Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b <i>in vitro</i>

Danrong Shi; Keda Chen; Xiangyun Lu; Linfang Cheng; Tianhao Weng; Fumin Liu; Nanping Wu; Lanjuan Li; Hangping Yao

doi:10.1016/j.virs.2022.01.031

April 2022

Danrong Shi, Keda Chen, Xiangyun Lu, Linfang Cheng, Tianhao Weng, Fumin Liu, Nanping Wu, Lanjuan Li and Hangping Yao. Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro[J]. Virologica Sinica, 2022, 37(2): 295-298. doi: 10.1016/j.virs.2022.01.031

Citation: Danrong Shi, Keda Chen, Xiangyun Lu, Linfang Cheng, Tianhao Weng, Fumin Liu, Nanping Wu, Lanjuan Li, Hangping Yao. Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro .VIROLOGICA SINICA, 2022, 37(2) : 295-298. http://dx.doi.org/10.1016/j.virs.2022.01.031

重组人干扰素α1b对新型冠状病毒的抑制作用优于α2b

史丹蓉 ^a ,
陈科达 ^b ,
卢翔云 ^a ,
程林芳 ^a ,
翁天豪 ^a ,
刘福民 ^a ,
吴南屏 ^a,, ,
李兰娟 ^a,, ,
姚航平 ^a,,

a 浙江大学医学院附属第一医院传染病诊治国家重点实验室、国家感染性疾病临床医学研究中心、国家传染病医学中心, 杭州 310003
b 浙江树人大学树兰国际医学院, 杭州 310015

通讯作者： 吴南屏, flwnp2013@163.com; 李兰娟, ljli@zju.edu.cn; 姚航平, yaohangping@zju.edu.cn
收稿日期： 2021-02-20
录用日期： 2022-01-21

摘要

在新型冠状病毒肺炎疫情的蔓延下，我们迫切需要寻找一种有效抗病毒药物来遏制疫情。迄今为止，对新冠肺炎的治疗还缺乏十分有效的药物。本文报道了重组人干扰素α1b对新冠病毒的体外抑制作用。本研究在细胞模型非洲绿猴肾细胞Vero和人肺腺癌细胞Calu-3上用CCK-8法检测重组人干扰素α1b和α2b的剂量毒性效应，进而采用逆转录定量聚合酶链反应（RT-qPCR）、半数组织培养感染剂量（TCID50）法和免疫荧光等方法评估重组人干扰素α1b和α2b的抗新冠病毒活性。结果显示，重组人干扰素α1b在检测浓度范围内对Vero细胞和Calu-3细胞均没有细胞毒性（CC50>25000 IU/mL），且在这两种细胞模型中，均表现出显著的抗新冠病毒作用（在Vero细胞中：EC₅₀=0.12 IU/mL；在Calu-3细胞中：EC₅₀=0.52 IU/mL），优于重组人干扰素α2b（在Vero细胞中：EC₅₀=0.25 IU/mL；在Calu-3细胞中：EC₅₀=2.48 IU/mL）。本研究显示重组人干扰素α1b对新型冠状病毒具有良好的体外抑制作用，是临床治疗新冠肺炎的潜在药物。
- 新型冠状病毒
- , 抗病毒药物
- , 重组人干扰素α1b
- , 重组人干扰素α2b
- , 瑞德西韦

Recombinant human interferon-α1b inhibits SARS-CoV-2 better than interferon-α2b in vitro

Danrong Shi ^a ,
Keda Chen ^b ,
Xiangyun Lu ^a ,
Linfang Cheng ^a ,
Tianhao Weng ^a ,
Fumin Liu ^a ,
Nanping Wu ^a,, ,
Lanjuan Li ^a,, ,
Hangping Yao ^a,,

a State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China
b Shulan International Medical College, Zhejiang Shuren University, Hangzhou, 310015, China

