孙冰洁;范鹏飞;房婷;于长明;陈薇;

• 1:军事科学院军事医学研究院生物工程研究所

摘要(Abstract)：

埃博拉病毒具有极大的危险性，对其活病毒相关的操作必须严格限制在生物安全四级实验室（BSL-4）中，阻碍了疫苗和治疗药物的研发进程。本研究将扎伊尔埃博拉病毒（EBOV）膜表面糖蛋白（GP）表达质粒与携带荧光素酶报告基因的pNL4-3.Luc.R-E-慢病毒载体共转染至HEK293T细胞，包装HIV骨架的EBOV GP-Fluc假病毒颗粒，建立了BSL-2条件下基于假病毒和生物发光成像（BLI）技术的BALB/c小鼠EBOV感染模型，并通过不同给药时间、途径和抗体种类的系统评估对其可靠性进行了验证。腹腔注射包含GP全长的假病毒在第4d可检测到集中于胸腺部位的最大发光信号，测试抗体在预防性静脉给药时可完全阻断假病毒的感染，在暴露后腹腔给药时表现出与其在活病毒小鼠模型中一致的保护效力。该模型为埃博拉病毒以及类似烈性病原体防治药物研发提供了一种简单有效的前期筛选方案。

关键词(KeyWords)： 埃博拉病毒;糖蛋白;假型病毒;小鼠模型;生物发光成像;抗体

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 孙冰洁;范鹏飞;房婷;于长明;陈薇;

