Construction of Lmdd-LMP2A, an attenuated Listeria vaccine strain expressing the Epstein-Barr vi-rus latent membrane protein 2A (EBV-LMP2A) and evaluation of its anti-tumor effects against na-sopharyngeal carcinoma
10.3760/cma.j.issn.0254-5101.2015.03.011
- VernacularTitle:鼻咽癌特异性减毒重组疫苗 Lmdd-LMP2 A 的构建及其抑瘤效应的研究
- Author:
Wei ZHAO
;
Xiaoqin CHEN
;
Xin WAN
;
Ci CHENG
;
Zhe LIN
;
Dongju FENG
;
Kun YAO
;
Yun CHEN
- Publication Type:Journal Article
- Keywords:
Nasopharyngeal carcinoma;
Eepstein-barr virus;
Latent membrane protein 2A;
Liste-ria monocytogenes;
Tumor vaccine
- From:
Chinese Journal of Microbiology and Immunology
2015;(3):207-212
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare an attenuated Listeria vaccine Lmdd-LMP2A expressing the Ep-stein-Barr virus latent membrane protein 2A ( EBV-LMP2A) and evaluate its specific anti-tumor effects on nasopharyngeal carcinoma.Methods The gene fragment encoding EBV-LMP2A was amplified by PCR analysis and then subcloned into the shuttle vector p1565.PCR restriction enzyme digestion and DNA se-quencing were performed to identify the recombinant shuttle vector p1565-LMP2A.The p1565-LMP2A vector was then transformed into competent strains of the attenuated Listeria monocytogenes ( Lmdd) .The recombi-nant attenuated Listeria vaccine strain Lmdd-LMP2A was verified by Western blot assay.The histological sections of spleen and liver tissues were stained by Haematoxylin and eosin ( H&E) for analysis of inflamma-tion.A tumor-bearing HLA-A2 transgenic mouse model was established by subcutaneous injection of CNE-1/HLA-A2/LMP2A nasopharyngeal carcinoma cell line.The prepared Lmdd-LMP2A vaccine was inoculated into the mice via tail intravenous injection for the evaluation of specific CTL induction and the in vivo anti-tumor effects.Results The shuttle vector p1565-LMP2A and the recombinant attenuated Listeria vaccine strain Lmdd-LMP2A with stable expression of LMP2A protein were successfully constructed.The immunized mice showed mild inflammations with no structural damage and necrosis as indicated by H&E staining.The growth of tumors in tumor-bearing HLA-A2 transgenic mice was significantly inhibited by the immunization of Lmdd-LMP2A vaccine as compared with mice without inoculation.The survival time of mice was prolonged with the immunization of Lmdd-LMP2A vaccine.Conclusion The prepared attenuated Listeria vaccine Lm-dd-LMP2A showed specific anti-tumor effects with the safety advantage, suggesting the possibility of using it for anti-tumor therapy in clinic.