Corresponding author: Nanping Wu, flwnp2013@163.com Lanjuan Li, ljli@zju.edu.cn Hangping Yao, yaohangping@zju.edu.cn
Received Date: 20 February 2021
Accepted Date: 21 January 2022

Abstract

Highlights
1) A comprehensive evaluation method for anti-SARS-CoV-2 drugs was established based on RT-qPCR, TCID₅₀ method, and immunofluorescence.
2) A significant antiviral effect of rHuIFN-α1b was shown with EC₅₀=0.12 IU/mL in Vero cells and EC₅₀=0.52 IU/mL in Calu-3 cells, which was better than rHuIFN-α2b (EC₅₀=0.25 IU/mL in Vero cells and EC₅₀=2.48 IU/mL in Calu-3 cells).
3) rHuIFN-α1b has a good potential in the application of anti-COVID-19 therapy.

References
1. Ader, F., Peiffer-Smadja, N., Poissy, J., Bouscambert-Duchamp, M., Belhadi, D., Diallo, A., Delmas, C., Saillard, J., Dechanet, A., Mercier, N., Dupont, A., Alfaiate, T., Lescure, F.X., Raffi, F., Goehringer, F., Kimmoun, A., Jaureguiberry, S., Reignier, J., Nseir, S., Danion, F., Clere-Jehl, R., Bouiller, K., Navellou, J.C., Tolsma, V., Cabié, A., Dubost, C., Courjon, J., Leroy, S., Mootien, J., Gaci, R., Mourvillier, B., Faure, E., Pourcher, V., Gallien, S., Launay, O., Lacombe, K., Lanoix, J.P., Makinson, A., MartinBlondel, G., Bouadma, L., Botelho-Nevers, E., Gagneux-Brunon, A., Epaulard, O., Piroth, L., Wallet, F., Richard, J.C., Reuter, J., Staub, T., Lina, B., Noret, M., Andrejak, C., Lê, M.P., Peytavin, G., Hites, M., Costagliola, D., Yazdanpanah, Y., Burdet, C., Mentré, F., DisCoVeRy study group, 2021. An open-label randomized, controlled trial of the effect of lopinavir/ritonavir, lopinavir/ritonavir plus IFN-β-1a and hydroxychloroquine in hospitalized patients with COVID-19. Clin. Microbiol. Infect. 27, 1826–1837.
2. Al-Badr, A.A., Ajarim, T.D.S., 2018. Ganciclovir. Profiles Drug Subst. Excip. Relat. Methodol. 43, 1–208.
3. Cao, Y.C., Deng, Q.X., Dai, S.X., 2020. Remdesivir for severe acute respiratory syndrome coronavirus 2 causing COVID-19: an evaluation of the evidence. Trav. Med. Infect. Dis. 35, 101647.
4. Centers for Disease Control and Prevention (CDC), 2021. COVID-19: SARS-CoV-2 Variant Classifications and Definitions. US Department of Health and Human Services, CDC, Atlanta, GA. Available: https://www.cdc.gov/coronavirus/2019-ncov/cases-updates/variant-surveillance/variant-info.html. (Accessed 1 December 2021).
5. Chen, L., Shi, M., Deng, Q., Liu, W., Li, Q., Ye, P., Yu, X., Zhang, B., Xu, Y., Li, X., Yang, Y., Li, M., Yan, Y., Xu, Z., Yu, J., Xiang, L., Tang, X., Wan, G., Cai, Q., Wang, L., Hu, B., Xie, L., Li, G., Xie, L., Liu, X., Liu, C., Li, L., Chen, L., Jiang, X., Huang, Y., Wang, S., Guo, J., Shi, Y., Li, L., Wang, X., Zhao, Z., Li, Y., Liu, Y., Fu, Q., Zeng, Y., Zou, Y., Liu, D., Wan, D., Ai, T., Liu, H., 2020. A multi-center randomized prospective study on the treatment of infant bronchiolitis with interferon alpha1b nebulization. PLoS One 15, e0228391.
