Study on glycolytic metabolism in CD4+T cells in patients with primary immune thrombocytopenia
Objective To investigate the glycolytic metabolic characteristics of CD4+T cells in patients with primary immune thrombocytopenia(ITP)and to elucidate their regulatory role on CD4+T cell function.Methods A total of 92 ITP patients and 46 healthy controls were enrolled in this study.Flow cytometry was used to assess 2-NBDG uptake and the expression levels of glycolytic enzymes(GLUT1,LDHA,PKM2,HK2,and PFKPB3).CD4+T cells were isolated using immunomagnetic beads.Following glucose uptake inhibition in CD4+T cells,flow cytometry method was used to evaluate lymphocyte activation,CCK8 assay for cell proliferation,and ELISA method was used to detect levels of IFN-γ,IL-4,IL-17,and TGF-β1 in the culture supernatant.Results Compared to the healthy control group,ITP patients showed increased glucose uptake in CD4+T cells in vivo,in vitro at rest,and after restimulation(in vivo:9.32%±1.67%vs 3.15%±0.41%,P<0.01;resting:12.92%±2.10%vs 4.87%±1.08%,P=0.02;restimulation:25.52%±3.44%vs 14.71%±3.50%,P=0.03).Expression level of GLUT1 on the cell surface membrane of CD4+T cells was significantly increased in ITP patients(GLUT1:27.70%±2.13%vs 18.12%±2.22%,P<0.01).After inhibiting glucose uptake with 2-DG,the proliferation of CD4+T cells,the proportion of CD25+late-activated T cells were reduced in ITP patients[proliferation(OD):ITP:0.40±0.14 vs 0.25±0.04,P=0.03;activated:ITP:14.65%±2.42%vs 8.71%±1.11%,P<0.01],and the levels of IL-17 in the culture supernatant decreased,while IL-4 levels increased in ITP patients(IL-17:0.70±0.11 ng/L vs 0.44±0.06 ng/L,P=0.03;IL-4:1.36±0.25 ng/L vs 2.20±0.47 ng/L,P=0.02).Conclusion The increased glycolytic metabolic characteristicsin CD4+T cells of ITP patients may contribute to the pathogenesis of ITP by promoting CD4+T cell activation and polarization shift.
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