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A single-domain antibody inhibits SFTSV and mitigates virus-induced pathogenesis in vivo
Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu
Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu
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Research Article Infectious disease Therapeutics

A single-domain antibody inhibits SFTSV and mitigates virus-induced pathogenesis in vivo

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Abstract

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a novel tick-borne bunyavirus that recently emerged in East Asian countries. SFTS is characterized by high fever, thrombocytopenia, leukopenia, multiorgan failure, and hemorrhage with case fatality rates of 6.3% to 30%. Neither antivirals nor vaccines are available at present. We previously demonstrated that neutralizing antibodies specific for SFTSV glycoprotein (Gn) played a vital role in the survival of patients with SFTS. Nanobodies from camels present unique properties, such as thermostability, high affinity, and low immunogenicity. In the current study, mammalian expressed SFTSV Gn was used to immunize a camel, and functional nanobodies were isolated from the B cell nanobody library constructed from the immunized animal. Clone SNB02 was selected for in-depth analysis for its inhibition of SFTSV replication both in vitro and in vivo. We showed that SNB02 potently inhibited SFTSV infection and prevented thrombocytopenia in a humanized mouse model and is a potential candidate for therapeutics.

Authors

Xilin Wu, Yanlei Li, Bilian Huang, Xiaohua Ma, Linjing Zhu, Nan Zheng, Shijie Xu, Waqas Nawaz, Changping Xu, Zhiwei Wu

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Figure 4

Characterization of SNB02.

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Characterization of SNB02.
(A) The purified SNB02 was determined by SDS-...
(A) The purified SNB02 was determined by SDS-PAGE under reducing condition (R) or nonreducing condition (NR). (B) Purity of SNB02 was determined by size exclusion high-performance liquid chromatography (SEC-HPLC) analysis. (C) sGn under reducing conditions (R) or nonreducing conditions (NR) was detected by Western blot with SNB02 and rabbit polyclonal antisera (Antisera) specific for sGn. (D) Kinetic binding curve of SNB02 with sGn. Different-colored curves represent different concentrations; sGn protein was bound by SNB02. (E) Neutralization activity of SNB02 against live SFTSV infection of PBMCs and Vero E6 cells, respectively. SNB-ctl, 1 VHH-HuFc antibody without binding with sGn, was taken as antibody negative control. The red and gray lines show the neutralized curves of SNB02 and SNB-ctl inhibiting SFTSV to infect PBMCs, respectively. The blue line and the black lines show the neutralized curves of SNB02 and SNB-ctl inhibiting SFTSV to infect Vero E6 cells. (F) Bioavailability and t1/2 of SNB02 in NCG-HuPBL mice. Data represent mean ± SEM. SNB02 group, n = 4 (2 M + 2 F); mock group, n = 2 (1 M + 1 F). All experiments of A, C, and E were repeated twice.

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