DOI: 10.13242/j.cnki.bingduxuebao.004235

参考文献(References)：

• [1] Goldstein T, Anthony S J, Gbakima A, Bird B H,Bangura J, Tremeau-Bravard A, Belaganahalli M N,Wells H L, Dhanota J K, Liang E, Grodus M, Jangra R K, DeJesus V A, Lasso G, Smith B R, Jambai A,Kamara B O, Kamara S, Bangura W, Monagin C,Shapira S, Johnson C K, Saylors K, Rubin E M,Chandran K, Lipkin W I, Mazet J A K. The discovery of Bombali virus adds further support for bats as hosts of ebolaviruses[J]. Nat Microbiol, 2018, 3(10):1084-1089.
• [2] Hoenen T, Safronetz D, Groseth A, Wollenberg K R,Koita O A, Diarra B, Fall I S, Haidara F C, Diallo F,Sanogo M, Sarro Y S, Kone A, Togo A C, Traore A,Kodio M, Dosseh A, Rosenke K, de Wit E, Feldmann F, Ebihara H, Munster V J, Zoon K C, Feldmann H,Sow S. Virology. Mutation rate and genotype variation of Ebola virus from Mali case sequences[J]. Science,2015, 348(6230):117-119.
• [3] Dudas G, Carvalho L M, Bedford T, Tatem A J,Baele G, Faria N R, Park D J, Ladner J T, Arias A,Asogun D, Bielejec F, Caddy S L, Cotten M,D'Ambrozio J, Dellicour S, Di Caro A, Diclaro J W,Duraffour S, Elmore M J, Fakoli L S, Faye O, Gilbert M L, Gevao S M, Gire S, Gladden-Young A, Gnirke A, Goba A, Grant D S, Haagmans B L, Hiscox J A,Jah U, Kugelman J R, Liu D, Lu J, Malboeuf C M,Mate S, Matthews D A, Matranga C B, Meredith L W, Qu J, Quick J, Pas S D, Phan M V T, Pollakis G,Reusken C B, Sanchez-Lockhart M, Schaffner S F,Schieffelin J S, Sealfon R S, Simon-Loriere E, Smits S L, Stoecker K, Thorne L, Tobin E A, Vandi M A,Watson S J, West K, Whitmer S, Wiley M R,Winnicki S M, Wohl S, Wolfel R, Yozwiak N L,Andersen K G, Blyden S O, Bolay F, Carroll M W,Dahn B, Diallo B, Formenty P, Fraser C, Gao G F,Garry R F, Goodfellow I, Gunther S, Happi C T,Holmes E C, Kargbo B, Keita S, Kellam P, Koopmans M P G, Kuhn J H, Loman N J, Magassouba N,Naidoo D, Nichol S T, Nyenswah T, Palacios G,Pybus O G, Sabeti P C, Sall A, Stroher U, Wurie I,Suchard M A, Lemey P, Rambaut A. Virus genomes reveal factors that spread and sustained the Ebola epidemic[J]. Nature, 2017, 544(7650):309-315.
• [4] Chen R E, Zhang X, Case J B, Winkler E S, Liu Y,VanBlargan L A, Liu J, Errico J M, Xie X,Suryadevara N, Gilchuk P, Zost S J, Tahan S, Droit L, Turner J S, Kim W, Schmitz A J, Thapa M, Wang D, Boon A C M, Presti R M, O'Halloran J A, Kim A H J, Deepak P, Pinto D, Fremont D H, Crowe J E,Jr., Corti D, Virgin H W, Ellebedy A H, Shi P Y,Diamond M S. Resistance of SARS-CoV-2 variants to neutralization by monoclonal and serum-derived polyclonal antibodies[J]. Nat Med, 2021, 27(4):717-726.
• [5] Wang P, Nair M S, Liu L, Iketani S, Luo Y, Guo Y,Wang M, Yu J, Zhang B, Kwong P D, Graham B S,Mascola J R, Chang J Y, Yin M T, Sobieszczyk M,Kyratsous C A, Shapiro L, Sheng Z, Huang Y, Ho D D. Antibody resistance of SARS-CoV-2 variants B.1.351 and B.1.1.7[J]. Nature, 2021, 593(7857):130-135.
• [6] Kugelman J R, Kugelman-Tonos J, Ladner J T, Pettit J, Keeton C M, Nagle E R, Garcia K Y, Froude J W,Kuehne A I, Kuhn J H, Bavari S, Zeitlin L, Dye J M,Olinger G G, Sanchez-Lockhart M, Palacios G F.Emergence of Ebola virus escape variants in infected nonhuman primates treated with the MB-003 antibody cocktail[J]. Cell Rep, 2015, 12(12):2111-2120.
• [7] Li D, Chen T, Hu Y, Zhou Y, Liu Q, Zhou D, Jin X,Huang Z. An Ebola virus-like particle-based reporter system enables evaluation of antiviral drugs in vivo under non-biosafety level 4 conditions[J]. J Virol, 2016, 90(19):8720-8728.
• [8] Liu Q, Fan C, Li Q, Zhou S, Huang W, Wang L, Sun C, Wang M, Wu X, Ma J, Li B, Xie L, Wang Y.Antibody-dependent-cellular-cytotoxicity-inducing antibodies significantly affect the post-exposure treatment of Ebola virus infection[J]. Sci Rep, 2017, 7:45552.
• [9]李倩倩，埃博拉病毒等高致病性传染病假病毒小鼠感染模型的建立及应用. 2017，中国食品药品检定研究院.
• [10]Chen Q, Tang K, Zhang X, Chen P, Guo Y.Establishment of pseudovirus infection mouse models for in vivo pharmacodynamics evaluation of filovirus entry inhibitors[J]. Acta Pharm Sin B, 2018, 8(2):200-208.
• [11]Fan P, Chi X, Liu G, Zhang G, Chen Z, Liu Y, Fang T, Li J, Banadyga L, He S, Yu C, Qiu X, Chen W.Potent neutralizing monoclonal antibodies against Ebola virus isolated from vaccinated donors[J]. MAbs, 2020,12(1):1742457.
• [12]Holtsberg F W, Shulenin S, Vu H, Howell K A, Patel S J, Gunn B, Karim M, Lai J R, Frei J C, Nyakatura E K, Zeitlin L, Douglas R, Fusco M L, Froude J W,Saphire E O, Herbert A S, Wirchnianski A S, LearRooney C M, Alter G, Dye J M, Glass P J, Warfield K L, Aman M J. Pan-ebolavirus and pan-filovirus mouse monoclonal antibodies:protection against Ebola and Sudan viruses[J]. J Virol, 2016, 90(1):266-278.
• [13]Qiu X, Fernando L, Melito P L, Audet J, Feldmann H, Kobinger G, Alimonti J B, Jones S M. Ebola GPspecific monoclonal antibodies protect mice and guinea pigs from lethal Ebola virus infection[J]. PLoS Negl Trop Dis, 2012, 6(3):e1575.
• [14]Zhao X, Howell K A, He S, Brannan J M, Wec A Z,Davidson E, Turner H L, Chiang C I, Lei L, Fels J M, Vu H, Shulenin S, Turonis A N, Kuehne A I, Liu G, Ta M, Wang Y, Sundling C, Xiao Y, Spence J S,Doranz B J, Holtsberg F W, Ward A B, Chandran K,Dye J M, Qiu X, Li Y, Aman M J. Immunizationelicited broadly protective antibody reveals Ebolavirus fusion loop as a site of vulnerability[J]. Cell, 2017, 169(5):891-904 e815.
• [15]Rhein B A, Maury W J. Ebola virus entry into host cells:identifying therapeutic strategies[J]. Curr Clin Microbiol Rep, 2015, 2(3):115-124.
• [16]Gunn B M, Yu W H, Karim M M, Brannan J M,Herbert A S, Wec A Z, Halfmann P J, Fusco M L,Schendel S L, Gangavarapu K, Krause T, Qiu X, He S, Das J, Suscovich T J, Lai J, Chandran K, Zeitlin L, Crowe J E, Jr., Lauffenburger D, Kawaoka Y,Kobinger G P, Andersen K G, Dye J M, Saphire E O,Alter G. A role for Fc function in therapeutic monoclonal antibody-mediated protection against Ebola virus[J].Cell Host Microbe, 2018, 24(2):221-233 e225.
• [17]Bournazos S. IgG Fc receptors:evolutionary considerations[J]. Curr Top Microbiol Immunol, 2019,423:1-11.
• [18]Saphire E O, Schendel S L, Fusco M L, Gangavarapu K, Gunn B M, Wec A Z, Halfmann P J, Brannan J M,Herbert A S, Qiu X, Wagh K, He S, Giorgi E E,Theiler J, Pommert K B J, Krause T B, Turner H L,Murin C D, Pallesen J, Davidson E, Ahmed R, Aman M J, Bukreyev A, Burton D R, Crowe J E, Jr., Davis C W, Georgiou G, Krammer F, Kyratsous C A, Lai J R, Nykiforuk C, Pauly M H, Rijal P, Takada A,Townsend A R, Volchkov V, Walker L M, Wang C I,Zeitlin L, Doranz B J, Ward A B, Korber B, Kobinger G P, Andersen K G, Kawaoka Y, Alter G, Chandran K, Dye J M, Viral Hemorrhagic Fever Immunotherapeutic C. Systematic analysis of monoclonal antibodies against Ebola virus GP defines features that contribute to protection[J]. Cell, 2018, 174(4):938-952 e913.