体外刺激人脐带血(HUCB)和成人外周血(APB) T淋巴细胞胞内细胞因子生成的比较

J. G. García Vela, I. Delgado, R. Bornstein, B. Alvarez, M. C. Auray, I. Martín, F. Oña, F. Gilsanz, J. Antônio, García Vela
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引用次数: 12

摘要

迄今为止,不同中心报道了400多例hub移植。有研究表明,与骨髓移植相比,hub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一个指标,因此确定APB和hub在产生这些可溶性因子方面的差异是很重要的。我们的目的是分析hub和APB T淋巴细胞细胞内细胞因子的产生,并强调它们在GVHD中的可能作用。在brefeldine存在的情况下,用phorbol 12‐肉豆酸酯13‐乙酸酯(PMA)和Ionomycin在37°C和5% CO2下刺激8例正常足月分娩和10例正常献血者的肝素化HUCB样本4小时。然后用不同组合的CD3、CD4或CD8染色细胞。最后,在细胞渗透后,用Il - 2、Il - 4或IFN - γ对细胞进行染色。数据采集在FACScan流式细胞仪上进行。与APB相比,HUCB T淋巴细胞产生较少的Il - 2、Il - 4和IFN - γ。在hub中,Il - 2、Il - 4和IFN - γ主要由CD4+ T细胞产生。在APB中,与CD8+ T细胞相比,Il‐2和Il‐4也主要由CD4+细胞产生,然而,IFN‐γ由CD4+和CD8+ T细胞产生。这些结果表明,APB和HUCB中T细胞的细胞因子谱存在明显差异。
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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)
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.
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