Saito Anri, Narita Miwako, Yokoyama Ayumi, Watanabe Norihiro, Tochiki Nozomi, Satoh Noriyuki, Takizawa Jun, Furukawa Tatsuo, Toba Ken, Fuse Ichiro, Aizawa Yoshifusa, Shinada Shohji, Takahashi Masuhiro
Laboratory of Hematology and Oncology, Graduate School of Health Sciences, Niigata University, Japan.
J Clin Exp Hematop. 2007 Nov;47(2):61-72. doi: 10.3960/jslrt.47.61.
In order to establish the method of generating powerful gammadelta T cells for anti-tumor immunotherapy, we investigated the effects of monocyte-derived dendritic cells (mo-DCs) on anti-tumor cytotoxicity of expanded gammadelta T cells. Activation of gammadelta T cells co-cultured for 2-3 days with immature or mature mo-DCs was evaluated by CD69 expression and anti-tumor cytotoxicity using two assays : the 5- (and 6-) carboxyfluorescein diacetate, succinimidyl ester-based cytotoxicity assay and the calcein-AM-based Terascan assay. gammadelta T cells were used as effector cells and myeloma cell line (RPMI8226) or chronic myelogenous leukemia blastic crisis cell line (C2F8) were used as target cells. CD69 expression on gammadelta T cells was enhanced by co-culture with both immature and mature mo-DCs in a cell-number-dependent fashion. CD69 expression was enhanced after addition of mo-DCs of either autologous or allogeneic origin. Activation of gammadelta T cells with mo-DCs enhanced anti-tumor cytotoxicity of gammadelta T cells against RPMI8226 and C2F8 in an effector-to-target ratio-dependent manner. Activation of gammadelta T cells by mo-DCs was associated with the enhancement of anti-tumor cytotoxicity of gammadelta T cells. Potent gammadelta T cells activated by mo-DCs were considered to be applicable to an efficient gammadelta T cell-mediated immunotherapy for tumors.
为了建立用于抗肿瘤免疫治疗的高效γδT细胞生成方法，我们研究了单核细胞衍生的树突状细胞（mo-DC）对扩增的γδT细胞抗肿瘤细胞毒性的影响。通过CD69表达以及使用两种检测方法评估γδT细胞与未成熟或成熟mo-DC共培养2-3天的激活情况：基于5-（和6-）羧基荧光素二乙酸琥珀酰亚胺酯的细胞毒性检测和基于钙黄绿素-AM的Terascan检测。γδT细胞用作效应细胞，骨髓瘤细胞系（RPMI8226）或慢性粒细胞白血病急变期细胞系（C2F8）用作靶细胞。γδT细胞与未成熟和成熟mo-DC共培养均以细胞数量依赖的方式增强了CD69的表达。添加自体或同种异体来源的mo-DC后，CD69表达增强。mo-DC激活γδT细胞以效应细胞与靶细胞比例依赖的方式增强了γδT细胞对RPMI8226和C2F8的抗肿瘤细胞毒性。mo-DC激活γδT细胞与γδT细胞抗肿瘤细胞毒性的增强相关。由mo-DC激活的高效γδT细胞被认为适用于高效的γδT细胞介导的肿瘤免疫治疗。
