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Enhanced Effect of Mycobacterium tuberculosis-Related Antigens in Cervical Cancer Immunotherapy Based on Attenuated Listeria monocytogenes |
LIU Chen1, SONG Ya-Wen1, JIANG Xin1, SUN Jing1, JIANG Ling-Li2, SONG Hou-Hui1,*, CHENG Chang-Yong1,* |
1 College of Animal Science and Technology/College of Veterinary Medicine/Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics/Advanced Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management/China-Australia Joint Laboratory for Animal Health Big Data Analytics, Zhejiang A&F University, Hangzhou 311300, China; 2 NingBo College of Health Sciences, Ningbo 315100, China |
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Abstract Listeria monocytogenes has been increasingly studied as a delivery vector in various cancer and infectious disease immunotherapies, as it can induce robust immune responses in the host. In this study, the tumor vaccine strains, LM-Ag85B and LM-ESAT6 were constructed by fusing the Mycobacterium tuberculosis antigens Ag85B (Antigen 85B) and ESAT6 (6 kD early secreted antigen target) with the virulence factor listeriolysin O (LLO), which served as the basis in the preliminary laboratory experiments. The Western blot was employed to confirm the fusion expression and normal secretion of antigens with LLO. The in vitro growth capability and hemolytic activity of LM-Ag85B and LM-ESAT6 were investigated using bacterial infection biology methods. The therapeutic effects of combined application of LM-Ag85B and LM-ESAT6 with LM-E7 (expressing cervical cancer-related antigen E7) were evaluated in a mouse model of cervical cancer. The results revealed that LM-Ag85B and LM-ESAT6 were able to express and secrete the carried Ag85B and ESAT6 antigens, respectively. The in vitro growth capability of LM-Ag85B and LM-ESAT6 showed no significant difference compared to the wild-type strain, while their hemolytic activity was lower than that of the wild-type strain. In the mouse model of cervical cancer treatment, the experimental groups receiving combined application of LM-Ag85B, LM-ESAT6, and LM-E7 exhibited significant inhibition of tumor growth compared to the control group, indicating favorable therapeutic effects. This study demonstrates that the Mycobacterium tuberculosis antigens can provide enhanced tumor therapeutic efficacy for the LM-E7 based cervical cancer, which significantly promotes future applications of bacterial-based tumor immunotherapy..
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Received: 27 October 2022
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Corresponding Authors:
* Corresponding authors, songhh@zafu.edu.cn; lamge@zju.edu.cn
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[1] 陈璐, 汪枫婷, 孙静, 等. 2021. 减毒单增李斯特菌为载体的肿瘤免疫治疗研究进展[J]. 农业生物技术学报, 29(8): 1622-1629. (Chen L, Wang F T, Sun J, et al.2021. Research advances on attenuated Listeria monocytogenes as a vaccine platform for cancer immunotherapy[J]. Journal of Agricultural Biotechnology, 29(8): 1622-1629.) [2] 俞慧飞. 2020. 基于减毒单核细胞增多性李斯特菌为载体的肿瘤免疫技术探究[D]. 硕士学位论文, 浙江农林大学, 导师: 程昌勇. pp. 36-43. (Yu H Y.2020. A study on the attenuated Listeria monocytogenes-based tumor immunotherapy[D]. Thesis for M.S., Zhejiang Agriculture and Forestry University, Supervisor: Cheng C Y, pp. 36-43.) [3] Belisle J T, Vissa V D, Sievert T, et al.1997. Role of the major antigen of Mycobacterium tuberculosis in cell wall biogenesis[J]. Science, 276(5317): 1420-1422. [4] Buskwofie A, David-West G, Clare C A.2020. A review of cervical cancer: Incidence and disparities[J]. Journal of the National Medical Association, 112(2): 229-232. [5] Chen Z, Ozbun L, Chong N, et al.2014. Episomal expression of truncated listeriolysin O in LmddA-LLO-E7 vaccine enhances antitumor efficacy by preferentially inducing expansions of CD4+FoxP3- T cell and CD8+ T cells expansion[J]. Cancer Immunology Research, 2(9): 911-922. [6] Cheng C, Han X, Xu J, et al.2021. YjbH mediates the oxidative stress response and infection by regulating SpxA1 and the phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) in Listeria monocytogenes[J]. Gut Microbes, 13(1): 1-19. [7] Cheng C, Jiang L, Ma T, et al.2017. Carboxyl-terminal residues N478 and V479 required for the cytolytic activity of Listeriolysin o play a critical role in Listeria monocytogenes pathogenicity[J]. Frontiers in Immunology, 8: 1439. [8] Cheng C, Sun J, Yu H, et al.2020. Listeriolysin O pore-forming activity is required for ERK1/2 phosphorylation during Listeria monocytogenes infection[J]. Frontiers in Immunology, 11: 1146. [9] Cory L, Chu C.2014. ADXS-HPV: A therapeutic Listeria vaccination targeting cervical cancers expressing the HPV E7 antigen[J]. Human Vaccines and Immunotherapeutics, 10(11): 3190-3195. [10] Deng W, Lira V, Hudson T E, et al.2018. Recombinant Listeria promotes tumor rejection by CD8(+) T cell-dependent remodeling of the tumor microenvironment[J]. Proceedings of the National Academy of Sciencesof