Effects of the number of amino acid residues in the signal segment upstream or downstream of the NS2B-3 cleavage site on production and secretion of prM/M-E virus-like particles of West Nile virus |
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| Authors: | Hidehiro Takahashi Naohiro Ohtaki Masae Maeda-Sato Michiko Tanaka Keiko Tanaka Hirofumi Sawa Toyokazu Ishikawa Akihisa Takamizawa Tomohiko Takasaki Hideki Hasegawa Tetsutaro Sata William W. Hall Takeshi Kurata Asato Kojima |
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| Affiliation: | 1. Department of Pathology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan;2. Department of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan;3. Global COE Program, Hokkaido University, Sapporo 001-0020, Japan;4. Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Yahata-cho, Kanonji-shi, Kagawa 768-0061, Japan;5. Department of Virology I, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, Japan;6. Department of Medical Microbiology, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland;1. The Urogenital Sub-committee and the Surveillance Committee of Japanese Society of Chemotherapy (JSC), The Japanese Association for Infectious Diseases (JAID) and The Japanese Society for Clinical Microbiology (JSCM), Tokyo, Japan;2. The Surveillance Committee of JSC, JAID and JSCM, Tokyo, Japan;3. Department of Urology, University of Occupational and Environmental Health, Kitakyushu, Japan;4. Department of Urology, Gifu University Hospital, Gifu, Japan;5. Department of Urology, School of Medicine, Fujita Health University, Toyoake, Japan;6. Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan;7. Blood Purification Center, Kagoshima University Hospital, Kagoshima, Japan;8. Department of Urology, Sapporo Medical University School of Medicine, Sapporo, Japan;9. Department of Urology, Hyogo College of Medicine, Nishinomiya, Japan;10. Research Center for Anti-infectious Drugs, Kitasato Institute for Life Sciences, Kitasato University, Tokyo, Japan;11. Department of Urology, The Jikei University Katsushika Medical Center, Tokyo, Japan;12. Department of Urology, The Jikei University School of Medicine, Tokyo, Japan;13. Division of Urology, Kobe University Graduate School of Medicine, Kobe, Japan;14. Department of Urology, Institute of Biomedical & Health Sciences Hiroshima University, Hiroshima, Japan;15. Department of Urology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan;p. Hirayama Urology Clinic, Kumamoto, Japan;q. Narita Clinic, Nagoya, Japan;r. Hosobe Clinic, Tokyo, Japan;s. iClinic, Sendai, Japan;t. Ito Urology Clinic, Kitakyushu, Japan;u. Kawai Urology Clinic, Kitakyushu, Japan;v. Gifu Urological Clinic, Gifu, Japan;w. Chokyu Tenma Clinic, Himeji, Japan;x. Matsumura Urology Clinic, Kato, Japan;y. Yoshioka Urology Clinic, Nishinomiya, Japan;z. Hirajima Clinic, Okayama, Japan;11. Araki Urological Clinic, Kurashiki, Japan;12. Department of Urology, Takayama Hospital, Chikushino, Japan;13. Kaji Clinic, Fukuoka, Japan;14. Kawahara Urology Clinic, Kagoshima, Japan;15. Sumii Clinic, Hiroshima, Japan;16. Kadena Urological Clinic, Hiroshima, Japan;17. Yamaguchi Dermatology and Urology Clinic, Munakata, Japan;18. Department of Urology, Toyota Memorial Hospital, Toyota, Japan;19. Nishi Urology and Dermatology Clinic, Kitakyushu, Japan;110. Nishimura Urology Clinic, Kitakyushu, Japan;111. Shirane Urology Clinic, Aki-gun, Japan;112. Yoh Urology and Dermatology Clinic, Inazawa, Japan;113. Akiyama Urology Clinic, Nishinomiya, Japan;114. Imai Urology Clinic, Akashi, Japan;115. Department of Urology, Kano Hospital, Kasuya-gun, Japan;1. Unit of PharmacoEpidemiology & PharmacoEconomics, Department of Pharmacy, University of Groningen, The Netherlands;2. Global Market Access Solutions (GMAS), St-Prex, Switzerland;3. Initiative for Vaccine Research, World Health Organization, Geneva, Switzerland;1. Faculty of Medicine, Department of Chemistry, University of Niš, Niš, Serbia;2. IRCCS- Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy;1. State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;2. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, Hangzhou 310003, Zhejiang, China |
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| Abstract: | Expression of genes for precursor M (prM) and envelope (E) proteins of West Nile virus (WNV) leads to the production of small, capsidless, and non-infectious virus-like particles (VLPs) possessing the E antigen which is responsible for viral entry and immune protection. It has been reported that processing of the secretion signal affects viral release. We examined the secretion efficiency of VLPs into the culture medium from RK13 or 293 T cells transfected with expression vectors for prM and E proteins of WNV which were constructed to comprise different lengths of signal peptides upstream of the prM-E domain. The number of amino acid residues present in the segment markedly affected the production, processing, and secretion of VLPs. Secreted VLPs possessed both the processed M protein and the glycosylated E protein. In addition, immunization with VLPs induced neutralizing antibodies in C3H/HeN mice. These results indicate that the number of amino acid residues comprising the N-terminus of the signal segment controls the efficiency of assembly, maturation, and release of VLPs in the absence of viral protease, which in turn indicates the potential of VLPs as a candidate for an effective WNV subunit vaccine. |
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| Keywords: | West Nile virus prM VLP |
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