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目的:通过分析小鼠肝癌H22细胞全细胞性抗原致敏小鼠骨髓树突状细胞(DC)前后DC表型变化及其分泌细胞因子的变化,探讨肝癌细胞全细胞性抗原致敏DC激活肿瘤浸润性淋巴细胞(TIL)的抗癌机制。方法:取得小鼠骨髓细胞并诱导生成DC,用冻融法制备的小鼠肝癌H22细胞全细胞抗原致敏,然后用已致敏的DC激活TILs,测定DC致敏前后DC表面抗原CD80、CD86、CD40和MHCⅡ表达变化及DC分泌细胞因子IL-12、IL-2、IFN-γ和TNF-α的水平,评估激活前后TIL对H22细胞的杀伤活性,同时以小鼠脾淋巴细胞作为杀伤对照。结果:致敏后小鼠骨髓DC表面抗原CD80、CD86、CD40和MHCⅡ表达率分别为(57.55±7.32)%、(54.49±14.20)%、(46.79±8.25)%和(53.94±13.94)%,明显高于未致敏DC的(20.01±5.22)%、(24.56±9.08)%、(18.06±5.13)%和(30.24±8.39)%,P值均<0.05;DC上清液中IL-12、IL-2、IFN-γ和TNF-α浓度分别为(80.40±1.33)、(94.67±3.36)、(29.83±1.20)和(75.01±4.10)ρg/mL,明显高于未致敏DC的(19.35±0.99)、(11.25±0.50)、(1.05±0.09)和(2.02±0.27)ρg/mL,P值均为0.000。经致敏后成熟DC激活的TIL对H22细胞杀伤率为(81.80±2.90)%,明显高于未激活TIL、激活或未激活小鼠脾淋巴细胞的(62.64±3.94)%、(47.35±3.40)%和(28.45±2.56)%。结论:H22细胞全细胞抗原致敏DC后,其表型变化及细胞因子分泌增多是其诱导活化TIL抗癌活性的可能机制。
OBJECTIVE: To investigate the changes of DC phenotypes and cytokines secreted by mouse bone marrow dendritic cells (DCs) sensitized by whole cell antigen of mouse hepatoma H22 cells, and to investigate the effect of DCs activated by whole cell antigen of liver cancer cells Anti-cancer mechanism of infiltrating lymphocytes (TILs). Methods: The mouse bone marrow cells were obtained and induced to generate DC. The whole cell antigens of mouse hepatoma H22 cells were prepared by freeze-thaw method. TILs were activated by DCs sensitized. The expression of DC surface antigen CD80, CD86 , CD40 and MHC Ⅱ expression and the levels of IL-12, IL-2, IFN-γ and TNF-α secreting cytokines in DCs were measured before and after activation. The killing activity of TIL on H22 cells was evaluated before and after activation. Meanwhile, splenic lymphocytes were used as control . Results: After sensitized, the expression rates of CD80, CD86, CD40 and MHCⅡ in bone marrow of the mice were (57.55 ± 7.32)%, (54.49 ± 14.20)%, (46.79 ± 8.25)% and (53.94 ± 13.94)%, (20.01 ± 5.22)%, (24.56 ± 9.08)%, (18.06 ± 5.13)% and (30.24 ± 8.39)%, respectively, with a P value of <0.05. The levels of IL-12 in DC supernatant , The concentrations of IL-2, IFN-γ and TNF-α were (80.40 ± 1.33), (94.67 ± 3.36), (29.83 ± 1.20) and (75.01 ± 4.10) ρg / (19.35 ± 0.99), (11.25 ± 0.50), (1.05 ± 0.09) and (2.02 ± 0.27) ρg / mL respectively. The P values were all 0.000. The killing rate of mature TIL-activated TIL to H22 cells after sensitization was (81.80 ± 2.90)%, which was significantly higher than that of inactive TIL (62.64 ± 3.94)%, (47.35 ± 3.40) )% And (28.45 ± 2.56)%. CONCLUSION: The phenotypic changes and the increase of cytokine secretion induced by H22 cell whole-cell antigen are the possible mechanism of its induction of anti-tumor activity of TIL.