Experimental Research On The Effects Of Angelica Polysaccharide On Dendritic Cells In HBV Trans-genic Mice

Backgroud: The Hepatitis B virus(HBV) is an important cause of chronic Hepatitis B(CHB), cirrholosis of liver and hepato celluar carcinoma..In our country, the patients of ASC are about 1.2 millions, CHB are about 28 millions.But the status of clinical treatments are not satisfacting. Studies find that,as an important antigen-presenting cell, the functional impairment of dendritic cell(DC) mignt be an important cause of HBV chronic infection . Angelica polysaccharide(APS) has a role of strengthing immuno functions of body by inducing body to produce many kinds of cytokines(CKs).But what effects do APS have on the DC of CHB and wether it strengthes the function of anti-virus by inducing the body to produce CKs which can stimulate DC proliferation and mature havn't been reported.Objective: To study the effects of Angelica polysaccharide(APS) on the functional status (such as the mature status , the capacity of expressing CD and stimulating lymphocyte proliferation and excretion of CKs) and the immuno capability of anti-virus of DC by experiments.To investigate the theory of effects initially.Methods monocytes of HBV trans-genic mice treated with and without APS were isolated from spleen and incubated with rmGM-CSF and rm-IL-4 to induce DC generation. The morphotype of DC was identified by microscope.The phenotype of DC(CD₈₀,CD₈₆) was characterized by flow analysis cytometry.The capacity of stimulating reaction of allogenic T lymphocytes was detected by MTT assay and stimulating index(SI) were calculated..The concentration of cytokines ((IL-12,r-IFN) in supernate and HBsAg, HbeAg in serum of mice was tested by ELISA.A11 these results of HBV trans-genic mice were compared with normal mice.Results: monocytes of HBV trans-genic mice were isolated from spleen and incubated with rmGM-CSF and rm-IL-4 to induce DC generation.Some fusiform or clavillose cells with sentus enation can be seen in the microscope.In the tele-microscope we can see the surface of cells had reductus and branch enation and their cellular nuclei were irregular.Detected by flow analysis cytometry, the expression rates of CDson CDg6 of DC in the mice treated with APS were 6.1 ± 1.21% s 12.3 ± 3.07%,but the expression rates of control were 2.1 ± 0.12% s 2.63 + 0.02% ,the expression rates of CDx6 of DC in treated mice was higher than control, the capacity of presenting antigen of DC in treated mice increased. The expression rates of CD8o-CDg6 in normal mice were 20.7 ± 1.94%> 31.2± 1.82%,higher than HBV tans-genic mice.As DC mixed with lymphocyte on the concentration of 1:10.1:50.1:100 in MLR.SI of DC in mice treated with APS were 1.50 ± 0.09 s 1.08 ± 0.08 s 0.81 ± 0.10,SI of controls were 0.87±0.1k 0.74±0.04s 0.47±0.27.SI of DC in treated group was increased compared with that of control when DC and lymphocyte were mixed on the concentration of 1:10.1:50.The capacity of stimulating lymphocyte proliferation of DC in treated mice increased. SI of normal mice were 2.84 ± 0.32> 2.30 ± 0.19s 2.22 ± 0.15,higher than HBV tans-genic mice.The amounts of cytokines(r-IFN> IL-12) in MLR of treated mice were 59.51 ±2.18 pg/ml s 247.64± 31.59 pg/ml, the control were 53.44+ 1.37 pg/ml, 164.01 ± 12.75 pg/ml. which were measured by ELISA. The amounts of r-IFNs IL-12 in MLR of treated mice were higher than those of controle. The two CKs measured in normal mice were 106.91 ±4.73, 482.64 + 43.3 pg/ml, higher than HBV tans-genic miceIn the same time, when we measured the concentration of HBsAg in serum of mice by ELISA, we found the mean of P/N in treated mice and control were respectively 25.65 ± 4.89s 37.76 ± 6.90.The concentration of HBsAg in treated mice was lower than the control.Conclusion: Inducing and culturing the monocytes from spleen of HBV trans-genic mice with rmGM-CSF , rm-IL-4 and RPM11640 culture can get mature DC; The immuno function of DC in HBV tans-genic mice were impair, which was perhaps the most important cause of HBV chronic infection; APS can promote the mature of DC in HBV trans-genic mice, increase the expression of CDS6 on the DC and the capacity of stimulating lymphocyte proliferation and excretion oflL-12s r-IFN. It can strengthen DC s functions of presenting antigen and stimulate cellular immunological response .As a result, the concentration of HBsAg in HBV trans-genic mice decreased on some extent. It maybeplay a role in the anti-virus immunological response of HBV trans-genic mice.