Therapeutic Effect of Recombinant Attenuated Listeria monocytogenes Expressing E7 Protein in a Mouse (Mus musculus) Model of Cervical Cancer
PAN Xiao-Xin, CUI Ming-Zhu, LUO Ya-Ru, SUN Jing, CHENG Chang-Yong, WANG Jing*, SONG Hou-Hui*
Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection & Internet Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine of Zhejiang A&F University, Hangzhou 311300, China
Abstract：Listeria monocytogenes is a high-quality vaccine candidate live vector that can migrate between cells and stimulate the body to promote the proliferation of antigen-specific CD4+ and CD8+ T cells to produce adaptive cellular immunity. This research group found that attenuated Listeria monocytogenes (LAM) had significantly lower hemolytic and proliferative ability than wild-type Listeria. At the same time, LAM could normally secrete Listeriolysin O (LLO) protein. In order to study the safety of LAM and its delivery and immune effect on the cervical cancer specific antigen E7 protein, this study tested the virulence of LAM in a mouse (Mus musculus) model, and to evaluate the role of attenuated Listeria monocytogenes-E7 (LAM-E7) in the immunotherapy and prevention of cervical cancer in mice. The results showed that the virulence of LAM to mice was 1/8709 of that of wild-type Listeria, and LAM could be eliminated from the organs of mice within a week. In this study, E7 gene was integrated into LAM genome by homologous recombination method. PCR identification results showed that LAM-E7 strain was successfully constructed, and Western blot results showed that E7 protein could be fused with LLO and expressed and released in the recombinant strain. The analysis of the growth ability and hemolytic activity of LAM-E7 in vitro showed that the fusion had no significant effect on the growth ability and hemolytic ability of LAM, indicating that the fusion of E7 protein would not affect the safety of LAM. In the cervical cancer treatment trial, the average tumor volume of the LAM-E7 group (161.1202 mm3) was notably smaller than that of the LAM group (349.4625 mm3) and the PBS group (512.4149 mm3)(P<0.01), indicating a significant inhibitory effected on tumor growth. The immunoprevention test showed that the average tumor volume of the LAM-E7 group (3.86 mm3) was significantly smaller than that of the LAM group (172.67 mm3) and the PBS group (57.38 mm3)(P<0.001), demonstrating the preventive effect of LAM-E7 on cervical cancer. LAM-E7 has demonstrated potent therapeutic and preventive properties on cervical cancer tumors in mice, which offers a new perspective for using attenuated Listeria as a vaccine vector in the treatment of cervical cancer. This study also provides valuable data that supports the advancement of therapeutic vaccines for cervical cancer in the future.
潘晓欣, 崔明珠, 罗亚如, 孙静, 程昌勇, 王晶, 宋厚辉. 表达E7蛋白的重组减毒李斯特菌在小鼠宫颈癌模型中的治疗效果[J]. 农业生物技术学报, 2024, 32(5): 1206-1215.
PAN Xiao-Xin, CUI Ming-Zhu, LUO Ya-Ru, SUN Jing, CHENG Chang-Yong, WANG Jing, SONG Hou-Hui. Therapeutic Effect of Recombinant Attenuated Listeria monocytogenes Expressing E7 Protein in a Mouse (Mus musculus) Model of Cervical Cancer. 农业生物技术学报, 2024, 32(5): 1206-1215.
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