The Relationship Between FcγRⅡ Expression Level And The Immune State Of Chronic HBV Infection

Background and aims:The mechanism of persistent hepatitis B virus(HBV)infection and it induced immune tolerance has been a widely concerned topic.Our project used microarray to analyze the whole genome of PBMC that got from HBV infected human.We found that FcγRs had a significant difference in different phases of chronic hepatitis B.After that we performed PCR analysis to detect FcγRs m RNA expression in patients with HBV infection.The study showed that the inhibitory FcγRⅡb expression in immune tolerance period of CHB patients was significantly higher than that in healthy control group and immune clearance period group,and the activated FcγRⅡa expression in HBV immune clearance period group was significantly higher than that in healthy control group and HBV tolerance period group.Then we performed flow cytometry analysis to detect FcγRⅡ(CD32)expression on B cells and their subsets in different HBV infected patients.The study showed that CD3-CD19+CD32+B cells in patients with immune activated phase of chronic hepatitis B(CHB)were higher than that in patients with immune tolerance phase group and healthy control group.CD3-CD5 +CD32 + B cells in immune activated group was higher than that in healthy control group.These findings suggested that FcγRⅡb and FcγRⅡa may play an important role in the immune response of chronic HBV infection.The purpose of this paper is to explore FcγRⅡb and FcγRⅡa expression level in serum of chronic HBV infection patients and the difference of FcγRⅡb expression in liver tissue of patients with CHB to further verify the correlation between FcγRⅡb and FcγRⅡa expression and chronic HBV infection.Methods:We performed enzyme linked immunosorbent assay(ELISA)to detect the serum level of FcγRⅡb and FcγRⅡa of 119 patients with chronic HBV infection(including64 HBV carriers and 55 CHB patients)as well as 24 controls.The correlation between FcγRⅡb and FcγRⅡa expression level and clinical parameters such as AST、ALT was analyzed in CHB patients.Then we used immunohistochemistry to detect FcγRⅡb expression level in liver tissue of 14 CHB patients and 4 controls.The integrated optical density(IOD)values were measured by Image-Pro Plus 6.0 software to represent FcγRⅡb expression.The correlation between FcγRⅡb expression level and clinical parameters such as AST、ALT was analyzed.Results:1、The values of serum FcγRⅡb were 201.9±15.19ng/ml in healthy control group,183.9±33.47 ng/ml in HBV carrier group and 141.0±37.14 ng/ml in CHB group.Compared with healthy control group,there was no significant difference of serum FcγRⅡb level in HBV carrier group,while FcγRⅡb level in CHB patients was significantly lower(P < 0.001).The serum FcγRⅡb level in CHB patient group were significantly lower than in HBV carrier group(P < 0.001).Among all the HBV infected patients,the serum FcγRⅡb level of HBe Ag positive patients was statistically significantly lower than that of HBe Ag negative patients(P = 0.007).2、FcγRⅡb level had a negative correlation with AST(r=-0.3936,P=0.0063)and ALT(r=-0.3459,P=0.0097).There was no significant correlation between FcγRⅡb level and serum ALP、γ-GTP、albumin and HBV DNA.3、 The values of serum FcγRⅡa were 65.20±7.47 ng/ml in healthy control group,68.56±14.05 ng/ml in HBV carrier group and 94.35±29.50 ng/ml in CHB group.Compared with healthy control group,there was no significant difference of serum FcγRⅡa level in HBV carrier group,while FcγRⅡa level in CHB patients was significantly higher(P < 0.001).The serum FcγRⅡa level in CHB patient group were significantly lower than in HBV carrier group(P < 0.001).Among all the HBV infected patients,the serum FcγRⅡa level of HBe Ag positive patients was statistically significantly higher than that of HBe Ag negative patients(P = 0.009).4、FcγRⅡa level had a positive correlation with AST(r=0.5332,P<0.001)and ALT(r=0.4218,P=0.0013).There was no significant correlation between FcγRⅡa level and serum ALP,γ-GTP,albumin and HBV DNA.5、The FcγRⅡb expression was only found in liver sinusoidal endothelial cells(LSECs)The expression level of FcγRⅡb were 23696.08±3847.33 in control group、21392.93±7536.20 in mild CHB group、10287.25±2878.82 in mediate CHB group and 5214.78±1071.27 in severity CHB group.Compared with control group,there was no significant difference FcγRⅡb level in mild CHB group(P=0.84),while FcγRⅡb level in moderate CHB and severity CHB group were significantly lower(P=0.006,P<0.001,respectively).The FcγRⅡb expression level in severity CHB and moderate CHB group were also lower than in mild CHB group(P<0.001,P=0.018,respectively).6、The correlation analysis showed that FcγRⅡb had a negative relationship with AST(r=-0.688,P=0.0016)and ALT(r=-0.686,P=0.0017),and a positive relationship with platelets((r=0.6464,P=0.0038).There was no significant correlation between FcγRⅡb level and serum ALP,γ-GTP,albumin and HBV DNA.The comparison of pathological stage showed that FcγRⅡb had a significantly negative correlation with inflammation grade(r=-0.913,P<0.001)and fibrosis state(r=-0.875,P<0.001).It could be suggested that the decline of FcγRⅡb levels in the progression of chronic hepatitis correlates with the liver inflammation and fibrosis getting worse.Conclusion:1.This study verified our previous results of whole genome array techniques and Real time PCR as well as flow cytometry analysis in patients of different stage of HBV infection that FcγRⅡb and FcγRⅡa expression level had a correlation with chronic HBV infection.2.Inhibitory receptor FcγRⅡb level was negatively correlated with AST,ALT in CHB patients,however,activated receptor FcγRⅡa was positively correlated with AST,ALT.FcγRⅡb and FcγRⅡa may play a significant role in immune response of chronic HBV infection.3.The expression of FcγRⅡb in liver tissue was negatively correlated with AST and ALT while positively correlated with platelet level.The change of FcγRⅡb expression may influence the degree of liver inflammation and fibrosis in CHB patients.4.The expression levels of FC γ R Ⅱ B and FC γ R Ⅱ a are related to immune tolerance and immune activation in the process of chronic HBV infection,which can be used as immunological markers to distinguish the two immune states.