García Vela J A, Delgado I, Bornstein R, Alvarez B, Auray M C, Martin I, Oña F, Gilsanz F
Department of Hematology, Hospital Universitario de Getafe, Madrid, Spain.
Anal Cell Pathol. 2000;20(2-3):93-8. doi: 10.1155/2000/572952.
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 degrees C and 5% CO2 with phorbol 12-myristate 13-acetate (PMA) and lonomycin 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-gamma. Data acquisition was performed on a FACScan flow cytometer. Compared to APB, HUCB T lymphocytes produced less Il-2, Il-4 and IFN-gamma. In HUCB, Il-2, Il-4 and IFN-gamma 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-gamma 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.
迄今为止,不同中心已报告了400多例脐血移植。有人提出,与骨髓移植相比,脐血移植的移植物抗宿主病(GVHD)有所减少。由于细胞产生细胞因子是细胞功能的一种指标,因此确定成人外周血(APB)和脐血在这些可溶性因子产生方面的差异很重要。我们的目的是分析脐血和成人外周血T淋巴细胞的细胞内细胞因子产生情况,并强调它们在移植物抗宿主病中的可能作用。来自8例正常足月分娩和10例正常献血者的肝素化脐血样本,在37℃、5%二氧化碳条件下,用佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)和离子霉素在布雷菲德菌素存在的情况下刺激4小时。之后,细胞用不同组合的CD3、CD4或CD8进行染色。最后,在细胞通透后用白细胞介素-2(Il-2)、白细胞介素-4(Il-4)或γ干扰素(IFN-γ)进行染色。在FACScan流式细胞仪上进行数据采集。与成人外周血相比,脐血T淋巴细胞产生的Il-2、Il-4和IFN-γ较少。在脐血中,Il-2、Il-4和IFN-γ主要由CD4+T细胞产生。在成人外周血中,与CD8+T淋巴细胞相比,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,成人外周血和脐血中的T细胞在细胞因子谱方面存在明显差异。