6. Choy, K.T., Wong, A.Y., Kaewpreedee, P., Sia, S.F., Chen, D., Hui, K.P.Y., Chu, D.K.W., Chan, M.C.W., Cheung, P.P., Huang, X., Peiris, M., Yen, H.L., 2020. Remdesivir, lopinavir, emetine, and homoharringtonine inhibit SARS-CoV-2 replication in vitro. Antivir. Res. 178, 104786.
7. Cinatl, J., Morgenstern, B., Bauer, G., Chandra, P., Rabenau, H., Doerr, H.W., 2003. Treatment of SARS with human interferons. Lancet 362, 293–294.
8. Dahl, H., Linde, A., Strannegård, Ö., 2009. In vitro inhibition of SARS virus replication by human interferons. Scand. J. Infect. Dis. 36, 829–831.
9. de Wilde, A.H., Raj, V.S., Oudshoorn, D., Bestebroer, T.M., van Nieuwkoop, S., Limpens, R.W.A.L., Posthuma, C.C., van der Meer, Y., Bárcena, M., Haagmans, B.L., Snijder, E.J., van den Hoogen, B.G., 2013. MERS-coronavirus replication induces severe in vitro cytopathology and is strongly inhibited by cyclosporin A or interferonalpha treatment. J. Gen. Virol. 94, 1749–1760.
10. Felgenhauer, U., Schoen, A., Gad, H.H., Hartmann, R., Schaubmar, A.R., Failing, K., Drosten, C., Weber, F., 2020. Inhibition of SARS–CoV-2 by type I and type III interferons. J. Biol. Chem. 295, 13958–13964.
11. Gilead, 2021a. Gilead statement on Veklury^® (Remdesivir) and the SARS-CoV-2 omicron variant. Available: https://www.gilead.com/news-and-press/company-statements/gilead-statement-on-veklury-remdesivir-and-the-sars-cov-2-omicron-variant.(Accessed 1 December 2021).
12. Gilead, 2021b. Gilead's Veklury^® (Remdesivir) associated with a reduction in mortality rate in hospitalized patients with COVID-19 across three analyses of large retrospective real-world data sets. Available: https://www.gilead.com/news-andpress/press-room/press-releases/2021/6/gileads-veklury-remdesivir-associatedwith-a-reduction-in-mortality-rate-in-hospitalized-patients-with-covid19-a cross-three-analyses-of-large-ret. (Accessed 1 December 2021).
13. Gobeil, S.M., Janowska, K., McDowell, S., Mansouri, K., Parks, R., Manne, K., Stalls, V., Kopp, M.F., Henderson, R., Edwards, R.J., Haynes, B.F., Acharya, P., 2020. D614G mutation alters SARS-CoV-2 spike conformation and enhances protease cleavage at the S1/S2 junction. Cell Rep. 34, 108630.
14. Grein, J., Ohmagari, N., Shin, D., Diaz, G., Asperges, E., Castagna, A., Feldt, T., Green, G., Green, M.L., Lescure, F.X., Nicastri, E., Oda, R., Yo, K., QuirosRoldan, E., Studemeister, A., Redinski, J., Ahmed, S., Bernett, J., Chelliah, D., Chen, D., Chihara, S., Cohen, S.H., Cunningham, J., D’Arminio Monforte, A., Ismail, S., Kato, H., Lapadula, G., L’Her, E., Maeno, T., Majumder, S., Massari, M., Mora-Rillo, M., Mutoh, Y., Nguyen, D., Verweij, E., Zoufaly, A., Osinusi, A.O., DeZure, A., Zhao, Y., Zhong, L., Chokkalingam, A., Elboudwarej, E., Telep, L., Timbs, L., Henne, I., Sellers, S., Cao, H., Tan, S.K., Winterbourne, L., Desai, P., Mera, R., Gaggar, A., Myers, R.P., Brainard, D.M., Childs, R., Flanigan, T., 2020. Compassionate use of remdesivir for patients with severe covid-19. N. Engl. J. Med. 382, 2327–2336.
