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目的:研究可溶性Jagged 1/Fc嵌合蛋白在小鼠淋巴细胞活化、增殖和周期中的作用。方法:选择有效剂量为500μg/L Jagged 1/Fc嵌合蛋白(Jagged 1/Fc),以活体染料/羧基荧光素乙酰乙酸琥珀酰亚胺酯染色,建立在多克隆刺激剂刀豆蛋白A(ConA)或佛波醇酯(PDB)加离子霉素(Ion)刺激下评价淋巴细胞增殖的模型,通过流式细胞术分析Jagged 1/Fc对淋巴细胞增殖的作用;采用碘化丙锭染色检测Jagged 1/Fc对PDB加Ion刺激的淋巴细胞周期变化的影响;利用荧光标记的mAb双染技术观察Jagged 1/Fc对T细胞早期和中期活化标志CD69和CD25表达的影响。结果:Jagged1/Fc对ConA刺激或非刺激的CD3+细胞CD69和CD25的表达无明显影响,对ConA或PDB加Ion刺激或非刺激的淋巴细胞的增殖指数也无明显影响,导致非刺激的淋巴细胞sub-G0期细胞百分比增高,S期细胞百分比降低,但并不影响PDB加Ion诱导的淋巴细胞各期的变化。结论:这些结果提示500μg/L Jagged 1/Fc对小鼠淋巴细胞活化和增殖无明显作用,对PDB加Ion刺激的淋巴细胞周期也无明显影响,但能促进未受刺激的淋巴细胞凋亡,使细胞停滞于G0/G1期,阻止其进入S期。
AIM: To investigate the role of soluble Jagged 1 / Fc chimeric protein in the activation, proliferation and cycle of mouse lymphocytes. METHODS: Jagged 1 / Fc (500μg / L) was selected and stained with live dye / carboxyfluorescein acetoacetate succinimidyl ester. The polyclonal agonist concanavalin A ConA or PDB plus ionomycin (Ion) to evaluate the proliferation of lymphocytes. The effect of Jagged 1 / Fc on lymphocyte proliferation was analyzed by flow cytometry. Propidium iodide staining The effect of Jagged 1 / Fc on the PDB plus Ion-stimulated lymphocyte cycle was investigated. The effect of Jagged 1 / Fc on the expression of CD69 and CD25 on early and mid-term T cells was observed by fluorescence-labeled mAb double staining. Results: Jagged1 / Fc had no significant effect on the expression of CD69 and CD25 on ConA-stimulated or non-stimulated CD3 + cells and had no significant effect on the proliferation index of ConA or PDB plus Ion-stimulated or non-stimulated lymphocytes, resulting in non-stimulated lymphocytes The percentage of cells in sub-G0 phase increased and the percentage of S phase cells decreased, but did not affect the changes of lymphocytes induced by PDB plus Ion. CONCLUSION: These results suggest that 500μg / L Jagged 1 / Fc has no effect on lymphocyte activation and proliferation in mice, but also has no significant effect on PDB plus Ion-stimulated lymphocyte cycle, but can promote unstimulated lymphocyte apoptosis, The cells are arrested in G0 / G1 phase, preventing them from entering the S phase.