The Change Of Treg/th17and Tfh Cells In Bone Marrow Failure Diseases

Objective:(1)To examinate the expression of Foxp3and RORγt in CD4⁺T cells inmyelodysplastic syndrome (MDS), aplastic anemia(AA) and immuno-relatedhematocytopenia(IRH) patients and analyze the changes of the subsets of Treg,Th17andTreg/Th17in these diseases.(2)To examinate the expression of Bcl-6in CD4⁺T cells in IRH patients and evaluatethe correlation between T follicular helper cells (Tfh) and IRH.Methods:(1) From January2011to March2012, collect10ml bone arrow, anticoagulated byheparin, of patients with peripheral cytopenia in two or three lineages or iron deficiencyanemia (IDA) patients from the First Affiliated Hospital of Soochow University. Thesamples of IDA patients are served as normal controlgroup.(2) Isolate bone marrow mononuclear cells (BMMNC) by density gradientcentrifugation and seperate CD4⁺T cells with magnetic activated cell sorting (MACS). Thetotal RNA of CD4⁺T cells were purified with trizol and transcripted reversely into cDNA.After that, detect the expression of Foxp3, RORγ and Bcl-6, transcription factor of Treg,Th17and Tfh cells respectively, by the real time fluorescent quantitative-polymerase chainreaction(TagMan)(QRT-PCR).(3) According to the clinical information collected, diagnose these patients as MDS, AAand IRH definitely and respectively.(4) In MDS patients, measure the correlativity between the ratio of bone arrow blast andTreg,Th17and Treg/Th17by linear regression analysis. (5) Compare the average value of Treg/Th17in AA and IRH patients with normalcontrol group by t test.(6) Compare the average value of Bcl-6in IRH patients with normal control group by ttest.Results:(1) In MDS patients, Treg/Th17has a high correlation with the ratio of bone marrowblast (r=0.793).(2) Th17cells in MDS patients with no less than5%bone marrow blast is lower thanthose patients with less than5%bone marrow blast(p=0.007).(3) Treg/Th17in refractory/relapsed AA patients are markedly lower than it in normalcontrol group or newly diagnosed patients(p=0.006ï¼›p<0.001).(4) Compared to normal control group, Tfh cells and Treg/Th17are both significantlyhigher in IRH patients (p=0.016; p=0.045)(5) Th17cells in IRH patients is significantly lower than it in AA group(p<0.001), andTreg/Th17is higher than it in AA group(p=0.002).Conclusion:(1) In MDS patients, the positive correlation between Treg/Th17and the ratio of bonemarrow blast suggest that the inhibition of Th17cells is associated with the progression ofMDS.(2) In refractory/relapsed AA patients, Treg/Th17is unbalanced, which may participatein the hyperactivity of cellular immunity.(3) Unlike AA, the increasing of Tfh cells, not Th17cells, may be the primaryimmunopathogenesis of IRH.