15. Hawkins, M.J., Borden, E.C., Merritt, J.A., Edwards, B.S., Ball, L.A., Grossbard, E., Simon, K.J., 1984. Comparison of the biologic effects of two recombinant human interferons alpha (rA and rD) in humans. J. Clin. Oncol. 2, 221–226.
16. Huang, Y.Q., Tang, S.Q., Xu, X.L., Zeng, Y.M., He, X.Q., Li, Y., Harypursat, V., Lu, Y.Q., Wan, Y., Zhang, L., Sun, Q.Z., Sun, N.N., Wang, G.X., Yang, Z.P., Chen, Y.K., 2020. No statistically apparent difference in antiviral effectiveness observed among ribavirin plus interferon-alpha, lopinavir/ritonavir plus interferon-alpha, and ribavirin plus lopinavir/ritonavir plus interferon-alpha in patients with mild to moderate coronavirus disease 2019: results of a randomized, open-labeled prospective study. Front. Pharmacol. 11, 1071.
17. Huang, X., Zhang, X., Wang, F., Wei, H., Ma, H., Sui, M., Lu, J., Wang, H., Dumler, J.S., Sheng, G., Xu, B., 2016. Clinical efficacy of therapy with recombinant human interferon alpha1b in hand, foot, and mouth disease with enterovirus 71 infection. PLoS One 11, e0148907.
18. Leung, K., Shum, M.H., Leung, G.M., Lam, T.T., Wu, J.T., 2021. Early transmissibility assessment of the N501Y mutant strains of SARS-CoV-2 in the United Kingdom, October to November 2020. Euro Surveill. 26, 2002106.
19. Li, M.F., Jin, Q., Hu, G., Guo, H.Y., Hou, Y.D., 1992. A novel variant of human interferon alpha 1 gene. Sci. China E B 35, 200–206.
20. Li, Q., Wu, J., Nie, J., Zhang, L., Hao, H., Liu, S., Zhao, C., Zhang, Q., Liu, H., Nie, L., Qin, H., Wang, M., Lu, Q., Li, X., Sun, Q., Liu, J., Zhang, L., Li, X., Huang, W., Wang, Y., 2020. The impact of mutations in SARS-CoV-2 spike on viral infectivity and antigenicity. Cell 182, 1284–1294 e9.
21. Li, Q.Q., Guan, X., Wu, P., Wang, X., Zhou, L., Tong, Y., Ren, R., Leung, K.S.M., Lau, E.H.Y., Wong, J.Y., Xing, X., Xiang, N., Wu, Y., Li, C., Chen, Q., Li, D., Liu, T., Zhao, J., Liu, M., Tu, W., Chen, C., Jin, L., Yang, R., Wang, Q., Zhou, S., Wang, R., Liu, H., Luo, Y., Liu, Y., Shao, G., Li, H., Tao, Z., Yang, Y., Deng, Z., Liu, B., Ma, Z., Zhang, Y., Shi, G., Lam, T.T.Y., Wu, J.T., Gao, G.F., Cowling, B.J., Yang, B., Leung, G.M., Feng, Z., 2020. Early transmission dynamics in Wuhan, China, of novel coronavirus-infected pneumonia. N. Engl. J. Med. 382, 1199–1207.
22. Loutfy, M.R., Blatt, L.M., Siminovitch, K.A., Ward, S., Wolff, B., Lho, H., Pham, D.H., Deif, H., LaMere, E.A., Chang, M., Kain, K.C., Farcas, G.A., Ferguson, P., Latchford, M., Levy, G., Dennis, J.W., Lai, E.K., Fish, E.N., 2003. Interferon alfacon-1 plus corticosteroids in severe acute respiratory syndrome: a preliminary study. JAMA 290, 3222–3228.
23. Pandit, A., Bhalani, N., Bhushan, B.L.S., Koradia, P., Gargiya, S., Bhomia, V., Kansagra, K., 2021. Efficacy and safety of pegylated interferon alfa-2b in moderate COVID-19: a phase II, randomized, controlled, open-label study. Int. J. Infect. Dis. 105, 516–521.
