首页> 美国卫生研究院文献>Analytical Cellular Pathology : the Journal of the European Society for Analytical Cellular Pathology >Comparative Intracellular Cytokine Production by In Vitro Stimulated T Lymphocytes from Human Umbilical Cord Blood (HUCB) and Adult Peripheral Blood (APB)
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Comparative Intracellular Cytokine Production by In Vitro Stimulated T Lymphocytes from Human Umbilical Cord Blood (HUCB) and Adult Peripheral Blood (APB)

机译:从人脐带血(HUCB)和成人外周血(APB)的体外刺激T淋巴细胞比较细胞内细胞因子的生产

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摘要

To date over 400 HUCB transplants have been reported from different centers. It has been suggested that there is a reduced graft‐versus‐host‐disease (GVHD) with HUCB compared to bone marrow transplantation. Since cytokine production by a cell is an indication of the cells function it is important to determinate the differences between APB and HUCB with respect to production of these soluble factors. Our aim was to analyse the intracellular cytokine production by HUCB and APB T lymphocytes with and emphasize on their possible role in GVHD.Heparinized HUCB samples from 8 normal full‐term deliveries and 10 normal blood donors were stimulated 4 hours at 37∘C and 5% CO2 with phorbol 12‐myristate 13‐acetate (PMA) and Ionomycin in the presence of brefeldine. Afterwards cells were stained with CD3, CD4 or CD8 in different combinations. Finally, after cell permeabilization, cells were stained with Il‐2, Il‐4 or IFN‐γ. Data acquisition was performed on a FACScan flow cytometer.Compared to APB, HUCB T lymphocytes produced less Il‐2, Il‐4 and IFN‐γ. In HUCB, Il‐2, Il‐4 and IFN‐γ were produced predominantly by CD4+ T cells. In APB, Il‐2 and Il‐4 were also produced predominantly by CD4+ cells compared with CD8+ T lymphocytes, however, IFN‐γ was produced by both CD4+ and CD8+ T cells. These results indicate that there are clear differences in the cytokine profile between T cells in APB and HUCB.
机译:迄今为止,已从不同中心报告了400多种HUCB移植。与骨髓移植相比,HUCB的移植物抗宿主病(GVHD)减少。由于细胞产生的细胞因子是细胞功能的指标,因此重要的是确定APB和HUCB在产生这些可溶性因子方面的差异。我们的目的是分析HUCB和APB T淋巴细胞在GVHD中的细胞内细胞因子产生,并强调它们在GVHD中的可能作用。在37°C和5次刺激4个正常足月分娩和10个正常献血者的肝素化HUCB样品4小时。在存在布雷菲丁的情况下,用佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)和碘霉素处理的%CO2。之后,将细胞用不同组合的CD3,CD4或CD8染色。最后,细胞透化后,将细胞用Il-2,Il-4或IFN-γ染色。在FACScan流式细胞仪上进行数据采集,与APB相比,HUCB T淋巴细胞产生的Il-2,Il-4和IFN-γ更少。在HUCB中,II-2,Il-4和IFN-γ主要由CD4 + T细胞产生。在APB中,与CD8 + T淋巴细胞相比,CD4 +细胞也主要产生Il-2和Il-4,但是,CD4 +和CD8 + T细胞都产生IFN-γ。这些结果表明,APB和HUCB中T细胞之间的细胞因子谱存在明显差异。

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