Impact of culture medium on the expansion of T cells for immunotherapy
Impact of culture medium on the expansion of T cells for immunotherapy
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DOI：
10.3109/14653240903219114
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发表时间：
2009-01-01
期刊：
影响因子：
4.5
通讯作者：
Kakimi, Kazuhiro
中科院分区：
文献类型：
--
作者：
Sato, Keisuke;Kondo, Makoto;Kakimi, Kazuhiro
癌症免疫治疗的临床益处的令人鼓舞的证据开始在几个临床试验中积累。癌症免疫疗法基于两种主要方法，主动疫苗接种和细胞转移疗法。T细胞的体外扩增需要监测疫苗诱导的抗原特异性T细胞或制备大量的反应性淋巴细胞过继transfer.Methods我们研究了培养基对T细胞生长，细胞毒性和表型的影响后，使用固定化抗CD 3单克隆抗体或唑来膦酸盐刺激激活。结果RPMI、AIM-V、OpTmizer 3种培养液中加入足够的自体血清后，对抗CD 3刺激后的T细胞扩增效果良好。向AIM-V或OpTmizer中添加自体血清可使获得的细胞数量增加至相似程度，但其表型和功能完全不同。与OpTmizer一起培养的活化T细胞比任何其他培养物介导更大的细胞毒性。无论使用哪种培养基，CD 3刺激后扩增的主要群体是CD 3（+）CD 8（+）。在RPMI和AIM-V中诱导更多的CD 3（+）CD 4（+）T细胞，而在OpTmizer中诱导更多的CD 3（-）CD 56（+）细胞和CD 3（+）CD 56（+）T细胞。当细胞刺激唑来膦酸盐14天，约7.2倍和11.5倍以上的γ δ T细胞获得在OptiTmizer比AIM-V或RPMI.Conclusions成功的免疫疗法依赖于选择合适的培养基，以支持所需的T细胞类型的有效扩增。
Background aims Encouraging evidence of clinical benefits from cancer immunotherapy is beginning to accumulate in several clinical trials. Cancer immunotherapy is based on two main methods, active vaccination and cell-transfer therapy. The ex vivo expansion of T cells is required to monitor vaccine-induced antigen-specific T cells or prepare large numbers of reactive lymphocytes for adoptive transfer.Methods We examined the influence of culture medium on T-cell growth, cytotoxicity and phenotype after activation using immobilized anti-CD3 monoclonal antibody or Zoledronate stimulation. Peripheral blood mononuclear cells (PBMC) were cultured in RPMI, AIM-V or OpTmizer with or without autologous serum.Results When supplemented with sufficient serum, RPMI was a good culture medium for T-cell expansion following anti-CD3 stimulation. Addition of autologous serum to AIM-V or OpTmizer increased the numbers of cells obtained to a similar extent, but their phenotype and function were quite different. Activated T cells cultured with OpTmizer mediated greater cytotoxicity than any other culture. Regardless of the media used, the main population expanded after CD3 stimulation was CD3(+) CD8(+). While more CD3(+) CD4(+) T cells were induced in RPMI and AIM-V, more CD3(-) CD56(+) cells and CD3(+) CD56(+) T cells were induced in OpTmizer. When cells were stimulated by Zoledronate for 14 days, approximately 7.2 times and 11.5 times more gamma delta T cells were obtained in OpTmizer than AIM-V or RPMI, respectively.Conclusions Successful immunotherapy depends on the selection of appropriate culture media to support efficient expansion of the type of T cell desired.