| Background and aims:To identity immuno logical mechanisms that govern distinct clinical phases of a chronic HBV infection-immune tolerant and immune active, we used microarray to analyze the whole genome of PBMC that got from hepatitis B virus infected human. We found that FcγRs had a significant difference in different phases of chronic hepatitis B. After that we did PCR to verify the results of whole genome array techniques. The purpose of this paper is to explore FcγRs on different immune cells in different HBV immune status so that we can validate the results of previous experiments and know the value of FcγRs in distinguishing the phases of immune tolerance and immune clearance in hepatitis B.Methods:We used multicolour flowcytometry to detect FcγRⅢ (CD 16) on NK cells and monocytes, FcγRⅡ (CD32) on B cells and monocytes, FcγRI (CD64) on monocytes in different phases of hepatitis B (hepatitis B infection successfully clear group 7patients, hepatitis B clearance group 31 patients, hepatitis B tolerance group 16 patients and healthy controls 20 people). We use Cytometric Bead Array to detect IL-1β, IL-10, MIP-1β, IL-12p70, TNF, IL-6 level in plasma of subjects.The Mann-Whiney U test was used for comparison of continuous data between two groups. The Spearman correlation analysis was used for correlation analysis.Results:1、The patients of immune active group had a lower proportion of CD3-C D56+CD16+ NK cells than patients of immune tolerance (median 6.00% vs 13. 22%,P=0.001)and healthy controls (median 6.00% vs 17.62%, P<0.001). The patie-nts of hepatitis B infection successfully cleared group also had a lower p erc-entage of CD3-CD56+CD16+NK cells than healthy controls (median 5.66% vs 17.62%, P=0.002).2,. The proportion of CD3-CD19+CD32+B cells in immune active group was higer than immune tolerance group (median 11.68% vs 7.28%,.P=0.008). The percentage of CD3-CD5+CD32+B cells in immune active groups was higher than healthy controls (median 3.37% vs 1.46%, P=0.002). The proportion of CD3’CD19+CD5-CD32+B cells was higher in immune active group than immune tolerance group (median 7.77% vs 5.22%, P=0.004).The percentage of CD3CD19+CD5+CD32+B cells was higher in immune active group than healthy controls (median 3.26% vs 1.34%, P=0.006). The proportion of FcyR Ⅱ on CD3CD19+B cells, CD3-CD5+B cells,CD3CD19+D5- B cells,CD3-CD19+D5+ B cells has not significant difference (P>0.0083) when compared any two groups.3、The percentage of CD14highCD16+monocytes was higher in immune clearance group than immune tolerance group (median 8.04% vs 4.84%,P<0.001) and healthy controls (median 8.04% vs 3.59%, P< 0.001). The patients of hepatitis B infection successfully cleared groups also had a higer percentage of CD14highCD16+monocytes than healthy controls (median 7.96% vs 3.59%, P=0.002). When comparing any two groups, there was not significant difference (P>0.0083) in subsets of CD14+CD32+ monocytes and CD14+CD64+ monocytes.4、The proportion of CD14high CD16+monocytes had a positive correlation with ALT level (r=0.694, P< 0.001). The percentage of CD14highCD16+ monocytes had a positive correlation with AST (r=0.698, P<0.001). There was a negative correlation between the subsets of CD14highCD16+ monocytes and serum HBV DNA (r=-0.446, P=0.0017). There was a negative correlation between the subsets of CD 14highCD 16+ monocytes and serum HBsAg (r=-0.614, P<0.001).5、The change of cytokine in four groups. The level of IL-6, IL-10, IL-1β, TNF, MIP-1β and IL-12p70 in immune active group (P<0.001, P<0.001, P<0.001, P=0.001, P< 0.001, P< 0.001) were higher than immune tolerance group and healthy controls (P=0.001, P<0.001, P<0.001, P=0.001, P<0.001, P<0.001)Conclusions:1、The results of this paper verify the conclusion in whole genome array techniques and Real time PCR of FcγRs. The expression of FcγRs is differential in different status of HBV infection.2、The differential expression of FcyRs in different immune cells may be a marker to distinguish HBV immune tolerance and clearance phase. |
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