Expression And Significance Of CD4~+CD25~+ Egulatory T Cells In Patients With Primary Immune Thrombocytopenia

BackgroundPrimary immune thrombocytopenia(ITP)is a kind of Immune related hemorrhagic disease,the disease by Platelet autoantibody adjustment,endothelial cells and spleen system Fc receptor related phagocytosis.Autoimmune process is also considered the Pathogenesis of the disease.CD4~+CD25~+Foxp3~+regulator T cells(Treg)play an important role in autoimmune diseases,a growing number of studies have found that patients with ITP in abnormal number of Treg cells,which leads to the disorder of immune tolerance.According to the research of evidence-based medicine,the number and immune function of Treg cells in peripheral blood were changed in patients with ITP after treatment.ObjectivesITP patients,and healthy volunteers,Treg cell expression in peripheral blood were detected by real-time fluorescence quantitative(PCR)technique and flow cytometry(FCM),analysis of Treg cell in ITP patients before and after treatment and the difference between ITP patients and the control group,understand its significance to ITP pathogenesis and corticoid sensitivity.MethodsCase group:according to the inclusion criteria,a total of 40 newly diagnosed ITPpatients were selected from the department of hematology,the first affiliated hospital of xinxiang medical university from January 2016 to August 2017.After 2 weeks of treatment with glucocorticoid,according to the treatment effect,ITP patients were divided into steroid-sensitive group and steroid-insensitive group,among which there were 32 patients in steroid-sensitive group and 8 in insensitive group.Control group:30 healthy volunteers.All patients were informed and signed consent letter.Accurate records of clinical data:age,gender,past medical history and other clinical indicators.Record the clinical indicators:blood routine,T lymphocyte subgroup.real-time PCR was used to detect FOXP3 in peripheral blood before and after the treatment,the FCM was used to detect the ratio of CD4~+CD25~+T cells and CD4~+CD25~+CD127~-Treg cells to CD4~+T cells.Compare the above mentioned inspection indicators in these group,and do statistical analysis.ResoultsCompared with control group,the proportion of CD4~+CD25~+T cells and CD4~+CD25~+CD127~-T cells was lower in the peripheral blood of case group[(14.91±1.96)%vs(12.75±2.42)%,(P=0.015)];[(8.30±1.1)%vs(7.16±2.63)%,(P=0.034)].Compared with steroid-sensitive group,the proportion of CD4~+CD25~+T cells and CD4~+CD25~+CD127~-T cells was lower in the peripheral blood of steroid-insensitive group[(13.80±1.87)%vs(9.86±0.90)%,(P<0.01)];[(7.70±1.7)%vs(5.40±1.02)%,(P<0.01)].Compared with control group,the FOXP3 expression was lower in the peripheral blood of case group[(2.59±1.33)vs(1.49±1.27),(P<0.01)],but increased after treatment[(1.49±1.27)vs(2.12±1.43),(P=0.04)],then no differences between case group after treatment and control were observed[(2.59±1.33)vs(2.12±1.43),(P=0.162)].Before treatment,compared with steroid-sensitive group,the FOXP3 expression was lower in the peripheral blood of steroid-insensitive group[(1.75±1.28)vs(0.45±0.32),(P<0.01)].after treatment,compared with steroid-sensitive group,the FOXP3 expression was lower in the peripheral blood of steroid-insensitive group[(2.48±1.36)vs(0.65±0.29),(P<0.01)].In steroid-insensitive group,the FOXP3 expression has no difference between before and after treatment[(0.65±0.29)vs(0.45±0.32),(P=0.211)];In steroid-sensitive group,compared with before treatment,the FOXP3 expression was higher after treatment[(2.48±1.36)vs(1.75±1.28),(P<0.05)].Compared with control group,the proportion of CD4~+cells was lower in the peripheral blood of case group[(41.8±3.77)%vs(37.82±2.83)%,(P<0.01)],CD8~+cells was higher[(25.9±3.18)%vs(28.84±5.19)%,(P<0.01)],and CD4~+/CD8~+ratio was lower[(1.36±0.27)%vs(1.61±0.59)%,(P<0.01)].Compared with steroid-sensitive group,the proportion of CD4~+cells has no difference with insensitive group in the peripheral blood[(37.96±3.07)%vs(35.98±2.15)%,(P=0.094)],CD8~+cells was higher[(27.71±4.72)%vs(33.35±4.73)%,(P<0.01)],and CD4~+/CD8~+ratio was lower[(1.41±0.27)%vs(1.10±0.16)%,(P<0.01)].Bone marrow megakaryocyte of case group have an impact on the result of the treatment,the patients who bone marrow megakaryocytes were above 35 at admission,were more effective with glucocorticoid than that under or equal of 35,the difference was statistically significant(P<0.05).In the case group,1 case of patients underwent splenectomy,3 patients automatically leaved hospital and 7 cases adopt the second-line treatment,the operation and application of the second line treatment patients are insensitive to glucocorticoid.Conclusions1.CD4+CD25+regulatory T cell significantly decreased in ITP patients.2.CD4+CD25+regulatoryTcellhadasignificantdifferencebetween steroid-sensitive and insensitive patients,and decreased significantly in steroid-insensitive patients,It may seen as an early indicator of patients’corticoid sensitivity.3.The effect of steroid therapy was positively correlated with the number of bone marrow megakaryocytes.