| Objectives The level of CD4~+CD25~+CD127~-Treg cells and related cytokines in peripheral blood of hemophilia A patients was detected by flow cytometry and enzyme-linked immunosorbent assay(ELISA)to understand the difference of CD4~+CD25~+CD127~-Treg cells and related cytokines in hemophilia A patients with or no inhibitors,and to understand its possible mechanism in the formation of inhibitors.Methods Forty-three hemophilia A patients with complete medical history registered in Tangshan Hemophilia Center(Department of Hematology,affiliated Hospital of North China University of Technology)were selected as subjects,including patients with-inhibitor group(n=6)and patients without inhibitor group(n=37).All the selected patients had informed consent and signed the informed consent form.Record the basic clinical data of the patients:age,height,weight,classification of severity of disease and so on.Flow cytometry was used to detect the expression of CD4~+CD25~+CD127~-Treg cells in peripheral blood of the all patients,and ELISA was adopted to test the expression of serum IL-35,TGF-βand IL-10 in plasma.Finally,the data were analyzed.Results 1 Treg cell expression analysis:the proportion of CD4~+CD25~+CD127~-/CD4~+in hemophilia A patients with-inhibitor group(3.01±0.83)%was significantly lower than that in hemophilia A patients with inhibitor negative group(4.27±0.91)%.The difference was statistically significant(P<0.05).The level of Treg cells decreased and the risk of producing inhibitors increased,and the relationship between them was statistically significant(P<0.05).The proportion of CD4~+CD25~+CD127~-/CD4~+in patients with severe hemophilia A(3.99±1.02)%,that in patients with moderate type was(4.19±0.79)%,and that in patients with mild patients was(4.38±1.01)%.There was no significant difference among the three groups(P>0.05).2 Analysis of expression of related cytokines:the median serum IL-35level was 0.24 ng/ml in hemophilia A patients with inhibitor group,which was lower than that in hemophilia A patients with inhibitor-negative group 0.39 ng/ml.The median level of serum TGF-βand IL-10 in hemophilia A patients with-inhibitor group were 1.16 ng/ml and0.12 ng/ml,which were lower than that in hemophilia A patients without inhibitor group1.55 ng/ml and 0.16 ng/ml,there were statistically significant differences between groups(P<0.05).The levels of IL-35 and TGF-βwere decreased,and the risk of producing inhibitor was increased,and the relationship between them was statistically significant(P<0.05).The expression level of Treg cells was positively correlated with IL-35(r=0.3145,P=0.04),but not with TGF-βand IL-10(r=0.1876,P=0.2283;r=0.1019,P=0.5155).There was no correlation between serum IL-35 expression with TGF-βand IL-10(r=0.0378,P=0.8079;r=-0.0307,P=0.8446),and there was a positive correlation between IL-10 with TGF-β(r=0.9696,P<0.0001).Conclusions 1 The formation of VIII inhibitors in patients with hemophilia A is related to the level of CD4~+CD25~+CD127~-Treg cells in peripheral blood.The level of CD4~+CD25~+CD127~-Treg cells in peripheral blood decreased and risk of producing inhibitors increased.2 The decrease of IL-35 expression in hemophilia A inhibitor-positive patients is related to the formation of inhibitors.3 The level of Treg cells in peripheral blood of patients with hemophilia A is positively correlated with the level of IL-35 expression,but not with the severity of the disease.Figure 3;Table 8;Reference 103... |
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