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目的:探讨H22细胞全细胞性抗原致敏的DC激活的TIL体外抗小鼠肝癌活性;并将H22细胞全细胞性抗原致敏的DC激活的TIL(H22-DC-TIL)过继免疫荷瘤小鼠,研究其对荷瘤小鼠免疫功能的影响。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;检测应用H22-DC-TIL后荷瘤小鼠的脾淋巴细胞的NK、LAK、CTL活性及血清TNF活性,并与对照组相比较。结果:(1)H22-DC-TIL具有很强的对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%和30.31%±2.89%);也明显高于未经DC激活的TIL、H22-DC-小鼠脾淋巴细胞和未经DC激活的小鼠脾淋巴细胞对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39.40%±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B16-DC-TIL(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。(2)H22-DC-TIL可明显诱导提高荷瘤小鼠脾淋巴细胞NK、LAK和CTL活性(活性分别为30.43%±1.35%、31.40%±1.80%和35.30%±1.20%),并可检测到血清TNF水平明显上升[血清TNF为(40.41±1.85)U/m l],它们均达正常对照组水平,与未经DC激活的TIL组、H22-DC-小鼠脾淋巴细胞组、未经DC激活的小鼠脾淋巴细胞组、生理盐水组分别对应比较,差异均有显著性(P<0.01)。结论:(1)H22-DC-TIL可产生很强的体外针对H22细胞的特异性杀伤活性。(2)H22-DC-TIL可明显诱导提高荷瘤小鼠特异性抗肿瘤免疫反应。
OBJECTIVE: To investigate the anti-mouse hepatoma activity induced by DCs activated by whole cell antigen of H22 cells in vitro and to adoptively immunize T22 cells (DCs) activated by H22 cell-full antigen (DCs) Rats, to study its immune function in tumor-bearing mice. 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 whole cell antigen, The killing activity of TIL on H22 cells, Hepal-6 cells and B16 cells was observed in vitro. The NK, LAK and CTL activities and serum TNF activities of splenic lymphocytes of H22-DC-TIL mice were detected and compared with the control Group comparison. Results: (1) H22-DC-TIL had a strong cytotoxic activity against H22 cells (killing rate was 71.31% ± 3.11%), which was significantly higher than that of Hepal-6 and B16 cells (lethal rates were 50.11 % ± 3.03% and 30.31% ± 2.89%, respectively); also significantly higher than that of TIL, H22-DC-mouse splenic lymphocytes without DC activation and splenic lymphocytes without DC activation (49.80% ± 3.21%, 48.76% ± 3.60% and 19.23% ± 2.71%, respectively) and Hepa-6 cell killing activity (killing rates were 39.40% ± 3.21%, 38.62% ± 2.87% and 18.73% ± 2.40 %) And B16 cell killing activity (killing rate were 26.38% ± 2.51%, 25.82% ± 2.70% and 18.34% ± 3.01% respectively), while the B16-DC-TIL Lower specific cytotoxic activity to B16 cells. (2) The activities of NK, LAK and CTL of splenic lymphocytes were significantly increased in H22-DC-TIL mice (activity of 30.43% ± 1.35%, 31.40% ± 1.80% and 35.30% ± 1.20%, respectively) The level of serum TNF was significantly increased (serum TNF was (40.41 ± 1.85) U / ml], which reached the level of normal control group. Compared with TIL group without DC activation, H22-DC-mouse spleen lymphocyte group There was significant difference (P <0.01) between DC-activated splenic lymphocyte group and normal saline group. Conclusion: (1) H22-DC-TIL can produce a strong specific in vitro activity against H22 cells. (2) H22-DC-TIL can obviously enhance the specific anti-tumor immune response in tumor-bearing mice.