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immunization route resulted in high IFN- expression indicating a Th1 bias even though the IgG2a response was not very high (Fig

immunization route resulted in high IFN- expression indicating a Th1 bias even though the IgG2a response was not very high (Fig. cellulaire chez des souris BALB/c. Les souris ont t immunises avec le plasmide recombinant pDRIVE-EgAgB8/2 ou avec le plasmide vide pDRIVE en utilisant les voies dimmunisation suivantes : intramusculaire (i.m.), intranasale (i.n.) ou pidermale via un pistolet gnes (g.g.). Lanalyse de la production des anticorps et des cytokines a rvl que limmunisation par ADN par la voie i.m. a induit une rponse immunitaire de type T helper 1 (Th1) qui se Mouse monoclonal to Fibulin 5 caractrise par une haute expression du gnes IFN-et un faible rapport dIgG1/IgG2a (0,04). La voie i.n. a montr une expression modre dIFN-(AgB). which cycles between a carnivorous definitive host (mainly doggie) and herbivorous livestock intermediate hosts. Humans appear in the cycle as accidental host after ingesting food contaminated with parasite eggs (Eckert & Deplazes, 2004). CE affects the liver and the lungs mainly but other organs such as the spleen, pancreas, abdominal and pelvic cavities, musculoskeletal system, kidneys, heart and even the brain can be affected. The infection leads to morbidity and mortality worldwide and represents a significant hazard in most developing countries, where it impairs human health and considerable loss in animal production (Eckert & Deplazes, 2004). Studying the immunological profile of human patients with different stages of hydatid disease revealed that parasite survival is always supported with Th2 lymphocyte polarization of hosts immune system (Rigan infection has been attempted to modulate the immune system toward a Th1 protective response by the use of cytokine genes along with Eg95 (an oncospheral protein product) (Scheerlinck contamination is based on the fact that this egg infective stage of the parasite enters the intermediate host and humans through the oral or nasal ports of entry rendering the mucosal lining as the first line of contact with the parasite. Materials and Methods Preparation of crude sheep hydatid fluid Crude sheep hydatid fluid (CSHF) was Choline Fenofibrate aspirated from fertile hydatid cysts obtained from infected livers and lungs of sheep slaughtered at local abattoir in Irbid, Jordan. CSHF was centrifuged at 1,000??g for 15?min and the supernatant was dialyzed against phosphate buffered saline (PBS, pH 7.2) then filtered using a millipore filter (0.2?m), lyophilized using Edward, EF Modulyo lyophilizer (Crawley, United Kingdom) and stored at -20?C until use. Following reconstitution of the lyophilized material at 2?mg/mL with phosphate buffer saline (PBS) (pH 7.2), the protein concentration was determined using Bradford method (Bradford, 1976). Preparation of AgB Native AgB was prepared from a pool of CSHF based on the method of Oriol (1971) with some modifications. Briefly, 50 mL of CSHF were dialyzed against 0.005?M acetate buffer (pH 5) overnight at 4?C and centrifuged at 13,000??g for 30?min (Sigma, USA). The precipitate was dissolved in 5 mL of Choline Fenofibrate 0.2?M phosphate buffer (pH 8), autoclaved for 15?min?at 15 psi Choline Fenofibrate and 120?C and centrifuged at 13,000??g for 60?min. The supernatant made up of AgB was filtered by a millipore filter (0.2?m), aliquoted and stored at -20?C until use. Plasmid DNA preparation A recombinant plasmid pDRIVE-EgAgB8/2 was constructed as previously described (Al-Aghbar DH5 and purified using Wizard? Plus Maxipreps DNA Purification kit (Promega, USA) to remove possible endotoxin contamination. The concentration of each plasmid DNA was decided spectrophotometrically at 260?nm and stored at -20?C until used. Preparation of DNA gold microcarriers DNA-coated gold beads for Gene Gun immunization.