| Polysaccharides are important biological macromolecules in organisms.Polysaccharides have many effects,such as regulating immunity,anti-virus and so on.The purpose of this experiment is to strengthen the bioavailability of acidic camel thorn polysaccharide,prepare a vaccine adjuvant with good immune enhancement and long-term effect,provide a good vaccine adjuvant for the prevention of avian influenza and livestock infectious diseases,and provide a research method for the effective use of local medicinal materials.In this experiment,acid Alhagi sparsifolia shap polysaccharide(aASP)was extracted by response surface methodology,and the polysaccharide was purified and identified;AASP in vivo and in vitro enhanced immunity test,preparation of aASP loaded PLGA nanoemulsion(aASP-PPAS)for characterization and detection.aASP-PPAS was used as the adjuvant of the H9N2 nano vaccine to immunize chicks for adjuvant activity detection.The main results are as follows:1.The optimum extraction conditions were as follows: the extraction temperature was 95 ℃,the extraction time was 2.1 h,the solid-liquid ratio was 1:26,and the yield was 1.645%;2.Crude polysaccharides were purified by gel fiber DEAE-52 and Sephadex G-100 column,and purified aASP was obtained.In monosaccharide group,the polysaccharide mainly contains arabinose,galactose,mannose and rhamnose;The content test showed that the content of neutral polysaccharide was81.8 ± 1.2%,the content of aldehyde acid was 17.34 ± 0.75%,and the content of protein was 0.54 ± 0.08%.The test results showed that the polysaccharide of camel thorn was acidic.3.Crude polysaccharides were purified by gel fiber DEAE-52 and Sephadex G-100 column,and purified aASP was obtained.In the monosaccharide group,the polysaccharide mainly contains arabinose,galactose,mannose,and rhamnose;The content test showed that the content of neutral polysaccharide was81.8 ± 1.2%,the content of aldehyde acid was 17.34 ± 0.75%,and the content of protein was 0.54 ± 0.08%.The test results showed that the polysaccharide of camel thorn was acidic.4.aASP can enhance the proliferation of chicken spleen lymphocytes and promote IL-4,IL-6,and IFN-γ It can promote the production of the germinal spleen center and has a good immune effect.5.aASP-PPAS/H9N2 had a good sustained-release effect within 28 days,no precipitation within 64 days,and good stability.The antigen loading rate of aASP-PPAS /H9N2 was 64.7 ± 3.34%.AASP PPAS was spherical,and the surface was strawberry under a freeze scanning electron microscope.6.aASP-PPAS/H9N2 could stimulate the body to produce a high HI titer and IgG.There was no significant difference between aASP-PPAS/H9N2 and oil/H9N2(P > 0.05).aASP-PPAS/H9N2 could significantly stimulate and promote IL-4,IL-6,and TNF-α and IFN-γ Secretion of. |
