Construction And Identification Of Novel Somatostatin DNA Vaccine Containing CpG Motifs And Study On It's Enhancement Mechanism And Immunological Effects

pES/2SS-CpG containing CpG motifs was constructed. A series of techniques were used, such as gene cloning, ELISA, RIA and regular chemical analysis. Using pES/2SS, pES/2SS-CpG, pES/2SS+pE-CpG immune mouse, we observed its effect on immune response and analyzed the effect on lymphocyte proliferation and the concentrations of GH and IGF-I. The whole experiment is to explore the use of CpG as an effective adjuvant.1 Construction and identification of somatostatin DNA vaccine pES/2SS-CpGWe designed two CpG-ODN fragments that containing 8 CpG motifs on the related reference data.The two ODN were denatured at 95 ℃ and inserted into pES/2SS by selecting appropriate site. After this, we identified plasmid sequence by restricted endonucleases digestion and sequencing. Our aim is to promote somatostatin DNA vaccine immune effect by molecule adjuvant.2 Immune response and affecting factors of SS gene vaccine containing CpG motifsA total of 40 rats were randomly divided into 4 groups (n=10) including three experimental groups (T1～T3) and one control group(C). 20μl procaine hydrochloride (0.5%) was injected 24 h before immunization. T1-T3, namely the groups of plasmid pES/2SS(T1), pES/2SS-CpG (T2) and pES/2SS+pE-CpG(T3) at 20μg/20μl dosage, were injected at the same site as the pretreatment. The control group (C) was injected with 20μl of saline. The concentrations of GH and IGF-I in the plasma were detected by RIA technology. Every sample was diluted with 1:50 and the P/N value was measured by indirect ELISA and go to the fundamental that the P/N value more than 2 point is positive.The antibody against SS level and the spleen lymphocyte proliferation chance were detected by ELISA and MTT colorimetric assay respectively. The results showed that Ratio of positive antibody against SS of T1-T3 ranged between 30.0% and 40.0%. All the three experiment groups were the most significantly higher than that of the control (p<0.01). The max value of P/N was about 3.76. The best immune activity of plasmid containing CpG was 40% positive, which was not significantly higher than pES/2SS(P>0.05). The lymphocyte proliferation of the three experiment groups were the most significantly higher...