The Study On The Number And Function Of Th9 Cells In The Patients With Immune-related Pancytopenia

Objective:The study is to explore the role of Th9 cells in the pathogenesis of IRP and its mechanism,so as to provide theoretical basis for the diagnosis and treatment of IRP.The number of helper T lymphocyte 9(Th9)and its cytokine interleukin-9(IL-9)in peripheral blood of patients with immune-related pancytopenia(IRP)were detected.The expression of differentiation-related transcription factors PU.1 and BATF in Th9cells was detected by RT-PCR.Methods:Fifty patients with IRP from July 2017 to January 2018 in Tianjin Medical University General Hospital,thirty were newly diagnosed as the newly diagnosed treatment group,twenty were the remission group,and twenty health were enrolled in the control group.The percentage of Th9 cells(CD3~+CD4~+IL-9~+)in peripheral blood of each group was detected by flow cytometry and the level of serum IL-9 of different groups was detected by ELISA.RT-PCR was used to detect the expression of differentiation-related transcription factors PU.1 and BATF in CD4~+cells from different groups.Results:1.The percentage of Th9 cells in peripheral blood of IRP patients in all CD4~+T lymphocytes.The percentage of Th9 cells in the newly diagnosed group was(2.73±1.96)%,the remission group was(1.21±0.86)%,and the control group was(0.68±0.40)%.The percentage of Th9 cells in the newly diagnosed group was significantly higher than that in the remission group(p<0.01),the remission group was significantly higher than that in the control group(p<0.05),and the newly diagnosed group was significantly higher than that in the control group(p<0.001).2.ELISA was used to detect the level of IL-9 in peripheral blood serum of patients with IRP.The level of IL-9 in the newly diagnosed group was(183.91±112.42)pg/ml,the level of IL-9 in the remission group was(105.96±64.79)pg/ml,and the control group was(56.03±14.49)pg/ml;the level of IL-9 in the newly diagnosed group was significantly higher than that in the remission group(p<0.01);the level of IL-9 in the newly diagnosed group was significantly higher than that in the control group(p<0.001).The remission group was also significantly higher than that in the control group(p<0.01).3.PCR was used to detect the relative expression of PU.1 and BATF in peripheral blood CD4+cells.The relative expression of PU.1 in the newly diagnosed group was117.30±82.33,the remission group was 45.46±33.22,and the control group was19.07±13.56.The relative expression of PU.1 in the newly diagnosed group was significantly higher than that in the remission group(p<0.001),the newly diagnosed group was significantly higher than the control group(p<0.01),and the remission group was significantly higher than that in control group(p<0.01);the relative expression of BATF in the newly diagnosed group was 608.57±517.68,the remission group was 238.89±206.80,and the control group was 134.71±113.31.The newly diagnosed group was significantly higher than in the remission group(p<0.01).There was no significant difference in the relative expression of BATF between the remission group and the control group(p>0.05).4.The correlation analysis between serum level of IL-9 and clinical related characters showed that the level of IL-9 was negatively correlated with hemoglobin(p<0.01,r=-0.457),red blood cell count(p<0.05,r=-0.467),platelet count(p<0.05,r=-0.297)and white blood cell count(p<0.05,r=-0.227);Serum interleukin-9 concentration was positively correlated with the percentage of CD5+CD19+/CD19+cells(p<0.05,r=0.376)and the percentage of CD19~+cells(p<0.05,r=0.318).5.The percentage of Th9 cells in peripheral blood was negatively correlated with hemoglobin,(p<0.05,r=-0.366),red blood cell count(p<0.05,r=-0.364),platelet count(p<0.05,r=-0.457)and white blood cell count(p<0.05,r=-0.418).The percentage of Th9 cells was positively correlated with the percentage of CD5~+CD19~+/CD19~+cells(p<0.05,r=0.377)and the percentage of CD19~+cells(p<0.001,r=0.835).Conclusions:1.The percentage of Th9 cells in peripheral blood in CD3~+CD4~+cells in the patients with IRP was significantly higher than the control group.2.The concentration of IL-9 in the patients with IRP was significantly higher than the control group.3.The expression of transcription factors PU.1 and BATF related to Th9 cell differentiation in the IRP patients was significantly higher than the control group.4.The serum levels of IL-9 were negatively correlated with Hb,WBC,RBC and PLT,and positively correlated with the percentage of CD19~+cells and the percentage of CD5~+CD19~+/CD19~+cells.5.The percentage of Th9 cells in IRP patients was negatively correlated with Hb,WBC,RBC and PLT,and positively correlated with the percentage of CD19~+cells and the percentage of CD5~+CD19~+/CD19~+cells.