Asada-Mikami Rumiko, Heike Yuji, Harada Yukie, Kanai Sachiyo, Ikarashi Yoshinori, Kato Kazunori, Shirakawa Kazuo, Takaue Yoichi, Abe Tatsuo, Wakasugi Hiro
Pharmacology Division, National Cancer Center Research Institute and Hematopoietic Stem Cell Transplant/Immunotherapy Division, National Cancer Center Hospital, Chuo-ku, Tokyo 104-0045, Japan.
Cancer Sci. 2003 Apr;94(4):383-8. doi: 10.1111/j.1349-7006.2003.tb01451.x.
In the present study, unpurified peripheral blood mononuclear cells (PBMCs) from various sources, including steady-state blood (normal donors) and granulocyte colony-stimulating factor (G-CSF)-mobilized blood (cancer patients and normal donors) (G-PBSC), were cultured in RPMI-1640 in the presence of IL-2 and alpha-galactosylceramide (alpha-GalCer) to expand V alpha 24(+) T cells, and their expansion kinetics were compared. G-CSF-mobilized cells showed markedly higher expansion potential (350-fold expansion of V alpha 24(+) T cells, regardless of whether the cells were from cancer patients or normal donors) than steady-state cells (15-fold expansion, compared to the initial inoculums) (n = 5, P < 0.01). We also confirmed that the CD14(-) fraction of G-PBSCs contained a large number of precursors of V alpha 24(+) T cells, compared to PBSCs, as well as a large number of CD14(+) cells, which assist V alpha 24(+) T cell proliferation. Our simple and practical procedure, which eliminates complicated cell manipulation (including cell purification), produces efficient expansion of V alpha 24(+) T cells when G-CSF-mobilized blood cells are cultured with alpha-GalCer.
在本研究中，将来自各种来源的未纯化外周血单个核细胞（PBMC），包括稳态血液（正常供体）和粒细胞集落刺激因子（G-CSF）动员的血液（癌症患者和正常供体）（G-PBSC），在含有IL-2和α-半乳糖神经酰胺（α-GalCer）的RPMI-1640培养基中培养以扩增Vα24(+) T细胞，并比较它们的扩增动力学。与稳态细胞（相对于初始接种物扩增15倍）相比，G-CSF动员的细胞显示出明显更高的扩增潜力（Vα24(+) T细胞扩增350倍，无论细胞来自癌症患者还是正常供体）（n = 5，P < 0.01）。我们还证实，与PBSC相比，G-PBSC的CD14(-)部分含有大量Vα24(+) T细胞前体，以及大量辅助Vα24(+) T细胞增殖的CD14(+)细胞。当用α-GalCer培养G-CSF动员的血细胞时，我们简单实用的方法（消除了复杂的细胞操作，包括细胞纯化）可有效扩增Vα24(+) T细胞。
