The Role Of Tfh Cell And B Cell In The Progression Of Chronic Graft Versus Host Disease In Human

Part I The role of Tfh cell and B cell in the pathophysiology of chronic graft versus host disease in humanObjective:To explore the roles and mechanisms of circulating follicular helper T(cTfh)cells,extrafollicular helper T cells and B cells in human chronic graft-versus-host disease(cGVHD).Methods:We collected the peripheral blood of patients undergoing allogeneic hematopoietic stem cell transplantation(allo-HSCT)in our hospital from 2016 to 2019.Sorting mononuclear cells(MNCs)using ficoll density gradient centrifugation,then detected the percentage of cTfh cells,extrafollicular helper T cells and B cells in patients with cGVHD using flow cytometry.In addition,peripheral blood of healthy donors was identically detected as control.The unpaired student T test and the Mann-Whitney U test in SPSS version 20.0 were used to compare the two groups.Spearman correlation analysis was also used to analyze the correlation of cells.Results:In our study,there were 64 patients with active cGVHD enrolling,of which 37 were male and 27 were female,with a median age of 37.5(10-64)years,median specimen collection was 11.87(3.50-33.67)months away from transplantation.There were 41(64.1%)cGVHD patients involved one organ,16(25%)involved two organs,and 7(10.9%)involved three or more organs.cTfh cells and extrafollicular helper T cells were detected in a total of 64 patients,of which 20 were detecting B cell subsets.Our results show:1.cTfh(CD4+CXCR5+)cells,ICOS+cTfh cells and Ki-67+cTfh cells decreased significantly in cGVHD patients(P<0.05).The expression of PD1+cTfh cells decreased in cGVHD patients,the difference between the two groups was not statistically significant(P>0.05);but the expression of IL-21+cTfh cells was higher than that of the control group,and the difference was not statistically significant(P>0.05).Correlation analysis found expression of Ki-67+cTfh cells and PD1+cTfh cells were positively correlated in cGVHD group(r=0.4896,P<0.05).2.The CD44+CD62L-PSGL-1-CD4+T cells were detected in both cGVHD patients and healthy donors,the expression of the cells was significantly increased in cGVHD patients compared with healthy donors(P<0.05).3.There were B subsets changing in peripheral blood of cGVHD patients,and two B cells with CD27 negative expression were decreased in cGVHD patients.The expression of naive B cells(CD19+IgD+CD38lo CD27-)was reduced significantly(P<0.05),but tra-nsitional B cells(CD19+IgD+CD38hi CD27-)were no significant difference between the two groups(P>0.05).However the expression of two CD27 positive B cells were signific-antly higher than those of the control group(P<0.05),and the two kinds of cells were pre-GC B cells(CD19+IgD+CD38hi CD 27+)and plasmablasts/plasma cells(PB/PC,CD19+IgDlo/-CD38hi CD27+).4.We analyzed the correlation between cTfh cells proliferation and activation subtypes and extrafollicular helper T cells with the percentage of PB/PC.The expression of extrafollicular helper T cells and Ki-67+cTfh cells were positively correlated with PB/PC(r=0.5412,P<0.05;r=0.6486,P<0.05,respectively).Conclusion:In cGVHD patients,the expression of cTfh cells decreased,and the expression of extrafollicular helper T cells increased,interacting with B cells and participating in the development of cGVHD.Part ? Expression of lymphocyte subsets in chronic graft-versus-host disease after transplantationObjective: To investigate the expression level of lymphocyte subsets in peripheral blood of patients with cGVHD after transplantation and to analyze the correlation of lymphocyte subsets and cGVHD.Methods:The clinical data and laboratory data of allo-HSCT recipients who underw?ent post-transplantation imnmne ftmction examination in the blood stream cell laboratory of our hospital from March 2019 to August 2019 were collected.The patients were divided into cGVHD group and no cGVHD group,and compared the difference of lymphocyte subsets between the two groups.The unpaired student T test and Mann-Whitney f/test of the SPSS version 20.0 were used in group comparison.Results: In the study,there were 53 post-transplantation patients from January 2013 to November 2018 enrolling,and the period of post-transplantation immune function test from March 2019 to August 2019,the median time of immune flmction examination to transplantation was 20.7(6.7-77)months.The median age was 29(16-50)years.Of the 53 patients,26 were cGVHD patients,of which 15(57.7%)were involved in one organ,3(11.5%)were involved in 2 organs,and 8(30.8%)were involved in 3 or more organs;There were 4(15.4%)cases at 6-12 months,13(50%)cases at 1-2 years,and 9(34.6%)cases more than 2 years after transplantation.There were 27 patients without cGVHD,of which 3(11.2%)were 6-12 months after transplantation,12(44.4%)were 1-2 years,and12(44.4%)were more than 2 years after transplantation.The results show that:1.The percentage and cell count of CD4+ T lymphocytes in the cGVHD group were significantly lower than those in the non-cGVHD group,and the differences were statistically significant(P<0.05);the percentage and cell count of CD8+ T lymphocytes were not statistically different between the two groups(P>0.05);The percentage of regulatory T cells(CD3+ CD4+ CD25+ CD 127=)showed a decreasing trend in the cGVHD group,the difference was not statistically significant(P>0.05),and its cell count was significantly reduced in the cGVHD group,and the difference between the two groups was statistically significant(P<0.05).2.The percentage and cell count of B lymphocytes in the cGVHD group were significantly lower than those in the non-cGVHD group,and the differences were statistically significant(P<0.05).The percentage of naive B lymphocytes(CD19+ CD27"IgD+)showed a decreasing trend in the cGVHD group,the difference was not statistically significant(P>0.05),the cell count was decreased in the cGVHD group,and the difference was statistically significant(P<0.05);the percentage of immature B lymphocytes(transitional,CD19+ CD27' CD38 hlgh CD24+)was in the decreasing trend in the cGVHD group,there was no statistically significant difference(P>0.05).The cell count decreased in the cGVHD group,and the difference was statistically significant(P<0.05).The percentage of MZ B lymphocytes(CD19+ CD27+ IgD+)in the cGVHD group showed an upward trend,the difference was not statistically significant(P>0.05);The percentage of classic swiched B lymphocytes(CD19+ 0027^ CD38 dun IgD ")in the cGVHD group showed an increasing trend,the difference was not statistically significant(P>0.05);The percentage and cell count of plasmablasts(CD19+ CD27 Mgh CD38hlgh IgD")showed an increasing trend in the cGVHD group,and the differences between the two groups were not statistically significant(P>0.05).Conclusion: The proportion of lymphocyte subsets in cGVHD and non-cGVHD patients after allo-HSCT follow-up in this study.Compared with non-cGVHD patients,the proportion and cell count of cGVHD patients with regxilatory T cells,naive B lymphocytes and immature lymphocytes decreased,and the proportion of MZ B lymphocytes and classic switched lymphocytes increased,and the changes of lymphocyte subsets after transplantation can be dynamically monitored clinically to judge the immune fimction status of patients.