Construction and characterization of attenuated recombinant Listeria monocytogenes delivering HPV16 E7

Yanyan Jia1 Yuelan Yin1 Weijun Tan1 Feifei Duan1 Zhiming Pan1 Xiang Chen1 Xin′an Jiao1

(1.Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, Jiangsu, China 225009)

【Abstract】 Listeria monocytogenes (LM) is an excellent tumor vaccine vector. In this study, recombinant LM expressing human papillomavirus type 16 (HPV16) E7 protein, a vaccine candidate was constructed and the biological characteristics were determined. Through homologous recombination, E7 gene was cloned in frame with the LM4 hly promoter-signal sequence, and introduced into the LM4 genome. The recombinants, constructed as LM4△hly::E7, were plasmid- and antibiotic-resistant gene-free. LM4△hly::E7 could express and secrete a 66 kDa E7-LLO fusion protein, which had immunological activities. Proliferation of LM4△hly::E7 in RAW264.7 macrophages was observed under confocal laser scanning microscope. Specific anti-E7 antibodies could be detected by ELISA in mice immunized with LM4△hly::E7. The LD50 of LM4△hly::E7 was 3.863 × 109 CFU in C57BL/6 mice with intraperitoneal immunization, which was 104 times higher than that of wild type LM4. No obvious pathological changes were detected in the immunized mice. These results suggest that LM4△hly::E7 expressing E7-LLO fusion protein may be a safe and promising vector-vaccine candidate for cervical cancer, and may also be important biological materials for anti-tumor research.

【Keywords】 attenuated Listeria monocytogenes; HPV16 E7; macrophage cell line; safety;


【Funds】 National Natural Science Foundation of China (Nos. 31472193, 31101841) National Natural Science Foundation of China(Nos.31472193,31101841) Priority Academic Program Development of Jiangsu Higher Education Institutions Priority Academic Program Development of Jiangsu Higher Education Institutions

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(Translated by Chengji)


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This Article


CN: 11-1998/Q

Vol 32, No. 05, Pages 683-692

May 2016


Article Outline



  • 1 Materials and methods
  • 2 Results and analyses
  • 3 Discussion
  • References