Lee Eunkyung, Min Hae-Ki, Oskeritzian Carole A, Kambe Naotomo, Schwartz Lawrence B, Wook Chang Hyeun
College of Pharmacy, Yeungnam University, Gyongsan 712-749, South Korea.
Cell Immunol. 2003 Nov;226(1):30-6. doi: 10.1016/j.cellimm.2003.11.001.
We previously reported that rhIL-4 induced apoptosis and rhIL-6 mediated protection of human mast cells derived from cord blood mononuclear cells. Based on the result, we attempted to obtain the phenotypes and differentiation of CD3+ cells from cord blood by investigating their cell surface markers in the presence of rhSCF plus rhIL-4. The effect of co-cultured CD3+ cells on fetal liver mast cells (FLMCs) was also determined. Phenotypes from cord blood-derived cells were analyzed by flow cytometry and cell numbers were determined. Fetal liver mast cells were cultured with cord blood-derived cells (mainly CD3+) in the presence of rhSCF and/or rhIL-4 and were analyzed to determine cell number and expression of Kit+ and FcepsilonR1. The percentage of CD3+ cells from cord blood-derived cells on day 0 was about 41 +/- 13.5%, following monocytes and granulocytes. CD3+ cells increased in number (1.5-fold) and purity (90%), whereas other cell types did not survive. More than 60% of CD3+ cells from cord blood at day 0 were CD4(-)CD8-. These double-negative cells dramatically decreased by 1 week of culture, while CD4+CD8+ cells increased in number and purity through 3 weeks of culture, and then decreased as greater numbers of single-positive T cells emerged. We also found that FcepsilonR expression on FLMC increased in the presence of rhIL-4, but was not affected by the T cells that developed from cord blood mononuclear cells. The results indicate that IL-4, a Th2 type cytokine, together with rhSCF, can induce T cell proliferations, differentiation, and maturation from cord blood progenitor cells.
我们之前报道过，重组人白细胞介素-4（rhIL-4）可诱导凋亡，而重组人白细胞介素-6（rhIL-6）可介导对源自脐血单个核细胞的人肥大细胞的保护作用。基于该结果，我们试图通过在重组人干细胞因子（rhSCF）加rhIL-4存在的情况下研究其细胞表面标志物，来获得脐血中CD3⁺细胞的表型和分化情况。还测定了共培养的CD3⁺细胞对胎儿肝脏肥大细胞（FLMCs）的影响。通过流式细胞术分析脐血来源细胞的表型，并确定细胞数量。在rhSCF和/或rhIL-4存在的情况下，将胎儿肝脏肥大细胞与脐血来源细胞（主要是CD3⁺细胞）共培养，并进行分析以确定细胞数量以及Kit⁺和FcepsilonR1的表达。第0天脐血来源细胞中CD3⁺细胞的百分比约为41±13.5%，仅次于单核细胞和粒细胞。CD3⁺细胞数量增加（1.5倍）且纯度提高（90%），而其他细胞类型无法存活。第0天脐血中超过60%的CD3⁺细胞为CD4⁻CD8⁻。这些双阴性细胞在培养1周后显著减少，而CD4⁺CD8⁺细胞数量和纯度在培养3周内增加，随后随着更多单阳性T细胞出现而减少。我们还发现，在rhIL-4存在的情况下，FLMC上的FcepsilonR表达增加，但不受脐血单个核细胞发育而来的T细胞影响。结果表明，Th2型细胞因子白细胞介素-4与rhSCF一起，可诱导脐血祖细胞的T细胞增殖、分化和成熟。
