Regulation Of ?-Glucan On Cellular Immune Of Patients With Chronic Hepatitis B Infection

Objective:HBV(hepatitis B virus,HBV)is one of the main viruses affect human health.HBV persistent infection in the body leads to a host of CHB.CHB is gradually deteriorating,leading to formation of liver cirrhosis and liver cancer,therefore,prevention and treatment of CHB become an important medical subject.Peripheral immune response is an important place,in addition to having direct macrophage phagocytosis,clearance outside the role of PBMC produced such as interferon,IL-12,etc.And they also play an important role in immune regulation.DCs are now known to most powerful antigen-presenting cells,which also play an important role in the process.They activate naive T cells to stimulate the initial immune response.Dextran regulate immune and regulate metabolism and so on.It is an immune-most active and most easily absorbed by the body non-specific immunomodulator.In this study,by the method of vitro cell culture studies of patients with CHB DCs and PBMCs,using FACS affecting the detection of?-glucan on DC phenotype and PBMC's,explore the pathogenesis of CHB patients,?-glucan role in immune regulation.It provides an important scientific basis for the treatment of chronic hepatitis B immune cells and the development of new vaccines against CHB.Methods:1?Collecting 10 healthy volunteers by density gradient centrifugation of peripheral blood mononuclear cells,using serum-free media and serum-free media respectively to do DC cell culture,and flowing cytometric examination the expression level of CD11 and CD86 in the surface of DCs.2?Collecting 30 patients with CHB,12 cases healthy controls in the fifth hospital of Shijiazhuang.Gathering the peripheral blood of 8 mL,using EDTA anticoagulant.Lymphocyte separation medium separation by density gradient centrifugation method mononuclear cells,and used to cultivate PBMCs and DCs.Break them up into there groups,just as the group of no stimulation,the group of LPS and the group of?-glucan.In the course of training,use an inverted microscope cell size,shape and growth.3?Using APC-CD11c,PE-Cy7 CD86,FITC CD80 labeling DCs,each of them with 2?L.Using APC-CD11c,PE-Cy7-CD3,FITC-CD4,PE-CD8a and Percp-CD56 labeling PBMCs,to detect the expression of each cell by FACS.4?Collecting and sorting all data and applications SPSS19.0 software for statistical analysis.Results:1?Serum culture medium,serum-free culture medium DC cells,there have no difference in cell morphology and have a little difference in the form of the number,the expression of CD11c and CD86 in the surface of DCs was detected by flow cytometry between the two groups,CD11c expression differences,with statistical significance(P<0.05),Serum medium group slightly higher than serum-free media group,but little difference,CD86~+expression was no difference(P>0.05).2?In the experimenting of DCs culture and stimulation.Observed by inverted microscope,DC cells in patients with chronic hepatitis B compared with healthy cells DC,the density is low.Use of LPS,?-glucan stimulates after 24 h,compared with no stimulation,two groups of cell growth,cell body is bigger,the typical dendritic protuberance;LPS group cells mature cells early in the beta glucan group.3?The expression of CD11c and CD86 in the surface of DCs in patients with chronic hepatitis B and healthy cells DCs.The finding showed that peripheral blood cell surface DC healthy subjects and patients with chronic hepatitis B control group,LPS group and?-glucan group CD11c~+expression was not statistically significant(P>0.05);Healthy volunteers and in patients with DC cell surface CD80~+?CD86~+between the two groups was statistically significant(P<0.05);Healthy volunteers CD80~+,CD86~+expression were higher than HBV patients.Given different treatment,no stimulation group,LPS group,?-glucan group was statistically significant(P<0.05),compared with three groups,expression of CD80~+?CD86~+were no stimulation group<?-glucan group<LPS group.4?In the experimenting of DCs culture and stimulation.Observed by inverted microscope,join the stimulation training after 2 d,no stimulation group,LPS group,?-glucan cells morphologically had no differeence.5?Flow cytometry,each group PBMC CD11c~+cells expressed,the control group compared with the case group,no significant difference(P>0.05);Control group,LPS group,?-glucan pairwise comparison,was not statistically significant(P>0.05).Unstimulated group,LPS group,?-glucan group in CHB patients group and healthy controls group,the expression of CD3~+CD4~+have no significant difference(P>0.05),the expression of CD3~+,CD4~+quantity,no stimulation group<?-glucan group<LPS group.CHB patients group and healthy controls group in three groups of pairwise comparisons was statistically significant(P<0.05),Expression levels of CD3~+CD4~+were no stimulation group<?-glucan group<LPS group.The expression of CD3~+CD8,healthy voulunteers in comparison with patients,the two groups was statistically significant(P<0.05).Healthy volunteers in no stimulation group??-glucan group?LPS group group>these there groups of patients,The group of no stimulation group??-glucan group?LPS group group in CHB patients and healthy group comparisons was statistically significant(P<0.05),Expression levels of CD3~+CD8~+were no stimulation group<?-glucan group<LPS group.The expression of CD3~-CD56~+,healthy voulunteers in comparison with patients,the two groups were statistically significant(P<0.05).Healthy volunteers in no stimulation group??-glucan group?LPS group group>these there groups of patients,The group of no stimulation group??-glucan group?LPS group group in CHB patients and healthy group comparisons were statistically significant(P<0.05),Expression levels of CD3~-CD56~+were no stimulation group<?-glucan group<LPS group.The expression of CD3~+CD56~+,healthy voulunteers in comparison with patients,the two groups were statistically significant(P<0.05).Healthy volunteers in no stimulation group??-glucan group?LPS group group>these there groups of patients,The group of no stimulation group??-glucan group?LPS group group in CHB patients and healthy group comparisons was statistically significant(P<0.05),Expression levels of CD3~+CD56~+were no stimulation group<?-glucan group<LPS group.Conclusions:1?Low immune function of patients with CHB is closely related to the low maturation status of DCs.?-glucan and LPS can promote DC proliferation and induce cell maturation.2?Under the stimulus LPS and?-glucan,the maturation of DCs has no changed,the antigen presenting capacity of DCs has been improved.3?Patients with CHB compared with healthy controls,the number of Th cells has no change.The number of CTL cells,NK cells and NKT cells in the patients with CHB are less than the number of all healthy controls,thereby resulting in slow immune tolerance of hepatitis B patients.4?LPS,?-glucans stimulated PBMC cells,increased expression of CTL cells and NK cells,and NKT cells decreased.5??-glucan have functions in immune regulation and can improve the state of immune tolerance in patients with CHB.It can offer a new method to treatment of CHB...