| Objective To explore the level of transcription factor Bach2, Blimp-1 and hemeoxygenase-1(HO-1) in peripheral blood(PB) CD4~+ or CD8~+ T lymphocytes from patients with severe aplastic anemia(SAA) and in CD4~+ or CD8~+ T lymphocytes and CD19~+ B lymphocytes from patients with immune-related pancytopenia syndrome(IRP).Methods 10 untreated SAA and 10 untreated IRP patients, 10 remission patients, as well as 10 normal controls of Tianjin Medical University General Hospital from August 2014 to March 2016 were entrolled in this study. Part â… The CD4~+ or CD8~+T cells from 10 SAA patients, 10 remission SAA patients and 10 normal controls were sorted by magnetic activated cell sorting system(MACS), and the m RNA expression of Bach2 were analyzed by Quantitative real-time PCR. Function molecules(perforin and granzyme B) of peripheral blood CD8~+ T cells from SAA patients, remission group and normal controls were detected by Quantitative real-time PCR. Bach2 of peripheral blood CD4~+ or CD8~+ T cells were detected by flow cytometry. Correlation of Bach2 in CD8~+ T cells and function molecules of CD8~+ T cells was analyzed. Partâ…¡ The CD4~+ or CD8~+T cells from 10 IRP patients, 10 remission IRP patients and 10 normal controls were sorted by magnetic activated cell sorting system(MACS), and the m RNA expression of Bach2 were analyzed by Quantitative real-time PCR. Bach2 of peripheral blood CD4~+ or CD8~+ T cells and CD19~+ B cells were detected by flow cytometry. Partâ…¢ The CD4~+ or CD8~+T cells from 10 SAA/IRP patients, 10 remission SAA/IRP patients and 10 normal controls were sorted by magnetic activated cell sorting system(MACS), and the m RNA expression of Blimp-1 and HO-1 were analyzed by Quantitative real-time PCR. Blimp-1 and HO-1 of peripheral blood CD4~+ or CD8~+ T cells from all patients and CD19~+ B cells from IRP patients were detected by flow cytometry. Correlation of Bach2 and Blimp-1/HO-1 in T cells and B cells were analyzed.Results Part â… (1) The relative Bach2 m RNA levels was evidently decreased in the peripheral blood CD4~+ or CD8~+T cells from untreated cases compared to normal controls(P<0.05). After immune suppressive therapy, the expression of Bach2 increased.(2)The median expressions of Bach2, in CD4~+T cells and in CD8~+T cells were 39.04%and 40.86% in newly diagnosed SAA patients, and 53.64% and 64.98% in normal controls. The expression of Bach2 was lower in newly diagnosed cases than that in healthy controls(P<0.05).(3) We found a negative correlation between the expression of perforin in CD8~+ T cells and the expression of Bach2 in CD8~+ T cells in SAA patients and controls(r=-0.684, P<0.05). Meanwhile, the expression of Bach2 in CD8~+ T cells is also negatively associated with expression of granzyme B in CD8~+ T cells(r=-0.389, P<0.05). Part â…¡(1) There was no significantly difference between normal group and untreated IRP group of the relative Bach2 m RNA levels in the peripheral blood CD4~+ or CD8~+T cells and CD19~+ B cells(Pï¹¥0.05).(2)The median expressions of Bach2, in CD4~+T cells and in CD8~+T cells were 44.53%and 59.23% in newly diagnosed IRP patients, and 53.64% and 64.98% in normal controls, there was no significantly difference between normal group and untreated IRP group(P>0.05). The median expressions of Bach2, in CD19~+ B cells was 86.57% in newly diagnosed IRP patients, and 85.14% in normal controls, there was no significantly difference between normal group and untreated IRP group(P>0.05).(3) The relative Bach2 m RNA levels expression of Bach2 was lower in newly diagnosed SAA cases than that in newly diagnosed IRP(P<0.05). Part â…¢(1) The QPCR detected m RNA expression level and the flow cytometry result of Blimp-1 and HO-1 in peripheral blood CD4~+ and CD8~+ T lymphocytes of SAA patients were significantly higher than normal controls(P<0.05), and significantly decreased after immunity treatment(P<0.05).(2)The QPCR detected m RNA expression level and the flow cytometry result of Blimp-1 in peripheral blood CD4~+ and CD8~+ T lymphocytes of IRP patients were not significantly different than normal controls(P>0.05). The flow cytometry result of Blimp-1 and HO-1 in peripheral blood CD19~+ B lymphocytes of IRP patients were significantly higher than normal controls(P<0.05), and significantly decreased after immunity treatment(P<0.05). The QPCR detected m RNA expression level and the flow cytometry result of HO-1 in peripheral blood CD4~+ T lymphocytes of IRP patients were significantly higher than normal controls(P<0.05), and significantly decreased after immunity treatment(P<0.05). The QPCR detected m RNA expression level and the flow cytometry result of HO-1 in peripheral blood CD8~+ T lymphocytes of IRP patients were not significantly different than normal controls(P>0.05).(3) The Blimp-1 and HO-1 in CD4~+ and CD8~+ T lymphocytes of all the observed patients were significantly negative correlation with Bach2(P<0.05). The Blimp-1 in CD19~+ B lymphocyte of IRP patients was significantly negative correlation with Bach2(P<0.05). HO-1 and Bach2 was not significant negative correlation(P>0.05).Conclusions(1) The QPCR detected m RNA expression level and the flow cytometry result of Bach2 in peripheral blood CD4~+ and CD8~+ T lymphocytes of newly diagnosed SAA patients were significantly lower than normal controls, and significantly increased after immunity treatment.(2) The QPCR detected m RNA expression level and the flow cytometry result of Bach2 in peripheral blood CD4~+,CD8~+ T lymphocytes and CD19~+ B lymphocytes of IRP patients were no significant difference compared with normal controls. The QPCR detected m RNA expression level and the flow cytometry result of Bach2 in peripheral blood CD8~+ T lymphocytes of newly diagnosed SAA patients were significantly lower than newly diagnosed IRP patients.(3) The QPCR detected m RNA expression level and the flow cytometry result of Blimp-1 and HO-1 in peripheral blood CD4~+ and CD8~+ T lymphocytes of SAA patients were significantly higher than normal controls, and significantly decreased after immunity treatment.(4) The QPCR detected m RNA expression level and the flow cytometry result of Blimp-1 in peripheral blood CD4~+ T and CD8~+ T lymphocytes of IRP patients were no significant difference compared with normal controls. The flow cytometry result of Blimp-1 in peripheral blood CD19~+ B lymphocytes of IRP patients were significantly higher than normal controls, and significantly decreased after immunity treatment. The flow cytometry result of HO-1 in peripheral blood CD4~+ T and CD19~+ B lymphocytes of IRP patients were significantly higher than normal controls, and significantly decreased after immunity treatment. The QPCR detected m RNA expression level and the flow cytometry result of HO-1 in peripheral blood CD8~+ T lymphocytes of IRP patients were no significant difference compared with normal controls.(5) The Blimp-1 and HO-1 in CD4~+ and CD8~+ T lymphocytes of all the observed patients were significantly negative correlation with Bach2. The Blimp-1 in CD19~+ B lymphocyte of IRP patients was significantly negative correlation with Bach2. HO-1 and Bach2 was not significant negative correlation. |
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