Harada Hideki, Saijo Kaoru, Watanabe Satoru, Tsuboi Koji, Nose Tadao, Ishiwata Isamu, Ohno Tadao
RIKEN Cell Bank, RIKEN (The Institute of Physical and Chemical Research), Tsukuba Science City 305-0074, Japan.
Jpn J Cancer Res. 2002 Mar;93(3):313-9. doi: 10.1111/j.1349-7006.2002.tb02174.x.
An anchorage-dependent Wilms tumor cell line HFWT was found to stimulate selective and remarkable expansion of human natural killer (NK) cells from human peripheral blood mononuclear cells (PBMC). After PBMC of healthy donors were cultured on irradiated HFWT cells for 10 - 21 days, the lymphocytes expanded 58- to 401-fold. This NK cell expansion required direct contact of PBMC with live, but not fixed, HFWT cells. The PBMC from an end-stage brain tumor patient also expanded 156-fold, whereas those cultured with irradiated NK-sensitive K562 grew only 30.5-fold. CD16+ CD56+ NK cells accounted for more than 70% of the population expanded on HFWT cells. No essential difference in expression of NK receptors was observed in the expanded NK cells on HFWT and K562 and without feeder cells. The expanded NK cells killed not only fresh HFWT cells but, unexpectedly, also MHC class I-expressing autologous brain tumor cells at an effector/target ratio of 4 for 24 h. These results will contribute to the development of a large-scale preparation method for human NK cells, which will aid studies of NK cell biology and possible treatment of brain tumors.
研究发现,一种锚定依赖型肾母细胞瘤细胞系HFWT可刺激人外周血单个核细胞(PBMC)中的人自然杀伤(NK)细胞选择性且显著地扩增。健康供体的PBMC在经辐照的HFWT细胞上培养10至21天后,淋巴细胞扩增了58至401倍。这种NK细胞扩增需要PBMC与活的而非固定的HFWT细胞直接接触。一名晚期脑肿瘤患者的PBMC也扩增了156倍,而与经辐照的NK敏感型K562细胞共同培养的PBMC仅扩增了30.5倍。在HFWT细胞上扩增的细胞群体中,CD16 + CD56 + NK细胞占比超过70%。在HFWT和K562细胞上以及无饲养细胞情况下扩增的NK细胞中,未观察到NK受体表达存在本质差异。扩增后的NK细胞不仅能杀伤新鲜的HFWT细胞,而且出乎意料的是,在效应细胞与靶细胞比例为4的情况下,24小时内还能杀伤表达MHC I类分子的自体脑肿瘤细胞。这些结果将有助于开发一种大规模制备人NK细胞的方法,这将有助于NK细胞生物学研究以及脑肿瘤的可能治疗。