[Repelling property of the methalonic and hexanic extracts of Sambucus ebulus L. against the Culex pipiens: in-vitro study]

Background and Objective: The anti inflammatory, analgesic, antioxidant of Sambucus ebulus L. have been reported in several studies. This study was done to assess the repelling property of the methalonic and hexanic extracts of Sambucus ebulus L. against the Culex pipiens

Methods: In this experimental study, Sambucus ebulus L. collected from the natural inhabitants of Mazandran province in northern Iran. Methalonic and hexanic extraction were provided from the leaf and fruit of Sambucus ebulus L. Concentration of 50 mg/kg, 100 mg/kg and 250 mg/kg was prepared. 0.4 ml of the extract prepared and was spreed on the albino skin area of 4×6 cm2. After 30 minutes the number of the mosquito [Culex pipiens] bites on the skin was recorded. N, Ndiethyl-3 methyl benzamide was considered as positive control

Results: The highest repelling property of the Sambucus ebulus L. belonged to the concentration of 250 mg/kg of leaf and fruit extraction. The highest repelling effect was 80% and 66.8% for leaf methalonic and hexanic extract, respectively. The highest repelling effect was 84% and 72% for fruit metalonic and hexanic extract, respectively [P<0.05]

Conclusion: The methalonic extract of Sambucus ebulus L. had higher repelling efficiency compared to the hexanic extract. The fruit extract also had better effect than the leaf extract

[Study the capacity of recombinant HCV core+1 protein to induce immune responses in combination with different adjuvants]

Hepatitis C virus [HCV] genome contains a large open reading frame coding for a polyprotein that is cleaved into ten proteins. Recently, a new protein, named core+1, has been described to be expressed through a ribosomal frame shift within the capsid-encoding sequence. To address these possibilities, core+1 was produced in E.coli and the purified protein was evaluated for the immunological properties. The core+1 corresponding nucleotide sequence was created by PCR-based induction of a +1 frame shift mutation within the core gene template. The amplicon was cloned into the pET-24a vector and expressed in E.coli host. The expressed protein was purified under denaturing condition and after refolding steps was characterized by SDS-PAGE and Western Blotting. The immunization potential of core+1 with various adjuvant [Freunds [C/IFA], Montanide ISA 206 and IMS 1312, pluronic acid [F127] and imiquimod [IMIQ] was assessed in Balb/c mice. ELISA-based assays were used to analyze the humoral immune responses. The yield of E.coli-derived core+1 was 5 mg/ L of culture media and antigenicity of this protein was confirmed by western blotting. All the core+1/adjuvant formulations significantly developed the anti-core+1 IgG responses in the immunized mice but C/IFA and ISA206 elicited the highest antibody titers. ISA 206 and IMS 1312 formulation of core+1 induced strong Th1/Th2 responses. Our results indicated that core+1 formulation with various adjuvants may elicit the different immune response profiles [Th1/Th2]. Thus core+1 might be a potential component of future HCV vaccine too

[Designing and producing polytope DNA vaccine containing HBsAg gene for the induction of protective immunity against hepatitis C]

Background: considering the immunosuppressive effects and prevalent mutations in some HCV antigens, induction of CD8[+] T cell responses is focused on conserved and critical epitopes which as a multi-epitope vaccine can prevent the chronic nature of the disease

Materials and Methods: two immunodominant HLA-A2-restricted human epitopes [E2[614- 622] and NS3[1406-1415]] and two H-2[d]-restricted mouse epitopes [core[132-142] and E2[405-414]] were designed in a sequential tandem, predicted by immunoinformatic analyses. Following the synthesis, related nucleotide sequence was cloned into the pcDNA3.1 vector with and without the fusion of hepatitis B surface antigen [HBsAg]. Two constructed plasmids [pcDNA3.1.HPOL and pcDNA3.1.POL, respectively] were evaluated for the protein expression and secretion in Cos-7 cell line. After the vaccination of BALB/c mice [n=6 in each group] with different DNA and peptide immunization regimens, CD8[+] T cell activity as well as the type and protective potency of the induced responses were evaluated

Results: despite the induction of epitope-specific responses in pcDNA3.1.POL injected mice, the group immunized with pcDNA3.1.HPOL indicated a significant increase in the number and activity of CD8[+] T cells [P<0.05]. Peptide boosting of this group [formulated in two human-compatible adjuvant] still led to the more activation of CD8[+] cells, induction of Th1 response and the inhibition of tumor model growth [P<0.05]

Conclusion: fusion of HBsAg as a particle-forming sequence and the source of helper epitopes along the DNA-prime/peptide-boosting immunization regimen are proposed as two promising strategies to improve the CTL multi-epitope vaccines against HCV