IMMUNOPROTECTION AND DIAGNOSTIC POTENTIAL OF SIGN
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The 14-3-3 protein is a key player in signal transduction processes in various species of animals and plants. Here, we cloned and expressed the 14-3-3 of Schistosoma japonicum (Sj14-3-3, Chinese mainland strain) in pET28a/E.coli. And the protein can be strongly recognized by polyclonal anti-14-3-3εand sera from animals infected with S. japonicum. Monoclonal antibodies against Sj14-3-3 were raised using a highly specific, affinity-purified protein. The antibodies preparation by ammonium sulfate precipitation was employed for the localization of the native 14-3-3 protein in the parasite by immunofluorescence microscopy. The results demonstrate a wide distribution of this protein in tegument, subtegument, muscle and parenchyma of male, female adult parasites and 15-day-old schistosomulum. The potential use of this protein or its DNA as a vaccine candidate against infection with S. japonicum in BALB/c mice and the recombinant Sj14-3-3 (rSj14-3-3) antigen for diagnosis of schistosomiasis was evaluated. Challenge infection with S. japonicum, immunization with rSj14-3-3, rSjGST, and rSj14-3-3+rSjGST proteins using Freund’s adjuvant or Mycobacterium tuberculosis (Mtb) low molecular peptide as adjuvant respectively, led to 26.0-32.2% protection, as determined by reduction of adult worm burden; 50.4-58.6% protection as determined by reduction of the eggs in liver tissue. The mice were directly inoculated with rSj14-3-3 and rSj14-3-3+rSjGST cDNAs through quadriceps femoris muscle for immunization. As the consequence, the worm reduction was found to be 34.20% in rSj14-3-3 group, 32.40%in rSj14-3-3+rSjGST group; the number of eggs in liver tissue was reduced by 50.74% and 55.23%, respectively. Indirect ELISA suggested that the rSj14-3-3 antigen is highly sensitive and specific for the diagnosis of schitosomiasis.
The 14-3-3 protein is a key player in signal transduction processes in various species of animals and plants. Here, we cloned and expressed the 14-3-3 of Schistosoma japonicum (Sj14-3-3, Chinese mainland strain) in pET28a/E.coli. And the protein can be strongly recognized by polyclonal anti-14-3-3εand sera from animals infected with S. japonicum. Monoclonal antibodies against Sj14-3-3 were raised using a highly specific, affinity-purified protein. The antibodies preparation by ammonium sulfate precipitation was employed for the localization of the native 14-3-3 protein in the parasite by immunofluorescence microscopy. The results demonstrate a wide distribution of this protein in tegument, subtegument, muscle and parenchyma of male, female adult parasites and 15-day-old schistosomulum. The potential use of this protein or its DNA as a vaccine candidate against infection with S. japonicum in BALB/c mice and the recombinant Sj14-3-3 (rSj14-3-3) antigen for diagnosis of schistosomiasis was evaluated. Challenge infection with S. japonicum, immunization with rSj14-3-3, rSjGST, and rSj14-3-3+rSjGST proteins using Freund’s adjuvant or Mycobacterium tuberculosis (Mtb) low molecular peptide as adjuvant respectively, led to 26.0-32.2% protection, as determined by reduction of adult worm burden; 50.4-58.6% protection as determined by reduction of the eggs in liver tissue. The mice were directly inoculated with rSj14-3-3 and rSj14-3-3+rSjGST cDNAs through quadriceps femoris muscle for immunization. As the consequence, the worm reduction was found to be 34.20% in rSj14-3-3 group, 32.40%in rSj14-3-3+rSjGST group; the number of eggs in liver tissue was reduced by 50.74% and 55.23%, respectively. Indirect ELISA suggested that the rSj14-3-3 antigen is highly sensitive and specific for the diagnosis of schitosomiasis.
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