , Immunoglobulin G/immunology , Schistosoma mansoni/immunology , Schistosomiasis mansoni/immunology , Solanum tuberosum/enzymology , Anthelmintics , Cross Reactions , Praziquantel , Schistosomiasis mansoniABSTRACT
Nesta tese caracterizamos uma nova enzima na superfície do tegumento do Schistosma mansoni, uma ATP difosfohidrolase. Controles apropriados mostraram que as atividades ATPásica, ADPásica, pirofosfatásica e adenilato quinásica possivelmente contaminantes não são significativamente importantes. Foi mostrado também dependência para íons divalentes e um provável mecanismo de hidrólise sequencial. Experimentos anticorpos com sugerem que sua massa molecular esteja em torno de 50 KDa. É uma ecto-enzima, foi o que comprovado citoquímica por em microscopia eletrônica e pela capacidade do esquistossomo de hidrolisar ATP ou ADP adicionados no lado externo de vermes intactos. Parece ser sintetizada pelo próprio verme, já que esquistossômulos obtidos in vitro apresentaram atividades ATPásica ADPásica e nas condições mesmas usadas para medidas tegumento de vermes adultos. Esta ecto-enzima pode estar envolvida nos mecanismos de escape do parasita ao sistema hemostático e/ou imune do hospedeiro, bloqueando os sistemas que usam ADP e/ou ATP como intermediários.