论文部分内容阅读
为探讨凋亡Daudi细胞负载的树突状细胞 (DC )激发诱导肿瘤抗原特异性CTL及其生物学特性 ,采用常规方法从人外周血单个核细胞 (PBMC )诱导DC ,激发型CD40mAb联合 50Gyγ射线辐照诱导Daudi凋亡后负载DC ,然后与自体T细胞共育 ,并联合IL 2激发诱导肿瘤特异性CTL ;采用免疫荧光标记和流式细胞仪测定分析膜分子的表达 ;ELISA检测细胞因子的产生 ;Dextran FITC内吞实验分析DC的抗原摄取能力 ;3H掺入试验和51 Cr释放试验分别测定CTL的增殖和细胞毒效应。结果 :(1 )细胞因子序贯体外诱导 7d的DC ,对Dextran吞噬能力最强 ;(2 )CD40mAb诱导联合γ射线辐照 ,能有效介导Daudi细胞的凋亡 ;(3 )抗原负载联合CD40mAb激发可使DC上调表达CD1a、CD80、CD86和HLA DR ,并能促进IL 1 2的分泌 ;(4)凋亡Daudi负载后的DC在激发型CD40mAb作用下 ,能激发和扩增对Daudi细胞具有高效和特异杀伤作用的细胞毒性T细胞。因而认为凋亡肿瘤细胞负载联合CD40mAb刺激的DC可有效激活和扩增肿瘤特异性CTL。
To investigate the tumor antigen-specific CTL induced by dendritic cells (DCs) loaded with apoptotic Daudi cells and its biological characteristics, DCs were induced from human peripheral blood mononuclear cells (PBMCs) by conventional methods. The stimulated CD40 mAbs in combination with 50 Gy γ-rays Daudi cells were loaded with Daudi after irradiation, co-cultured with autologous T cells and IL-2-stimulated tumor-specific CTLs. The expression of membrane molecules was analyzed by immunofluorescence and flow cytometry. The expressions of cytokines The Dextran FITC endocytosis assay was used to analyze the antigen uptake capacity of DCs. 3H-incorporation assay and 51 Cr release assay were used to determine the proliferation and cytotoxic effects of CTL. (2) CD40mAb induced γ-ray irradiation can effectively induce apoptosis of Daudi cells; (3) Antigen load combined with CD40 mAb Activation can up-regulate the expression of CD1a, CD80, CD86 and HLA DR, and promote the secretion of IL-12. (4) The apoptotic Daudi-loaded DCs can stimulate and amplify Daudi cells under the action of CD40mAb Efficient and specific killing of cytotoxic T cells. Therefore, it is considered that apoptotic tumor cells loaded with CD40 mAb-stimulated DCs can effectively activate and amplify tumor-specific CTLs.