con A purified hydatid glycoprotein fraction effectively diagnoses human hydatidosis

Diagnosis and quantification of Echinococcus granulosus in-fection in man and animal hosts are centralized to feasible con-trol this study included 93 serum sample, 25 sure positive hydatid cases confirmed surgically, 7 suspected cases diagnosed by indirect haemagglutination IHA and 41 cases other parasitic infection [15 S. mansoni, 8 fasciola, 7 Ascaris, 5 H. nana and 6 Ancylostoma] diagnosed by microscopic examination and were negative by ELISA and/or IHA for anti-hydatid antibody. Twen-ty negative serum samples served as healthy controls. Six types of hydatid fluid antigens [crude, host-free and Con-A purified] of human and camel origin were subjected to electrophoretic separ-ation [SDS-PAGE] and immunoblotting [EITB]. The anti-hydat-id IgG was detected in sera of the different groups for evalua-tion of sensitivity, specificity and diagnostic efficacy each type of antigens. Detection of circulating hydatid antigen [CAg] was performed using anti rabbit hyperimmune sera raised again-st Con-A purified either human or camel hydatid antigen. SDS-PGE revealed several bands ranging from 55-185- kDa with 10kD band shared by all antigens. The specific bands revealed by EITB for Con-A purified camel and human antigens were at 80, 110, 55, 110 kDa respectively. ELISA highest sensitivity [96.9%] was by using host-free Con-A purified glycoprotein fraction of human hydatied antigen. Highest specificity [98.4%] was reco-rded upon use of either Con-A purified camel or human antigen with 94.5% and 97.7 and diagnostic efficacy respectively. Detection of circulating antigen by polyclonal antibodies against Con-A purified human hydatid antigen revealed 91.8%specificity.

Splenic cytokine mRNA expression in mice intravenously injected ith schistosoma mansoni 26-kDa glutathione S-transferase or soluble egg antigen before infection with schistosoma mansoni cercariae

This work was designed to assess the changes in hepatic granuloma formation associated with hyporesponsiveness to schistosomal egg antigen [SEA] or S.mansoni-26GST[Sm-26GST]. Multiple small doses of SEA [10 micro g x 4] or Sm-26GST [I micro g x 4] were injected intravenously [i.v.] into C57B1/6 mice 7 days prior to cercarial exposure. Mice were sacrificed 6 and 8 weeks post-infection, hepatic granuloma diameter was evaluated and mRNA expression of splenic cytokines IFN- alpha, IL-2, IL-4, IL-10 and IL-12 was estimated using the semiquantitative reverse transcription-polymerase chain reaction [RT-PCR]. Compared to infected controls, both SEA and Sm-26GST-treated groups showed significant decrease in hepatic granuloma diameter 8 wks post infection [p.i.] associated with lowered level of IFN- alpha and IL-4 mRNA; while the levels of both IL-10 and IL-12 were higher than that of infected controls. The present data show that Sm-26GST has an antipathology effect similar to SEA if it is used by i.v. route. The predominant cytokine involved in this hyporesponsiveness is IL- 10 and to a lesser extent IL- 12

Identification and characterization of specific hydatid antigen fraction [s]

A specific hydatid antigen was prepared in this study from Echinococcus granulosus cyst in livers and lungs of camels. Elimination of host "camel" protein from crude hydatid fluid was achieved by two methods: Salting out using ammonium sulfate precipitation method and immunoaffinity purification using coupled anticamel antibody to cyanogenbromide activated sepharose 4B gel. Testing the prepared hydatid antigen against anticamel serum, using immunodiffusion method, indicated that the affinity purified hydatid antigen was almost completely purified from camel protein. Characterization of the affinity purified hydatid antigen, using immunoelectrophoresis, showed positive arc 5 precipitation when tested against known positive antihydatid sera. Further characterization with gradient gel electrophoresis, showed with silver stain that the dominant and most consistently demonstrable proteins occurred as a complex in the 52/62 kDa region. Strong reaction with the 52/62 kDa complex was consistently observed when the affinity purified hydatid antigen was probed with known positive reference antihydatid sera. The identified hydatid antigen fraction[s] with 52/62 kDa complex can provide promising noninvasive parameter for diagnosis of hydatidosis