树突状细胞激活的肿瘤浸润性淋巴细胞体外特异性抗小鼠肝癌研究

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目的树突状细胞(DC)是目前已知的功能最强的抗原提呈细胞(APC),可以向包括肿瘤浸润性淋巴细胞(TIL)在内的T淋巴细胞提呈抗原,并诱发细胞毒T淋巴细胞(CTL)反应。本文旨在探讨H22细胞和B16细胞全细胞性抗原致敏的树突状细胞激活的肿瘤浸润性淋巴细胞体外抗小鼠肝癌活性。方法从小鼠四肢长骨骨髓中获取DC,应用粒/巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)和肿瘤全细胞性抗原致敏DC,然后用DC激活TIL,观察TIL在体外对H22细胞、Hepal-6细胞和B16细胞的杀伤活性。结果经H22细胞全细胞性抗原致敏的DC激活的TIL具有很高的对H22细胞杀伤活性,杀伤率为(71·31±3·11)%,明显高于其对Hepal-6和B16细胞的杀伤活性[杀伤率分别为(50·11±3·03)%,(30·31±2·89)%];也明显高于未经H22细胞全细胞性抗原致敏的DC激活的TIL、DC激活的脾淋巴细胞和未经DC激活的脾淋巴细胞对H22细胞杀伤活性[杀伤率分别为(49·80±3·21)%,(48·76±3·60)%和(19·23±2·71)%]和对Hepal-6细胞杀伤活性[杀伤率分别为(39·4±3·21)%,(38·62±2·87)%和(18·73±2·40)%]以及对B16细胞杀伤活性[杀伤率分别为(26·38±2·51)%,(25·82±2·70)%和(18·34±3·01)%],同时经B16细胞全细胞性抗原致敏的DC激活的TIL(来源于H22瘤体)也可诱导相对较低的对B16细胞的特异性细胞杀伤活性。结论来源于H22瘤体的TIL经H22细胞全细胞性抗原致敏的DC激活后可产生很强的针对H22细胞的特异性杀伤活性,明显高于其他各组,说明DC能诱导TIL产生高效而特异的体外抗小鼠肝癌免疫。 Objective Dendritic cells (DCs) are currently the most potent antigen-presenting cells (APCs) that can present antigens to T lymphocytes including tumor infiltrating lymphocytes (TILs) and induce cytotoxicity T lymphocyte (CTL) reaction. This article aims to investigate the anti-mouse hepatocarcinoma activity of tumor-infiltrating lymphocytes activated by dendritic cells activated by whole cell antigen of H22 cells and B16 cells. Methods DCs were harvested from the long bone marrow of the extremities of mice and sensitized DCs with granulocyte / macrophage colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor cell-specific antigen The killing activity of TIL on H22 cells, Hepal-6 cells and B16 cells in vitro. Results TIL activated by DCs sensitized by H22 cells had a high cytotoxicity against H22 cells (71 · 31 ± 3.11%), which was significantly higher than that of HIL cells. (50.11 ± 3.03%, (30.31 ± 2.89)%], respectively, which was also significantly higher than that of DCs activated by whole cell-free antigen of H22 cells , The cytotoxic activity of DC-activated splenic lymphocytes and non-DC-activated splenic lymphocytes against H22 cells [(49.80 ± 3.21)%, (48.76 ± 3.60)%, and · 23 ± 2 · 71%] and cytotoxicity against Hepal-6 cells (killing rate were 39.4 ± 3.21%, 38.62 ± 2.87% and 18.73 ± 2 · 40%] and the killing activity of B16 cells (killing rate were (26.38 ± 2.51)%, (25.82 ± 2.70)% and (18.34 ± 3.101)% respectively] DC-activated TIL (derived from H22 tumor) sensitized with B16-cell whole-cell antigen also induced a relatively low specific cytotoxic activity on B16 cells. Conclusions The TILs derived from H22 tumor cells are highly specific for H22 cells when activated by DCs sensitized by whole cell antigen of H22 cells, which is significantly higher than that of other groups, indicating that DC can induce efficient TIL production Specific in vitro anti-mouse liver cancer immunity.
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