联合过继免疫细胞输注小鼠后DC、NK分布的研究

The Localization of Natural Killer Cells and Dendritic Cells in Murine Transplanted Tumor after Joint Adoptive-Immunity Cell Infusion

目的:观察自然杀伤(NK)细胞/树突状细胞(DC)联合过继免疫治疗小鼠皮下黑色素瘤时效应细胞在荷瘤小鼠体内的分布。方法:建立C57BL/6小鼠皮下黑色素瘤模型,体外培养增殖小鼠NK细胞、DC并标记,以不同比例混合两种细胞,分别以静脉注射和瘤内注射的方式回输荷瘤小鼠体内,于注射后不同时间点取肿瘤、肝、脾、肺组织进行切片,观察各组织中效应细胞的数量及分布。结果:静脉注射组NK细胞、DC主要分布在肿瘤微循环及实质组织中,各时间点NK细胞、DC的浸润数量有显著差异(P<0.01),注射后4h浸润的效应细胞最多,12h最少,在脾脏中见少量NK细胞的分布,在肝脏及肺组织中偶见。瘤内注射组NK细胞、DC主要分布在肿瘤实质组织中,各观察时间点NK细胞、DC的浸润数量有显著差异(P<0.01),注射后1h浸润的效应细胞最多,12h最少,在肝、脾、肺组织中未见NK细胞、DC分布。两种注射方式下按不同比例混合进行NK细胞、DC的过继治疗,各组间差异具有显著性(P<0.01),混合注射组效应细胞在肿瘤组织内的浸润多于单独注射组。结论:NK细胞、DC免疫过继后能够“靶向性”聚集在肿瘤周围及肿瘤内部,对肿瘤具有治疗作用,NK细胞、DC联合过继免疫治疗效果优于单独注射,提示NK细胞、DC联合输注可能成为肿瘤生物治疗的新模式。

Objective： To study the localization of effector adoptively transferred natural killer （NK） cells and dendritic cells （DCs） in murine subcutaneously tumor. Methods： B16-F10 melanoma cell line was implanted subcutaneously into C57BL/6 mice. NK cells and DCs were prepared and labeled with Brdu and DAPI. The mice received labeled NK cells and DCs via the vein or located injection. Tumor and other organs were removed at different time points after the adoptive transfer and processed. The histopathological change of tumors were observed by HE staining. The labeled NK cells and DCs were obsevered and numbered by use of fluorescense microscope. Results： NK cells and DCs adopted by. the vein injection accumulated mostly in tumors and microcirculation of tumor, and the number of effector cells was significantly different （P〈0.01）. By four hours after injection, significantly more cells were seen in the tumor compared with other time points, and by 12 hours after injection, that was the least. As in the cases of adoptived by localized injection, infiltrative NK cells and DCs localized in the tumors. And the number of the effective cells in different time points was significantly different （P〈0.01）. It was the highest in one hour and the lowest in twelve hours after injection. There was a significantly difference （P〈0.01） as adopting different cells, and the number of received NK cells combined with DCs was higher than injection of single kind of cells. Conclusions： NK cells and DCs are accumulated in tumor and those lead to the necrosis or apoptosis of the tumors.NK cells combined with DC are effective immunotherapy in tumor of mouse.