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目的初步探讨人T淋巴细胞活化后细胞内5-羟甲基胞嘧啶(5-hydroxymethylcytosine,5-hmC)和5-甲基胞嘧啶(5-methylcytosine,5-mC)的含量变化。方法成年健康人外周血单个核细胞(peripheral blood mononuclear cell,PBMCs)分别经刀豆蛋白(ConA)或CD3抗体和CD28抗体处理72 h后,采用免疫荧光法观察5-hmC和5-mC的变化,Real-time PCR检测TETs的mRNA转录水平。结果免疫荧光结果显示无论是经ConA非特异性刺激还是经CD3抗体和CD28抗体处理的人T淋巴细胞细胞核内,5-hmC水平明显高于5-mC,且后者从核内转位到胞浆;CD3抗体和CD28抗体处理后的TET2 mRNA转录水平显著高于非处理组。结论活化的T淋巴细胞中5-mC转变是由TET2调控的。
Objective To investigate the changes of intracellular levels of 5-hydroxymethylcytosine (5-hmC) and 5-methylcytosine (5-mC) after human T lymphocyte activation. Methods Peripheral blood mononuclear cells (PBMCs) from healthy adults were treated with ConA or CD3 antibody and CD28 antibody for 72 hours respectively. The changes of 5-hmC and 5-mC were observed by immunofluorescence staining Real-time PCR detection of TETs mRNA transcription level. Results Immunofluorescence results showed that the 5-hmC level was significantly higher than that of 5-mC in the nuclei of human T lymphocytes stimulated by non-specific stimulation with ConA or by CD3 and CD28 antibodies, and the latter translocated from the nucleus to the cytoplasm ; CD3 antibody and CD28 antibody TET2 mRNA transcription levels were significantly higher than the untreated group. Conclusion The 5-mC transition in activated T lymphocytes is regulated by TET2.