Biểu hiện Protein HA/H7N9 Polymer dung hợp IgMFc trong cây thuốc lá (Nicotiana benthamiana) bằng phương pháp Agroinfiltration
Main Article Content
Abstract
Virus cúm A/H7N9 gây bệnh trên người xuất hiện đầu tiên tại Trung Quốc năm 2013 và tiếp tục lây nhiễm đến nay với tỷ lệ tử vong 40%. Hemagglutinin (HA) là protein chính của vỏ virus cúm, chứa các epitope trung hòa virus, được xem như là mục tiêu hàng đầu dùng để thiết kế loại vắc xin tái tổ hợp chống lại sự xâm nhiễm của virus cúm A. Trong nghiên cứu này, kháng nguyên HA của virus cúm H7N9 dung hợp với đoạn Fc của IgM được biểu hiện tạm thời trên cây thuốc lá nicotiana. sp bằng phương pháp Agro-infiltration. Sự biểu hiện của protein được kiểm tra bằng lai miên dịch. Kết quả cho thấy, chúng tôi đã biểu hiện thành công protein (H7pII-IgMFc)3 trong cây thuốc lá N. benthamiana. Protein tái tổ hợp được tinh sạch bằng phương pháp sắc ký ion cố định kim loại (Immobilized metal ion chromatography- IMAC) và đánh giá hoạt tính sinh học bằng phản ứng ngưng kết hồng cầu. Kết quả kiểm tra hoạt tính cho thấy (H7pII-IgMFc)3 gây ngưng kết hồng cầu với hiệu giá ngưng kết 32 HAU tương ứng với 0,31 µg protein tinh sạch. Kết quả này mở ra một hướng mới cho việc phát triển vắc xin chống virus cúm A/H7N9 trong tương lai.
Keywords:
Virus cúm A/H7N9, Hemagglutinin (HA), polymer dung hợp IgMFc, biểu hiện tạm thời, protein tái tổ hợp.
References
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[2] www.who.int
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[4] Min JY, Vogel L, Matsuoka Y, Lu B, Swayne D, Jin H, et al, A live attenuated H7N7 candidate vaccine virus induces neutralizing antibody that confers protection from challenge in mice, ferrets, and monkeys, J Virol 84(22) (1010)11950–60. [PubMed: 20810733].
[5] Couch RB, Patel SM, Wade-Bowers CL, Nino D, A randomized clinical trial of an inactivated avian influenza A (H7N7) vaccine, PLoS ONE 7(12) (2012) e49704. [PubMed: 23239968].
[6] Cox RJ, Major D, Hauge S, Madhun AS, Brokstad KA, Kuhne M, A cell-based H7N1 split influenza virion vaccine confers protection in mouse and ferret challenge models, Influenza Other Respir Viruses 3(3) (2009) 107–17. [PubMed: 19453487].
[7] Goff PH, Krammer F, Hai R, Seibert CW, Margine I, Garcia-Sastre A, Induction of cross-reactive antibodies to novel H7N9 influenza virus by recombinant Newcastle disease virus expressing a North American lineage H7 subtype hemagglutinin, J Virol 87(14) (2013) 8235–40. [PubMed: 23698299].
[8] Kreijtz JH, Wiersma LC, De Gruyter HL, Vogelzang-van Trierum SE, van Amerongen G, Stittelaar KJ, A single immunization with modified vaccinia virus ankara-based influenza virus H7 vaccine affords protection in the influenza A(H7N9) pneumonia ferret model, J Infect Dis. (2014).
[9] Gaspar LP, Mendes YS, Yamamura AMY, Almeida LFC, Caride E, Goncalves RB, Pressure-inac-tivated yellow fever 17DD virus: Implications for vaccine development, J Virol Methods 150 (2008) 57–62. doi: 10.1016/ j.jviromet.2008.03.002 PMID: 18420285.
[10] Galarza JM, Latham T, Cupo A, Virus-like particle (VLP) vaccine conferred complete protection against a lethal influenza virus challenge, Viral Immunol 18(1) (2005) 244–51. [PubMed: 15802970].
[11] Krammer F, Albrecht RA, Tan GS, Margine I, Hai R, Schmolke M, Divergent H7 immunogens offer protection from H7N9 virus challenge, J Virol 88(8) (2014) 3976–85. [PubMed: 24453375].
[12] Kang SM, Pushko P, Bright RA, Smith G, Compans RW, Influenza virus-like particles as pandemic vaccines, Curr Top Microbiol Immunol 333 (2009) 269–89. [PubMed: 19768411].
