SOCS1 受抑的新型肝癌树突状细胞疫苗制备及功效的初步研究

目的探讨抑制SOCS1表达对LIGHT激活的肝癌树突状细胞疫苗效应的作用。方法用重组小鼠粒-巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白细胞介素4(rmIL-4)体外诱导培养小鼠骨髓单核细胞产生骨髓来源树突状细胞(BMDC),体外负载肝癌细胞HepA可溶性抗原,并用LIGHT和SOCS1反义寡核苷酸(AS1)处理DC,制备负载肝癌抗原DC疫苗;流式细胞仪检测疫苗细胞CD40及CD86表达和ELISA测定其IL-12、IL-1分泌水平以确定DC疫苗细胞的成熟度;通过体外细胞毒T淋巴细胞(CTL)活性、淋巴细胞增殖反应和IL-6、TNF-β分泌水平测定分析疫苗细胞的抗肿瘤免疫应答。结果负载肝癌抗原DC疫苗在LIGHT和AS1的双重作用下,疫苗细胞的成熟标志CD40、CD86、IL-1和IL-12增高(P<0.01),能增强诱导CTL活性、刺激淋巴细胞增殖和分泌IL-6、TNF-β能力(P<0.01)。结论抑制SOCS1能提高肝癌DC疫苗的成熟度,并增强疫苗细胞诱导抗肿瘤免疫应答。

Primary Study on Preparation and Efficiency of Neotype Hepatocellular Carcinoma-dendritic Cell Vaccine with SOCS1 Expression Inhibited

OBJECTIVE: To investigate the effect of SOCS1 expression inhibition on hepatocellular carcinoma-dendritic cell vaccine stimulated by LIGHT. METHODS: The dendritic cells (DC) were generated from mouse bone marrow (BMDC) by cultured in medium containing rmGM-CSF and rmIL-4. The vaccine cells were prepared by loaded with hepatocellular carcinoma HepA antigen and further treated with or without LIGHT and SOCS1 antisense oligonucleotide (AS1). For detecting the maturity of the vaccine cells, the expression of the cell's surface molecules CD40 and CD86 were measured by FACS, and IL-12 and IL-1 secretion from the cells were determined by ELISA. And the CTL activity, cellular proliferation, IL-6 and TNF-beta secretion levels of the vaccine-stimulated lymphocytes were also assessed for analyzing the immune response of lymphocyte. RESULTS: CD40 and CD86 expression of the prepared vaccine cells were obviously enhanced by treatment of LIGHT and AS1, and so were IL-12 and IL-1 (P<0.01). It was also observed that CTL activity, cellular proliferation and IL-6 and TNF-beta secreting levels of lymphocytes that were stimulated by the vaccine cells treated with AS1 were notably enhanced. CONCLUSION: Inhibiting SOCS1 can improve the maturation of hepatocellular carcinoma-DC vaccine cell and can increase its inducing ability of anti-cancer immune response.