the USA, 115(32): 8179-8184. [11] Flickinger J C, Jr., Singh J, Yarman Y, et al.2022. T-cell responses to immunodominant Listeria epitopes limit vaccine-directed responses to the colorectal cancer antigen, guanylyl cyclase C[J]. Frontiers in Immunology, 13: 855759. [12] Ganguly N, Giang P H, Basu S K, et al.2007. Mycobacterium tuberculosis 6-kDa early secreted antigenic target (ESAT-6) protein downregulates lipopolysaccharide induced c-myc expression by modulating the extracellular signal regulated kinases 1/2[J]. BMC Immunology, 8: 24. [13] Gunn G R, Zubair A, Peters C, et al.2001. Two Listeria monocytogenes vaccine vectors that express different molecular forms of Human papilloma virus-16 (HPV-16) E7 induce qualitatively different T cell immunity that correlates with their ability to induce regression of established tumors immortalized by HPV-16[J]. The Journal of Immunology, 167(11): 6471-6479. [14] Guo L, Xie H, Zhang Z, et al.2021. Fusion protein vaccine based on Ag85B and STEAP1 induces a protective immune response against prostate cancer[J]. Vaccines (Basel), 9(7): 786. [15] Gu-Trantien C, Loi S, Garaud S, et al.2013. CD4+ follicular helper T cell infiltration predicts breast cancer survival[J]. Journal of Clinical Investigation, 123(7): 2873-2892. [16] Hadrup S, Donia M, Thor Straten P.2013. Effector CD4 and CD8 T cells and their role in the tumor microenvironment[J]. Cancer Microenviron, 6(2): 123-133. [17] Huh W K, Brady W E, Fracasso P M, et al.2020. Phase Ⅱstudy of axalimogene filolisbac (ADXS-HPV) for platinum-refractory cervical carcinoma: An NRG oncology/gynecologic oncology group study[J]. Gynecologic Oncology, 158(3): 562-569. [18] Karbalaei Zadeh Babaki M, Taghiabadi M, Soleimanpour S, et al.2019. Mycobacterium tuberculosis Ag85b:hfcγ1 recombinant fusion protein as a selective receptor-dependent delivery system for antigen presentation[J]. Microbial Pathogenesis, 129: 68-73. [19] Kumar R, Kumar P.2019. Yeast-based vaccines: New perspective in vaccine development and application[J]. FEMS Yeast Research, 19(2): 1-22 . [20] Kurnia I, Rauf S, Hatta M, et al.2022. Molecular Patho-mechanisms of cervical cancer (MMP1)[J]. Annals of Medicine and Surgery, 77: 103415. [21] Le D T, Dubensky T W, Brockstedt D G.2012. Clinical development of Listeria monocytogenes-based immunotherapies[J]. Seminars in Oncology, 39(3): 311-322. [22] Leclercq A, Charlier C, Lecuit M.2014. Global burden of listeriosis: The tip of the iceberg[J]. Lancet Infectious Diseases, 14(11): 1027-1028. [23] Liu Y, Lu Y, Ning B, et al.2022. Intravenous delivery of living Listeria monocytogenes elicits Gasdmermin-dependent tumor pyroptosis and motivates antitumor immune response[J]. ACS Nano, 16(3): 4102-4115. [24] Mignon C, Sodoyer R, Werle B.2015. Antibiotic-free selection in biotherapeutics: Now and forever[J]. Pathogens, 4(2): 157-181. [25] Moody C A, Laimins L A.2010. Human papillomavirus oncoproteins: Pathways to transformation[J]. Nature Reviews Cancer, 10(8): 550-560. [26] Oladejo M, Paterson Y, Wood L M.2021. Clinical experience and recent advances in the development of Listeria-based tumor immunotherapies[J]. Frontiers in Immunology, 12: 773. [27] Peng X, Treml J, Paterson Y.2007. Adjuvant properties of listeriolysin O protein in a DNA vaccination strategy[J]. Cancer Immunology Immunotherapy, 56(6): 797-806. [28] Shemesh C S, Hsu J C, Hosseini I, et al.2021. Personalized cancer vaccines: Clinical landscape, challenges, and opportunities[J]. Molecular Therapy, 29(2): 555-570. [29] Sung H, Ferlay J, Siegel R L, et al.2021. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA-A Cancer Journal for Clinicians, 71(3): 209-249. [30] Takeuchi A, Eto M, Tatsugami K, et al.2016. Antitumor activity of recombinant Bacille Calmette-Guérin secreting interleukin-15-Ag85B fusion protein against bladder cancer[J]. International Immunopharmacology, 35: 327-331. [31] Wang S, Guo J, Bai Y, et al.2022. Bacterial outer membrane vesicles as a candidate tumor vaccine platform[J]. Frontiers in Immunology, 13: 987419. [32] Yaghoubi A, Khazaei M, Hasanian S M, et al.2019. Bacteriotherapy in breast cancer[J]. International Journal of Molecular Sciences, 20(23): 5880. [33] Ylosmaki E, Manlio F, Beatriz M, et al.2021. Novel personalized cancer vaccine platform based on Bacillus Calmette-Guèrin 1 2[J]. Journal for ImmunoTherapy of Cancer, 9(7): e002707. [34] Zhao F, He X, Sun J, et al.2015. Cancer stem cell vaccine expressing ESAT-6-gpi and IL-21 inhibits melanoma growth and metastases[J]. American Journal of Translational Research, 7(10): 1870-1882. |
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WANG Feng-Ting, SUN Jing, LIU Chen, CHENG Yin, JIANG Xin, ZHU Yi-Ran, WEI Fang-Fang, ZHANG Xian, SONG Hou-Hui, CHENG Chang-Yong. Therapeutic Efficacy of Mouse (Mus musculus) Cervical Cancer Therapeutic Vaccine Based on the Attenuated Gene Deletion Listeria monocytogenes[J]. 农业生物技术学报, 2022, 30(10): 1987-1996. |
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