| Objective: Liver transplantation is the only effective method for the treatment of end-stage liver disease.After liver transplantation,especially for patients with liver failure caused by hepatitis B virus(HBV),the biggest problem is HBV recurrence.Hepatitis B vaccination after liver transplantation is an excellent way to prevent HBV recurrence.Hepatitis B vaccine is available for liver transplant recipients,and a certain proportion of patients will produce hepatitis B surface antibody,and the recipients will have active immunity to HBV.Approximately 60%-70% of liver transplantation patients after taking standard hepatitis B vaccinations lack or do not produce insufficient antibody,which is much higher than the figure of healthy vaccine takers at the rate of 5%-10%.However,the mechanism and principle are still unknown.The prognosis of patients with HBV recurrence after liver transplantation is poor,which could even threaten the lives of patients.To explore the mechanism of HBV recurrence after liver transplantation and carry out immunological prevention and treatment is an urgent problem to be solved clinically.Follicular helper T cell(Tfh)is a newly discovered CD4+ T cell subpopulation in recent years,which plays a role of auxiliary B cells in immune response.The process of antibody response is very important and is closely related to the occurrence and development of many diseases.Based on the above metioned reasons,the study of immune responses induced by hepatitis B vaccination after liver transplantation can promote the understanding of the issues.The antibody production will be detected after hepatitis B vaccination in liver transplant recipients,to reveal whether they are affected by Tfh cell subpopulation,to clarify the expression of Tfh cell subpopulation in the related immune response and the influence of peripheral blood Tfh cell cytokines on B cells,which can provide a theoretical basis for long-term survival rate after liver transplantationMethods:1.The subjects of this study are selected patients who have had liver transplantation in the Affiliated Hospital of Qingdao University from 2016 to 2019.All participants met the following criteria:(1)history of liver transplantation;(2)diagnosed with hepatitis B before transplantation(3)Hepatitis B vaccination after liver transplantation;(4)2-3 years after having the vaccination.The basic information of age,gender and history of hepatitis B vaccination was obtained through questionnaires.Participants were divided into response group(>100 IU / L)and non-response group(< 10 IU / L)according to antibody titer,with an average age of 51 years(41-75 years)and gender of 7 / 2(male / female).2.The Tfh cell subpopulation were categorized according to the frequency of Tfh cell subpopulation and B cells detected by flow cytometry.The Tfh cell subpopulation(CD3+CD4+CXCR5+)were identified by the expression of CD3,CD4,CXCR3,CXCR5 and CCR6.Tfh1(CD3+CD4+CXCR3+CXCR5+CCR6-),Tfh2(CD3+CD4+CXCR3-CXCR5+CCR6-),Tfh17(CD3+CD4+CXCR3-CXCR5+CCR6+)and B cells(CD19+)were labeled by CD19.3.Hepatitis B antibody and related immune factors were detected by ELISA.Hepatitis B surface antibody,IL-21 and IL-4 ELISA kit were used to detect hepatitis B surface antibody,IL-21 and IL-4 in the peripheral blood of subjects.4.Correlation analysis between Tfh cell subpopulation and hepatitis B surface antibody production.The correlation between Tfh cell subpopulation and hepatitis B surface antibody titers in subjects was analyzed.The correlation between cytokines IL-21 and IL-4 secreted by Tfh in peripheral blood and the cell frequency of B cells was analyzed.Results: 1.CD3+CD4+T cells were correlated with hepatitis B vaccine response.The frequency of CD3+CD4+T cells in hepatitis B vaccine recipients after liver transplantation was significantly higher than that in the non-responders 2.Tfh were correlated with hepatitis B vaccine response.The frequency of Tfh in the hepatitis B vaccine responder group after liver transplantation was significantly higher than that in the non-responders group,and there were significant differences in the cell frequency of Tfh in the lymphocytes and the cell frequency in CD4+T cells between the two groups(t=2.534,P<0.05).3.The role of Tfh cell subpopulation in hepatitis B vaccine response.There was no significant difference in the frequency of Tfh2 in Tfh between the two groups after liver transplantation(t=0.853,P>0.05).The cell frequency of Tfh1 in Tfh was different between the two groups,and the frequency of Tfh1 cells in hepatitis B vaccine responder group after liver transplantation was significantly higher than that in the non-responders group(t=3.038,P<0.05).The cell frequency of Tfh17 in Tfh was different between the two groups,and the frequency of Tfh17 in the hepatitis B vaccine responder group after liver transplantation was significantly lower than that in the non-responders group(t=3.872,P<0.05).4.Tfh1/Tfh17 was correlated with hepatitis B vaccine response.In the response group of hepatitis B vaccination recipients after liver transplantation,the titer of antibody was positively correlated with Tfh1/Tfh17,and increased as the Tfh1/Tfh17 increased(r=0.9824,P<0.05).There was no significant correlation with the frequency of Tfh2 cells.5.The role of IL-21 and IL-4 in hepatitis B vaccine response.The concentration of IL-21 in peripheral blood was different between the response group and the non-response group,and the concentration of IL-21 in the response group was significantly higher than that in the non-response group(P<0.05).Similarly,the concentration of IL-4 in peripheral blood was different between the response group and the non-response group.The concentration of IL-4 in the response group was significantly higher than that in the nonresponse group(P< 0.01)6.B cells were correlated with cytokines of Tfh in peripheral blood.In the hepatitis B vaccine recipients after liver transplantation,the frequency of B cells in peripheral blood was positively correlated with the concentration of Tfh cytokine IL-21 and IL-4 in peripheral blood,and increased with the increase of both.Conclusion:.1.The frequency of Tfh in the responder group after hepatitis B vaccinationwas significantly higher than that in the non-responders’ group,and there were significant differences in the cell frequency of Tfh in the lymphocytes and the cell frequency in CD4+T cells between the two groups.It was suggested that the production of hepatitis B surface antibody in hepatitis B vaccine recipients after liver transplantation may depend on Tfh,providing clues for the pathogenesis of HBV recurrence after liver transplantation.2.The occurrence of hepatitis B surface antibody in hepatitis B vaccine recipients after liver transplantation depended on the cell frequency of Tfh1 in peripheral blood,and the shift of Tfh1 to Tfh17 ratio may also affect the production of hepatitis B surface antibody.It provides a new theoretical basis for further revealing the local immune microenvironment of liver after liver transplantation.3.Cytokines IL-21 and IL-4 secreted by Tfh in peripheral blood were closely related to the proliferation of B cells and promoted the proliferation of B cells.It provides clues for clinical treatment... |
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