[Effects of Sema3A derived from tumor cells on functions of dendritic cells].
[Effects of Sema3A derived from tumor cells on functions of dendritic cells].
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发表时间：
2010-07
影响因子：
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通讯作者：
Xie-lai Zhou;Yin Huang;Fang Wang;L. Cai;Li-huang Zhang;Li-yun Shi
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作者：
Xie-lai Zhou;Yin Huang;Fang Wang;L. Cai;Li-huang Zhang;Li-yun Shi
目的探讨肿瘤细胞源性Sema3A对小鼠树突状细胞（dc）免疫功能的影响。方法用小干扰RNA、Si-Sema和Si-mut转染肺腺癌A549细胞，采用实时荧光定量PCR和Western-blot检测干扰效率。将培养的肿瘤细胞、Si-Sema和si - muta感染的肿瘤细胞的浓缩上清液分别进行DCs处理。通过流式细胞术分析DCs的免疫表型、IL-12P70的产生和DCs刺激DO11的能力。采用酶联免疫吸附法（ELISA）检测10个T细胞分泌ifn - γ和IL-2的情况。结果与Si-mut细胞相比，敲低Si-Sema3A可显著降低A549细胞的Sema3A分泌。暴露于Si-Sema细胞上清液的DCs显示MHC、CD40和CD80水平升高，IL-12P70的产生增加，激活抗原特异性T细胞的能力增强，ifn - γ和IL-2水平显著升高。结论A549细胞分泌Sema3A抑制dc的成熟和功能，这可能与肿瘤细胞逃避免疫的机制有关。
OBJECTIVE To investigate the effects of tumor cell-derived Sema3A on the immunological functions of murine dendritic cells (DCs). METHODS Lung adenocarcinoma A549 cells were transfected with small interference RNA, Si-Sema and Si-mut, and the interference efficiency was determined by real-time PCR and Western-blot. The concentrated supernatants from cultured tumor cells, Si-Sema and Si-mut-infected tumor cells were subjected to DCs respectively. The immunophenotypes of DCs were analyzed by flow cytometry, the production of IL-12P70 and the ability of DCs to stimulate DO11. 10 T cells secreting IFN-gamma and IL-2 were detected by enzyme linked immunosorbent assay (ELISA). RESULTS Knockdown with Si-Sema3A significantly decreased the secretion of Sema3A by A549 cells in comparison with the Si-mut cells. DCs exposed to supernatants from Si-Sema cells showed elevated levels of MHC, CD40 and CD80, more production of IL-12P70, and enhanced capability of activating antigen-specific T cells, as evidenced by the remarkably increased levels of IFN-gamma and IL-2. CONCLUSION A549 cells secrete Sema3A to inhibit the maturation and functions of DCs, which might be associated with the unidentified mechanism of immune evasion by tumor cells.