中国肿瘤生物治疗杂志2012，Vol.19Issue(1)：22-28,7.DOI:10.3872/j.issn.1007-385X.2012.01.004
树突状细胞感染PSMA/4-1BBL基因重组腺病毒后的免疫功能变化
Changes in immune function of dendritic cells infected by recombinant adenovirus containing PSMA and 4-1BBL genes
摘要
Abstract
Objective: To observe changes in immune function of dendritic cells (DCs) infected by recombinant adeno-virus containing human prostate specific membrane antigen (PSMA) and 4-1BB ligand (4-1BBL) genes. Methods: The empty Ad-GFP infected immature DCs at different multiplicities of infection (MOI) , and the optimal MOI was determined. DCs from peripheral blood of healthy volunteers were infected with recombinant adenoviruses at the optimal MOI. The DCs were allocated into five groups; co-infected group (Ad-PSMA/4-1 BBL-DC) , PSMA-infected group (Ad-PSMA-DC), 4-1 BBL-infected group (Ad-4-1 BBL-DC) , negative control group (Ad-GFP-DC) and non-infected group. Morphological changes of DCs were observed under a fluorescence microscope; and the phenotype changes of CD80, CD83, CD86 of tranfected DCs were detected by FACS, and the expressions of PSMA and 4-1BBL proteins was detected by Western blotting. IL-12 concentrations of DC supernatant in different groups were measured by ELJSA. Allogenic T cell proliferations stimulated with DCs in different groups and the cytotoxic effect on prostate cancer cell lines LNCap, Dul45 and 22RV were analyzed by CCK-8 assay. Results: The optimal MOI was 200. The immunophenotype expressed in the co-infected DC group, such as CD80, CD83, CD86, HLA-DR, was significantly higher than that expressed in the other DCgroups ([38.72 ±0. 99]%, [44.65 ± 0. 77]% , [63. 60 ± 0. 75 ]% , [62. 25 ±0.58]% vs [28.27 ±1.04]%, [28. 08 ±1. 16% ] ,[41.05 ±1.33]% ,[46.87 ±1.12]% ; P<0.05). The secretion of IL-12 in supernatant was greatly higher in the co-infected group than that in single-infected group or non-infected DC group ( [ 134. 29 ± 2. 22 ] ts [79.51 ±1.60], [70.33 ±1.13], [69.67 ±1.43] , [28.88 ±2.97] pg;P<0.05). At the same rate of DCs to T, the co-infected group had a more significant increase in allogenic T cell proliferation than other single-infected group or non-infected group (P<0.05). The cytotoxic rate of PSMA/4-1BBL-DC-CTL against PSMA positive target LNCap cells was significantly stronger than that against PSMA negative target cells such as DU145 and 22RV cells (P<0.05) , and also higher than that of PSMA-DC-CTL and 4-1 BBL-DC-CTL (P < 0.05). Conclusion: Ad-PSMA/4-1 BBL-infected DCs not only own high secretion of IL-12, but also stimulate and enhance the cytotoxic effect of tumor specific CTLs against PSMA positive prostate cancer cells which highly express PSMA protein. In addition, the DCs infected with Ad-PSMA and Ad-4-1 BBL induce more effective tumor specific CTLs than single-infected DCs.关键词
树突状细胞/前列腺特异性膜抗原基因/4-1BBL基因/细胞毒性T淋巴细胞/前列腺癌Key words
dendritic cell/ prostate specific membrane antigenen gene (PSMA)/ 4-1 BBL gene/ cytotoxic T lymphocyte (CTL) / prostate cancer分类
医药卫生引用本文复制引用
杨明花,王帅,孙海燕,刘晓辉,马楠,隋承光..树突状细胞感染PSMA/4-1BBL基因重组腺病毒后的免疫功能变化[J].中国肿瘤生物治疗杂志,2012,19(1):22-28,7.基金项目
辽宁省科技厅科学技术计划项目资助(No.2010225034). （No.2010225034）
