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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 1

Characterization of antisera specific for sGn.

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Characterization of antisera specific for sGn.
(A) The experimental sche...
(A) The experimental schedule of immunization; (B) sGn protein with his tag (sGn-his) or rabbit Fc tag (sGn-rFc) were detected by SDS-PAGE. (C) The titer of antisera was evaluated after the fourth immunization in camel receiving sGn. The y axis represents the absorbance at 450 nm; x axis, the antisera dilution fold. Antisera binding with sGn and coating buffer were labeled as sGn with green line and labeled as blank with black line, respectively. # represents the antiserum titer. (D) Determination of the specificity of antisera by immunofluorescence assay. 293TT cells were transiently transfected with Gn plasmid of SFTSV subtypes A, B, C, D, and E, and the cells were stained with the camel fourth antisera and sera preimmunization (Pre-bleed) for Gn (green). Mock served as a cell control without plasmid transfection. Images were visualized under the ×20 objective. All experiments of B–D were repeated twice.

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