| Physalis alkekengi L. also called Calyx seu Fructus Physalis (CSFP) is a memberof the family of Solanacese and a herb of perennial root. Its calyx, whole herb androot can all be used as medicine. The whole herb of CSFP contains physanol A andB.Seedling, leaf and stem contain Physalin A, B and C alkaloid, citric acid and othertrace material that are necessary for human growth. In recent years, further researchon plant polysaccharide proves that many plant polysaccharides have functions asgood immune regulators.Polysaccharide can not only activate immunocytesefficiently, enhance the body’s immunity, but also have no side effects on normal cells,so it is considered to be an important biological response modifier. This paperselected water-soluble polysaccharide (WSP) from the stem of Physalis alkekengi L.to research into and explore its immune enhancement effect in DNA vaccine. Theresult showed that WSP had significant immune enhancement effect.WSP was chosen as adjuvant to immunize ICR male mice together with DNAvaccine that was constructed in our laboratory. It was administered by anterior tibialinjection,and supplemented by electroproration stimulation.The mice were immunizedintraperitoneally two times at a14-days interval.14days after the second injection,blood samples were collected from the eyes of the mice. Serum was collect and toexplore the immune enhancement effect of WSP, the following experiments wereconducted: Western Blot to detect the specific of antibody in the immuned miceserum; Detection of the titer of specific antibody by ELISA; Detection of the theamount of IL-2, IL-4, IL-10and IL-12concentrations in the serum by ELISA kit.Also mice were intentionally infect with sub-lethal Candida albicans by injectionthrough the tail vein to explore what effect WSP will on the mice immuned bypD-HSP90C.The results showed that pD-HSP90C can induce immune specific antibodies inmouse serum as a nucleic acid vaccine. WSP as a vaccine adjuvant immunizedtogether with the DNA vaccine in mice showed that its specific antibodies IgG, IgG1and IgG2b titers and IL-2, IL-4, IL-10and IL-12in serum were significantlyincreased compared to single injection of DNA vaccine groups. The group injected inthe tibialis anterior muscle with supplementary intramuscular electroporationstimulation indicated the specific antibody and cytokine levels also increased significantly compared to the groups that directly injected intramuscularly. Immuneprotection experiment results showed that, WSP immunized together with DNAvaccine mice groups whose renal tissue was cultured Candida albicans colonies weresignificantly less than the groups injected with DNA vaccine. The groups injectedthrough the tibialis anterior muscle with supplementary intramuscular electroporationstimulation had the number of colonies also significantly reduced compared to thegroups with intramuscular injection. Renal biopsy results of the analysis indicate that,the extent of the damage glomerular and tubular than was significantly lower in WSPtogether with DNA vaccine immunized mice groups than in the group immunizedwith DNA vaccine alone. The groups immunizes through the tibialis anterior musclesupplemented by intramuscular electroporation stimulation showed the extent ofdamage in the tubular and glomerular was significantly lower compared to the groupsof direct intramuscular injection.This study showed that WSP as a DNA vaccine adjuvant and electroporation asan immune method can significantly enhance Th1and Th2immune responsesignificantly in mice and in also mice immunized with DNA vaccine against systemicCandida albicans infection. These findings laid a solid foundation for the theapplications of WSP in the vaccine adjuvant field as well as the application ofelectroporation technique. |
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