Baculovirus surface display of the HA protein of H5N2 avian influenza virus and its immunogenicity against a lethal challenge with H5N1 virus in chickens

被引:11
|
作者
Tung, Min-Che [1 ]
Lu, Hsin-Yu [2 ]
Chang, Yu-Kang [1 ,11 ]
Huang, Wei-Ru [2 ]
Liao, Tsai-Ling [3 ,4 ,5 ]
Wu, Hung-Yi [6 ]
Chang, Ching-Dong [6 ]
Fan, Hueng-Chuen [1 ]
Nielsen, Brent L. [7 ]
Liu, Hung-Jen [2 ,4 ,5 ,8 ,9 ,10 ]
机构
[1] Tungs Taichung Metroharbor Hosp, Dept Stomatol & Med Res, Taichung, Taiwan
[2] Natl Chung Hsing Univ, Inst Mol Biol, Taichung 402, Taiwan
[3] Taichung Vet Gen Hosp, Dept Med Res, Taichung 407, Taiwan
[4] Natl Chung Hsing Univ, Rong Hsing Res Ctr Translat Med, Taichung 402, Taiwan
[5] Natl Chung Hsing Univ, PhD Program Translat Med, Taichung 402, Taiwan
[6] Natl Pingtung Univ Sci & Technol, Dept Vet Med, Pingtung 912, Taiwan
[7] Brigham Young Univ, Dept Microbiol & Mol Biol, Provo, UT 84602 USA
[8] Natl Chung Hsing Univ, iEGG, Taichung 402, Taiwan
[9] Natl Chung Hsing Univ, Anim Biotechnol Ctr, Taichung 402, Taiwan
[10] Natl Chung Hsing Univ, Dept Life Sci, Taichung, Taiwan
[11] Jen Teh Jr Coll Med & Management, Dept Nursing, Hou Loung Town, Taiwan
关键词
Avian influenza virus; HA protein; Baculovirus gp64 cytoplasmic domain; Baculovirus surface display system; Multiple expression cassettes; VACCINATED ANIMALS STRATEGY; REVERSE GENETICS; MAMMALIAN-CELLS; EXPRESSION; PROTECTION; INFECTION; ENVELOPE;
D O I
10.1016/j.vetmic.2020.108640
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
In the present study, we have generated several H5N2 HA recombinant baculoviruses for production of a HA subunit vaccine against the lethal H5N2 avian influenza virus (AIV). The effective display of functional HA on the cell membrane and baculoviral envelope was examined. Our results reveal that chickens immunized with the chimeric AIV HA protein fused with the baculovirus gp64 cytoplasmic domain (CTD) induced higher HI titer. To further increase the expression level of the H5N2 AIV HA protein, the HA gene of H5N2 AIV was amplified and cloned into three novel baculovirus surface display vectors BacDual DisplayEGFP-2HA, BacDual DisplayEGFP-3HA, BacDual DisplayEGFP-4HA which contains multiple expression cassettes for higher level display of HA proteins on the cell membrane and baculovirus envelope. To determine the optimum conditions for producing HA protein, various MOI, infection times, and shaker times for virus transfection were tested. Our results reveal that the conditions of an MOI of 5, 3 day post infection, and 15 min of shaker time have higher efficiency for HA protein production. Our results reveal that the baculovirus surface display vector pBacDual DisplayEGFP-4HA increases significantly the expression level of the H5N2 AIV HA protein. Chickens that received two doses of BacDual DisplayEGFP-4HA cell lysates formulated with Montanide ISA70 adjuvant elicited efficient immunogenicity and had an average HI titer of 7 log 2 at 2 weeks post-vaccination. Challenge studies revealed that vaccinated chickens with HI titers 5 log 2 were completely protected against the lethal H5N1 AIV challenge. Furthermore, HI titers could be maintained at 5 log 2 for 20 weeks for laying hens. This study suggests that the HA protein expression from the baculovirus surface display system could be a safe and efficacious subunit vaccine for chickens.
引用
收藏
页数:10
相关论文
共 50 条
  • [31] Avian Influenza A (H5N1) Virus and 2 Fundamental Questions
    Briand, Sylvie
    Fukuda, Keiji
    JOURNAL OF INFECTIOUS DISEASES, 2009, 199 (12): : 1717 - 1719
  • [32] Vaccination against Human Influenza A/H3N2 Virus Prevents the Induction of Heterosubtypic Immunity against Lethal Infection with Avian Influenza A/H5N1 Virus
    Bodewes, Rogier
    Kreijtz, Joost H. C. M.
    Baas, Chantal
    Geelhoed-Mieras, Martina M.
    de Mutsert, Gerrie
    van Amerongen, Geert
    van den Brand, Judith M. A.
    Fouchier, Ron A. M.
    Osterhaus, Albert D. M. E.
    Rimmelzwaan, Guus F.
    PLOS ONE, 2009, 4 (05):
  • [33] Vaccination of chickens against H5N1 avian influenza in the face of an outbreak interrupts virus transmission
    Ellis, TM
    Leung, CYHC
    Chow, MKW
    Bissett, LA
    Wong, W
    Guan, Y
    Peiris, JSM
    AVIAN PATHOLOGY, 2004, 33 (04) : 405 - 412
  • [34] Development of DNA Vaccine against Avian Influenza A Virus (H5N1)
    Ahmed, Shaheen
    Bronich, Tatiana K.
    Kabanov, Alexander V.
    MOLECULAR THERAPY, 2011, 19 (07) : 1392 - 1393
  • [35] Role of different genes in the pathogenesis of H5N1 avian influenza virus in chickens
    Wasilenko, J. L.
    Lee, C. W.
    Sarmento, L.
    Spackman, E.
    Suarez, D. L.
    Pantin-Jackwood, M. J.
    POULTRY SCIENCE, 2008, 87 : 156 - 156
  • [36] Avian influenza virus (H5N1) mortality surveillance
    Komar, Nicholas
    Olsen, Bjorn
    EMERGING INFECTIOUS DISEASES, 2008, 14 (07) : 1176 - 1178
  • [38] Detection of avian influenza a H5N1 virus by pyrosequencing
    Wu, Wenjuan
    Zhou, Guohua
    Progress on Post-Genome Technologies, 2006, : 103 - 106
  • [39] Influenza viruses and the evolution of avian influenza virus H5N1
    Skeik, Nedaa
    Jabr, Fadi I.
    INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES, 2008, 12 (03) : 233 - 238
  • [40] Phylodynamics of H5N1 avian influenza virus in Indonesia
    Lam, Tommy Tsan-Yuk
    Hon, Chung-Chau
    Lemey, Philippe
    Pybus, Oliver G.
    Shi, Mang
    Tun, Hein Min
    Li, Jun
    Jiang, Jingwei
    Holmes, Edward C.
    Leung, Frederick Chi-Ching
    MOLECULAR ECOLOGY, 2012, 21 (12) : 3062 - 3077