Dendritic cells phagocytized apoptotic lymphocytes induce the development of regulatory B cells with high secretion of interleukin-10
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摘要:
目的 探讨未成熟树突状细胞(imDC)吞噬经补骨脂素长波紫外线(PUVA)处理的小鼠脾B淋巴细胞(PUVA-SP)后, 将B淋巴细胞诱导为高分泌白细胞介素(IL)-10(IL-10+)的调节性B细胞的可行性。 方法 培养小鼠骨髓来源DC,小鼠脾淋巴细胞经PUVA光源照射得到PUVA-SP,在体外将PUVA-SP与骨髓来源的imDC共同培养,后者吞噬PUVA-SP后形成PUVA-SP DC。应用CD19磁珠分选小鼠脾脏B淋巴细胞,将不同的DC与B淋巴细胞共同培养48 h。采用酶链免疫吸附试验(ELISA)检测B淋巴细胞、imDC、imDC+B淋巴细胞、PUVA-SP DC、PUVA-SP DC+B淋巴细胞的培养上清液中干扰素(IFN)-γ、转化生长因子(TGF)-β、IL-12p70、IL-10等细胞因子的含量。流式细胞仪分析B淋巴细胞、imDC+B淋巴细胞、mDC+B淋巴细胞、PUVA-SP DC+B淋巴细胞共培养后IL-10+调节性B细胞数量。 结果 与其他4组细胞培养液比较,PUVA-SP DC+B淋巴细胞上清液中的IL-10含量明显升高(均为P<0.05)。与其他各组相比,PUVA-SP DC+B淋巴细胞中IL-10+调节性B细胞数量明显增多。 结论 PUVA-SP DC可以诱导脾脏B淋巴细胞分化为IL-10+调节性B细胞。 Abstract:Objective To investigate the feasibility of immature dendritic cells (imDC) phagocytized psoralen ultraviolet A (PUVA)-treated splenic lymphocytes (PUVA-SP DC) in mice inducing B lymphocytes to be regulatory B cells(Breg) with high secretion of interleukin(IL)-10(IL-10+Breg). Methods Bone marrow-derived DC of mice was cultured. Spleen lymphocytes of mice were isolated and treated by PUVA, and turned to be PUVA-SP. The bone marrow-derived imDC was co-cultured with PUVA-SP in vitro to obtain PUVA-SP DC. Splenic B lymphocytes of mice were separated by anti-CD19 magnetic beads and co-cultured with different kinds of DC for 48 hours. The levels of interferon(IFN)-γ, transforming growth factor(TGF)-β, IL-12p70, and IL-10 in the culture supernatant of B lymphocytes, imDC, imDC+B lymphocytes, PUVA-SP DC and PUVA-SP DC+B lymphocytes were measured by enzyme-linked immune absorbent assay(ELISA). The accounts of IL-10+Breg in B lymphocytes, imDC+B lymphocytes, mDC+B lymphocytes and PUVA-SP DC+B lymphocytes were detected by flow cytometry. Results Compared with the other 4 groups, the level of IL-10 in cell culture supernatant of PUVA-SP DC+B lymphocytes was significantly higher(all in P<0.05). Compared with the other groups, the account of IL-10+Breg in PUVA-SP DC+B lymphocytes was significantly higher. Conclusions PUVA-SP DC can induce splenic B lymphocytes to differentiate into IL-10+Breg. -
Key words:
- Dendritic cell /
- Photochemical method in vitro /
- Regulatory B cell /
- Interleukin-10
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