[13] Pushko P, Pujanauski L.M, Sun X, Pearce M, Hidajat R, KortT, Schwartzman L.M,TretyakovaI, Chunqing L, Taubenberger J.K, Tumpey T.M, Recombinant H7 hemagglutinin forms subviral particles that protect mice and ferrets from challenge with H7N9 influenza virus, Vaccine 33(38) (2015) 4975–4982. doi:10.1016/j.vaccine. 2015.07.026.
[14] Pushko P, Pumpens P, Grens E, Development of virus-like particle technology from small highly symmetric to large complex virus-like particle structures, Intervirology 56(3) (2013) 141–65. [PubMed: 23594863].
[15] Boes M, Role of natural and immune IgM antibodies in immune responses, Mol Immunol 37(18) (200), 1141–1149.
[16] Andrianov V., R. Brodzik, S. Spitsin, Production of recombinant anthrax toxin receptor (ATR/CMG2) fused with human Fcin planta, Protein Expression and Purification, 70 (2) (2010) 158–162.
[17] Fischer, R., Schumann,D., Zimmermann,S., Drossard,J., Sack,M., and Schillberg,S., Expression and characterization of bispecific single-chain Fv fragments produced in transgenic plants, European Journal of Biochemistry 262(1999) 810-816.
[18] Shoji, Y.; Bi, H.; Musiychuk, K.; Rhee, A.; Horsey, A.; Roy, G.; Green, B.; Shamloul, M.; Farrance, C.E., Taggart, B., Plant-derived hemagglutinin protects ferrets against challenge infection with the A/Indonesia/05/05 strain of avian influenza, Vaccine (27) (2009) 1087–1092.
[19] Mortimer E, Maclean J.M,Mbewana S, Buys A,Williamson A.L, Hitzero I.I, Rybicki E.P, Setting up a platform for plant-based influenza virus vaccine production in South Africa, BMC Biotechnology 12 (2012)14. DOI: 10.1186/ 1472-6750-12-14.
[20] Mett, V., Musiychuk, K., Bi, H., Farrance, C.E., Horsey, A., Ugulava, N., Shoji, Y., de la Rosa, P., Palmer, G.A., Rabindran, S., Streatfield, S.J., Boyers, A., Russell, M., Mann, A., Lambkin, R., Oxford, J.S., Schild, G.C., and Yusibov, V., A plant-produced influenza subunit vaccine protects ferrets against virus challenge, Influenza Other Respi Viruses 2 (2008), 33-40.
[21] Xiang C, Han P, Lutziger I, Wang K, Oliver DJ, A mini binary vector series for plant transformation, Plant Mol Biol 40 (1999), 711–717.
[22] Deblaere R., Bytebier B., De Greve, H., Deboeck F., Schell J., Van Montagu M., and Leemans J., Efficient octopine Ti plasmidderived vectors for Agrobacterium-mediated gene transfer to plants, Nucleic Acids Res. 13 (1985) 4777-4788.
[23] http://www.oic.gov.ie/en/Publications/Annual-Reports/2014-Annual-Report.
[24] Phan, H.T., Hause, B., Hause, G., Arcalis, E., Stoger, E., Maresch, D., Altmann, F., Joensuu, J., Conrad, U., Influence of Elastin-Like Polypeptide and hydrophobin on recombinant Hemagglutinin accumulations in transgenic tobacco plants, PLoS ONE 9(6) (2014) e99347.
[25] Krishnamurthy Konduru, Steven B. Bradfute, Jerome Jacques, Mohanraj Manangeeswaran, Ebola virus glycoprotein Fc fusion protein confers protection against lethal challenge in vaccinated mice, Vaccine 29(16) (2011) 2968–2977.
[26] David N. A. Mekhaiel, Daniel M. Czajkowsky, Jan Terje Andersen, Jianguo Shi, Marwa El-Faham,Polymeric human Fc-fusion proteins with modified effector functions, SCIENTIFICREPO RTS(2011) 1: 124. DOI: 10.1038/srep00124.
[27] Keith L. Wycoff, Archana Belle, Dorothe´e Deppe, Leah Schaefer, James M. Maclean, Recombinant Anthrax Toxin Receptor-Fc Fusion Proteins Produced in Plants Protect Rabbits against Inhalational Anthrax, ANTIMICROBIA LAGENTS ANDCHEMOTHERAPY 55(2011) 132–139.
[28] Loureiro Silvia, Junyuan Ren, Pongsathon Phapugrangkul, Camilo A. Colaco, Christopher R., Adjuvant-Free Immunization with Hemagg lutinin-Fc Fusion Proteins as an Approach to Influenza Vaccines, JOURNAL OF VIROLOGY (2011) 3010–3014.