24. Reed, L.J., Muench, H., 1938. A simple method of estimating fifty per cent endpoints. Am. J. Epidemiol. 3, 493–497.
25. Sadler, A.J., Williams, B.R.G., 2008. Interferon-inducible antiviral effectors. Nat. Rev. Immunol. 8, 559–568.
26. Sainz, B., Mossel, E.C., Peters, C.J., Garry, R.F., 2004. Interferon-beta and interferongamma synergistically inhibit the replication of severe acute respiratory syndromeassociated coronavirus (SARS-CoV). Virology 329, 11–17.
27. Seidel, V., Feiterna-Sperling, C., Siedentopf, J.P., Hofmann, J., Henrich, W., Buhrer, C., Weizsacker, K., 2017. Intrauterine therapy of cytomegalovirus infection with valganciclovir: review of the literature. Med. Microbiol. Immunol. 206, 347–354.
28. Sheahan, T.P., Sims, A.C., Leist, S.R., Schäfer, A., Won, J., Brown, A.J., Montgomery, S.A., Hogg, A., Babusis, D., Clarke, M.O., Spahn, J.E., Bauer, L., Sellers, S., Porter, D., Feng, J.Y., Cihlar, T., Jordan, R., Denison, M.R., Baric, R.S., 2020. Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV. Nat. Commun. 11, 222.
29. Vanderheiden, A., Ralfs, P., Chirkova, T., Upadhyay, A.A., Zimmerman, M.G., Bedoya, S., Aoued, H., Tharp, G.M., Pellegrini, K.L., Manfredi, C., Sorscher, E., Mainou, B., Lobby, J.L., Kohlmeier, J.E., Lowen, A.C., Shi, P.Y., Menachery, V.D., Anderson, L.J., Grakoui, A., Bosinger, S.E., Suthar, M.S., 2020. Type I and type III interferons restrict SARS-CoV-2 infection of human airway epithelial cultures. J. Virol. 94 e00985-20.
30. Wang, M., Cao, R., Zhang, L., Yang, X., Liu, J., Xu, M., Shi, Z., Hu, Z., Zhong, W., Xiao, G., 2020. Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro. Cell Res. 30, 269–271.
31. Weisblum, Y., Schmidt, F., Zhang, F., DaSilva, J., Poston, D., Lorenzi, J.C., Muecksch, F., Rutkowska, M., Hoffmann, H.H., Michailidis, E., Gaebler, C., Agudelo, M., Cho, A., Wang, Z., Gazumyan, A., Cipolla, M., Luchsinger, L., Hillyer, C.D., Caskey, M., Robbiani, D.F., Rice, C.M., Nussenzweig, M.C., Hatziioannou, T., Bieniasz, P.D., 2020. Escape from neutralizing antibodies by SARS-CoV-2 spike protein variants. Elife 9, e61312.
32. Wu, J.T., Leung, K., Leung, G.M., 2020. Nowcasting and forecasting the potential domestic and international spread of the 2019-nCoV outbreak originating in Wuhan, China: a modelling study. Lancet 395, 689–697.
33. Zhang, L., Jackson, C.B., Mou, H., Ojha, A., Peng, H., Quinlan, B.D., Rangarajan, E.S., Pan, A., Vanderheiden, A., Suthar, M.S., Li, W., Izard, T., Rader, C., Farzan, M., Choe, H., 2020. SARS-CoV-2 spike-protein D614G mutation increases virion spike density and infectivity. Nat. Commun. 11, 6013.
34. Zumla, A., Chan, J.F.W., Azhar, E.I., Hui, D.S.C., Yuen, K.Y., 2016. Coronaviruses - drug discovery and therapeutic options. Nat. Rev. Drug Discov. 15, 327–347.
Proportional views
Electronic Supplementary Material

10.1016j.virs.2022.01.031-ESM.docx