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目的探讨 Notch 配体δ-1在红系造血细胞分化过程中对可溶性 IL-6受体(sIL-6R)生物效应的影响。方法用 CD34免疫磁珠和 FACS Vantage 流式细胞仪筛选脐血单个核细胞中的CD34~+CD38~-细胞;将 CD34~+CD38~-细胞用含 SCF、F1t3L、TPO 和 IL-3四种生长因子组合(4GFs)的培养基培养7 d,然后用 CD36免疫磁珠分离 CD36~+红系祖细胞,用流式细胞术检测 IL-6R 和血型糖蛋白(GPA)的表达,并分选 CD36~+GPA~-IL-6R~+和 CD36~+GPA~-IL-6R~-细胞,将这两种表型的细胞分别进行集落分析;将 CD36~+GPA~-IL-6R~-细胞用含有4GFs、4GFs+IL-6或4GFs+IL-6/sIL-6R 融合蛋白(FP6)培养基并在加与不加 Notch 配体δ-1的情况下培养14 d,并对 CD36~+GPA~(high)成熟红细胞进行计数。结果 CD36~+GPA~+细胞中 IL-6R~-细胞占95&;CD36~+GPA~-细胞可分为 IL-6R~+和 IL-6R~-两部分,分别占46%和54%;IL-6R~+细胞形成的粒-单核细胞集落形成单位(CFU-GM)数为2.1±1.8,IL-6R~-细胞形成的红系祖细胞爆式集落形成单位(BFU-E)数为58.2±18.1,明显高于 IL-6R~+细胞形成的 CFU-GM 数(P<0.05);在含有 FP6的培养体系中,CD36~+GPA~(high)细胞计数为(1.400±0.180)×10~6;在含 FP6和 Notch 配体δ-1的培养体系中,CD36~+GPA~(high)细胞计数为(2.460±0.190)×10~6,明显高于单独含有 FP6的培养体系(P<0.05)。结论 Notch 配体δ-1可增强 IL-6~-红系细胞分化过程中sIL-6R 所介导的生物学效应。
Objective To investigate the effect of Notch ligand δ-1 on the biological effects of soluble IL-6 receptor (sIL-6R) during erythroid hematopoietic differentiation. Methods CD34 ~ + CD38 ~ - cells in umbilical cord blood mononuclear cells were screened by CD34 immunomagnetic beads and FACS Vantage flow cytometry. Four types of CD34 ~ + CD38 ~ - cells were stained with SCF, F1t3L, TPO and IL- (4GFs) for 7 days. CD36 ~ + erythroid progenitor cells were isolated by CD36 immunomagnetic beads. The expression of IL-6R and GPA was detected by flow cytometry and sorted CD36 ~ + GPA ~ -IL-6R ~ + and CD36 ~ + GPA ~ -IL-6R ~ - cells were used for colony analysis. Cells were cultured for 14 days in medium containing 4GFs, 4GFs + IL-6 or 4GFs + IL-6 / sIL-6R fusion protein (FP6) and with or without Notch ligand δ- + GPA ~ (high) mature red blood cells were counted. Results CD36 ~ + GPA ~ - cells could be divided into IL-6R ~ + and IL-6R ~ -, accounting for 46% and 54%, respectively. The number of granulocyte-mononuclear cell colony forming units (CFU-GM) formed by IL-6R ~ + cells was 2.1 ± 1.8. The number of erythroid progenitor blast-forming units (BFU-E) (58.2 ± 18.1), which was significantly higher than that of IL-6R + cells (P <0.05). In the culture system containing FP6, the number of CD36 + GPA high cells was (1.400 ± 0.180) × 10 ~ 6. In the culture system containing FP6 and Notch ligand δ-1, the number of CD36 ~ + GPA ~ (high) cells was (2.460 ± 0.190) × 10 ~ 6, which was significantly higher than that of FP6 (P <0.05). Conclusion Notch ligand δ-1 can enhance the biological effects mediated by sIL-6R during the differentiation of IL-6-erythroid cells.