Li Ming, Wang Bin, Wu Zhonghua, Zhang Jiadong, Shi Xiwen, Cheng Wenlan, Han Shuangyin
Department of Neurosurgery, Zhengzhou University People's Hospital, 7 Weiwu Road, Zhengzhou City, Henan Province, 450003, People's Republic of China.
Tumour Biol. 2015 Jul;36(7):5497-503. doi: 10.1007/s13277-015-3217-5. Epub 2015 Feb 13.
Dendritic cells loaded with tumor-associated antigens can effectively stimulate the antitumor immune response of cytotoxic T lymphocytes in the body, which facilitates the development of novel and effective treatments for cancer. In this study, the adenovirus-mediated ephrinA1-PE38/GM-CSF was successfully constructed using the overlap extension method, and verified with sequencing analysis. HEK293 cells were infected with the adenovirus and the cellular expression of ephrinA1-PE38/GM-CSF was measured with an enzyme-linked immunosorbent assay. The recombinant adenovirus was then delivered into the tumor-bearing rats and the results showed that such treatment significantly reduced the volumes of gliomas and improved the survival of the transplanted rats. The results from immunohistochemistry and flow cytometry suggested that this immunomodulatory agent cause activation of dendritic cells. The findings that ephrinA1-PE38/GM-CSF had a high efficacy in the activation of the dendritic cells would facilitate the development of in vivo dendritic-cell vaccines for the treatment of gliomas in rats. Our new method of DC vaccine production induces not only a specific local antitumor immune response but also a systemic immunotherapeutic effect. In addition, this method completely circumvents the risk of contamination related to the in vitro culture of DCs, thus greatly improving the safety and feasibility of clinical application of the DC vaccines in glioma.
负载肿瘤相关抗原的树突状细胞可有效刺激体内细胞毒性T淋巴细胞的抗肿瘤免疫反应，这有助于开发新型有效的癌症治疗方法。在本研究中，采用重叠延伸法成功构建了腺病毒介导的ephrinA1-PE38/GM-CSF，并通过测序分析进行了验证。用该腺病毒感染HEK293细胞，并用酶联免疫吸附测定法检测ephrinA1-PE38/GM-CSF的细胞表达。然后将重组腺病毒导入荷瘤大鼠体内，结果表明这种治疗显著减小了胶质瘤的体积并提高了移植大鼠的存活率。免疫组织化学和流式细胞术结果表明，这种免疫调节剂可激活树突状细胞。ephrinA1-PE38/GM-CSF在激活树突状细胞方面具有高效性这一发现将有助于开发用于治疗大鼠胶质瘤的体内树突状细胞疫苗。我们生产DC疫苗的新方法不仅能诱导特异性的局部抗肿瘤免疫反应，还能产生全身免疫治疗效果。此外，该方法完全规避了与DCs体外培养相关的污染风险，从而大大提高了DC疫苗在胶质瘤临床应用中的安全性和可行性。
