肿瘤导致的 T 房间的 graft-versus-tumor (GVT ) 效果在 vitro 的搏动抗原的 CD8+ 树枝状的房间(DC ) 在这研究被调查。不成熟的 CD8+ DC 被使用 cytokine 鸡尾酒从 C57BL/6 (H-2b ) 骨头髓房间准备。在文化的第三天,CD8+ DC 被 allogeneic 搏动( Balb/c , H-2d ) EL9611 白血病抗原,或 RM-1 syngeneic 前列腺癌症抗原与 0 的集中系列, 2.5 , 5.0 , 10.0 , 20.0 g/mL ,分别地然后装载抗原的不成熟的CD8+ DC 根据 1:1 , 2:1 和 4:1 的 DC/T 比率与 syngeneic T 房间是co有教养的。T 房间增长被 MTT 试金测量。包括干扰素 gamma (IFN-) 和在 CD8+ DC 和 T 合作文化上层清液的 interleukin-10 (IL-10 ) 的 Cytokines 被使用 ELISA 检测。抗原特定的 T 房间的细胞毒素的效果被 LDH 版本试金测试。使用刺激因素(GM-CSF ) 和 interleukin-4 (IL-4 ) 的 granulocyte 巨噬细胞殖民地从 C57BL/6 (H-2b ) 骨头髓房间导致的常规成熟 DC (mDCs ) 用作控制。结果证明 T 房间的 proliferative 活动由与 allogeneic 装载的 CD8+ DC 刺激了或 syngeneic 肿瘤抗原被扩充, CD8+ DC/T 比率增加了(P < 0.05 ) 。什么时候抗原集中 5 g/mL 和 CD8+ DC/T 比率 2:1, CD8+ 的能力刺激 T 房间增长的 DC 比在 allogeneic 肿瘤的 mDC 控制高搏动抗原的组(P < 0.05 ) ,然而并非在 syngeneic 肿瘤搏动抗原的组(P > 0.05 ) 。在CD8+ DC 和 T 房间合作文化上层清液的 IFN-10 和 IL-10 的水平在 allogeneic 和 syngeneic 被增加搏动抗原的组( P < 0.05 ),并且 cytokine 水平在 allogeneic 是更高的当CD8+ DC/T 是 1:1 或 2:1 时,比在 syngeneic ,抗原组织的搏动抗原的组( P < 0.05 )。在那里存在在 IL-10 和 T 房间增长的水平之间的否定关联。T 房间 cytotoxicity 试金证明当 CD8+ DC 与 allogeneic 肿瘤抗原被搏动时,杀死效率的最大的 T 房间能到达(100
The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α+ dendritic cells (DCs) in vitro was investigated in this study. Immature CD8c(DCs were prepared from C57BL/6 (H-2b) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α+ DCs were pulsed by allogeneic (Balb/c, H-2d) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α+ DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α+ DCs and T co-culture supernatant were detected by using ELI SA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced, from C57BL/6 (H-2b) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α+ DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8& DC/T ratio increased (P〈0.05). When antigen concentration 〈 5 μg/mL and CD8a+ DC/T ratio 〈 2:1, the ability of CD8α+ DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P〈0.05), but not in syngeneic tumor antigen-pulsed groups (P〉0.05). The level of IFN-γ and IL-10 in CD8α+DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P〈0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α+ DC/T was 1:1 or 2:1 (P〈0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxi