树突状细胞对肿瘤浸润淋巴细胞抗小鼠肝癌活性影响的研究  

Study of Anti-mouse Hepatocellular Carcinoma Activities of TIL Affected by DC

作　　者：刘剑勇[1] 张志明[1] 赵荫农[1] 张力图[1] 张春燕[1] 唐凯[1] 吴飞翔[1] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆乳腺外科,530021

出　　处：《实用癌症杂志》2006年第3期232-237,共6页The Practical Journal of Cancer

基　　金：广西科学研究与技术开发计划资助项目(编号:桂科攻0143052)

摘　　要：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-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及抑瘤作用。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、血清TNF活性、抑瘤作用以及瘤体病理改变,并与对照组相比较。结果①H22-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)%,(30.31±2.89)%],也明显高于未经DC激活的TIL、H22-DC-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(39.40±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。②H22-DC-TIL可明显诱导提高荷瘤脾淋巴细胞NK、LAK和CTL活性[活性为(30.43±1.35)%、(31.40±1.80)%、(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/ml],它们均达正常对照组水平,与未经DC激活的TIL组、H22-DC-脾淋巴细胞组、未经DC激活的脾淋巴细胞组、生理盐水组分别对应比较,差异均有显著性(P<0.01)。该组瘤体内淋巴细胞浸润程度也高于对照组,其瘤体生长明显受到抑制。结论①H22-DC-TIL可产生很强的体外针对H22细胞的特异性杀伤活性。②H22-DC-TIL具有很强的特异性抗小鼠肝癌作用。Objective To investigate the killing activity of H22-DC-TIL[tumor infiltrating lymphocytes （TIL） stimulated by dendritic cells （DC） that were sensitized by H22 cell antigen] on carcinoma cells in vitro and to explore the influence of immune and inhibition of the progression of tumor after adoptive immunotherapy with H22-DC-TIL on mice bearing tumor. Methods DC were isolated from mouse bone marrow and sensitized by granulocyte/macrophage colony stimulating factor （GM-CSF）, interleukin-4 （IL-4） and tumor antigen. Then TIL were stimulated by DC and their killing activity on H22, Hepal-6 and B16 cells were observed in vitro. The activities of splenocyte NK, LAK, CTL, serum TNF, inhibition of the progression of tumor and pathologic change of tumor of mice bearing tumor were assayed respectively after treated with H22-DC-TIL,and compared with control groups. Results ① H22-DC-TIL had very high killing activity on H22 cells [killing rate： （71.31 ± 3.11） % ], it was higher killing activity than those on Hepal-6 and B16 cells [killing rates： （50.11 ± 3.03） %, （30.31 ± 2.89） % ,respectively] ,and it also was much higher killing activity than those of TIL not stimulated by DC and splenocytes stimulated by DC as well as splenocytes not stimulated by DC on H22 cells [killing rates： （49.80 ± 3.21）%, （48.76 ± 3.60）%, （19.23 ± 2.71）% ,respectively] ,on Hepal-6 cells [killing rates： （39.40 ± 3.21 ） %, （38.62 ± 2.87） %, （ 18.73 ± 2.40） % , respectively ] and on B 16 cells [ killing rates： （26.38 ± 2.51 ） %, （25.82 ± 2.70） %, （18.34 ± 3.01）% ,respectively] ,B16-DC-TIL [TIL stimulated by DC that were sensitized by B16 cell antigen, and the TIL came from H22 tumor] had relative lower killing activity on B16 cells. ② The activities of splenocyte NK,LAK ,CTL and serum TNF of mice bearing tumor can be induced to enhance＇obviously by H22-DC-TIL ,they reached the level of normal group,and had significant difference comparing with TIL not stimulated

关 键 词：树突状细胞 肿瘤浸润淋巴细胞 H22细胞 杀伤活性 小鼠肝癌 

分 类 号：R73-36[医药卫生—肿瘤]