|
Hepatitis B is a serious threat to human health, disease, if not treated, can cause cirrhosis, liver cancer and other malignant diseases, and even death. Vaccination is an effective means for the prevention of hepatitis B, but for the treatment of hepatitis B, especially in the treatment of chronic hepatitis B has little effect - traditional vaccines induce humoral immune response, the cellular immune response is weak cellular immune Clear body within the virus plays a central role. Vaccines do this research and development can lead to high levels of cellular immune response to vaccine research hotspot. Therapeutic vaccine can induce strong cellular immune favored by researchers, the DNA vaccine is a promising therapeutic vaccine. Although DNA vaccines can induce the ideal cell and humoral immune responses, but still there is a certain lack of, i.e. in the instance of larger animals and people induced a weak immune response, the immune effect is not satisfactory. To this end, researchers seeking to overcome this shortcoming through different channels. Adjuvants can enhance antigen-induced immune response and effectively extend the effective time, it is considered to be the ideal way to an enhanced immune effects. Astragalus polysaccharide (APS) is the main active ingredient of traditional Chinese medicine Astragalus has a good immune enhancement, can promote the body's CD4 T lymphocyte proliferation and IL-2 and IFN-γ levels, promote the proliferation of spleen cells and B lymphocytes, can promote dendritic cell maturation, enhance the antigen-presenting ability, and activated macrophages. This study astragalus polysaccharide as an adjuvant with recombinant pcDS2 plasmid with mice immunized to evaluate its impact on the immune response in mice by detecting the level of humoral and cellular immune responses; study the role of dendritic cell maturation, set forth in order to affect the immune response mechanism. This study selected 50 6-8 week-old healthy female Kunming white mice were randomly divided into five groups of 10 each. Each test group treatment: blank control, a single injection of the plasmid vector, the only injection the reorganization of pcDS2 plasmid, the the injection pcDS2 plasmid and APS single injection of APS, and in turn named Naive group the pcDNA3 group the, pcDS2 group, pcDS2 APS group and APS group. Primary immunization time is set to 0, the early Free after 14 and 28 days each booster immunization, T cell proliferation was measured surface molecules and cytokines serum determination of antibody content acquisition in the first 35 days, the spleen cell suspension was prepared The expression levels of CTL killing capacity and signaling pathways of NF-κB and the expression level of p38 MAPK. The results obtained in this study are as follows: to the Naive groups, pcDNA3 group and APS group antibody levels were negative the the APS group antibody content of pcDS2 (8464.8ng/ml) significantly the to higher than pcDS2 group (3319.6ng/ml). CD4 T cells expressing IL-4 levels (with the proportion of the total CD4 T cells express IL-4 CD4 T cells to represent) the Naive group, pcDNA3 group the, pcDS2 group, pcDS2 APS group, APS group were 0.64%, 0.97%, 1.06%, 1.42%, 0.90%, which pcDS2 APS group was significantly higher than the other groups, APS can induce the immune response to Th2-type development. The above results show that the APS as an adjuvant can enhance the humoral immune response in mice. T cell proliferative stimulation index the Naive group of, pcDNA3 group the, pcDS2 group, pcDS2 APS group, APS group 1.4,1.4,2.4,3.2 and 1.7, respectively. pcDS2 APS group was significantly higher than the control group, showed the best proliferative capacity. The CD4'T cell IL-2 expression levels Naive group, pcDNA3 group, pcDS2 group, pcDS2 APS group and APS group were 0.26%, 0.43%, 0.58%, 3.42% and 0.36%, respectively. IL-2 expression levels of pcDS2 APS group is the highest, significantly higher than the other groups, APS has good promote T cell proliferation. A CD4'T cells and CD8'T cell of IFN-y expression level of each group (order op) were 0.10% and 0.21%, O.10% and 0.27%, 0.11% and 0.30%, 0.23% and 0.43%, as well as O.14% and 0.27% respectively. pcDS2 APS group, the expression level of a significant difference compared with other groups, APS have the ability to induce Thl type immune response. The highest level (56.5%) in vivo CTL killing experiments pcDS2 APS kill target cells also significantly higher than the control group (the Naive group 2%, pcDNA3 group 5.9%, pcDS2 group 27.6%, APS Group 3.9%) These results suggest that APS as an adjuvant can enhance the cellular immune response in mice. Costimulatory the signal molecules CD40, CD80, CD86 expression levels were Naive group 0.69%, 0.57%, 0.49%, pcDNA3 group, 0.66%, 0.51%, 0.55%, pcDS2 group, 0.92%, 0.77%, 0.88%, pcDS2 APS group 1.15%, 1.10%, 1.23%, APS0.78%, 0.64%, 0.78%, which pcDS2 APS group of three molecules expression were higher than the control group showed the strongest provide a second signal for T cell activation ability; DCMHCI and class II molecules and chemokine receptor CCR7 of expression is also pcDS2 the APS group the highest expression levels. In addition, pcDS2 APS group in the molecules involved in DC maturation signaling pathway NF-κB and the expression level of p38 MAPK is also higher than the other test group. The above results show that the APS promoting the maturation of the dendritic cells of the immunized mice. In summary, APS as HBV DNA vaccine adjuvant can promote DC maturation immunized mice, to enhance Th activity, to enhance the cellular and humoral immune response. This study provide a new way to enhance the effect of HBV DNA vaccine adjuvant potential at the same time show that APS, demonstrating good prospect.
|