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| 1 | Rapid isolation and immune profiling of SARS-CoV-2 specific memory B cell in convalescent COVID-19 patients via LIBRA-seq显示文摘B cell response plays a critical role against SARS-CoV-2 infection.However,little is known about the diversity and frequency of the paired SARS-CoV-2 antigen-specific BCR repertoire after SARS-CoV-2 infection.Here,we performed single-cell RNA sequencing and VDJ sequencing using the memory and plasma B cells isolated from five convalescent COVID-19 patients,and analyzed the spectrum and transcriptional heterogeneity of antibody immune responses.Via linking BCR to antigen specificity through sequencing(LIBRA-seq),we identified a distinct activated memory B cell subgroup(CD11c^(high) CD95^(high))had a higher proportion of SARS-CoV-2 antigen-labeled cells compared with memory B cells.Our results revealed the diversity of paired BCR repertoire and the non-stochastic pairing of SARS-CoV-2 antigen-specific immunoglobulin heavy and light chains after SARS-CoV-2 infection.The public antibody clonotypes were shared by distinct convalescent individuals.Moreover,several antibodies isolated by LIBRA-seq showed high binding affinity against SARS-CoV-2 receptor-binding domain(RBD)or nucleoprotein(NP)via ELISA assay.Two RBD-reactive antibodies C14646P3S and C2767P3S isolated by LIBRA-seq exhibited high neutralizing activities against both pseudotyped and authentic SARS-CoV-2 viruses in vitro.Our study provides fundamental insights into B cell response following SARS-CoV-2 infection at the single-cell level. | Bing He Shuning Liu Yuanyuan Wang Mengxin Xu Wei Cai Jia Liu Wendi Bai Shupei Ye Yong Ma Hengrui Hu Huicui Meng Tao Sun Yanling Li Huanle Luo Mang Shi Xiangjun Du Wenjing Zhao Shoudeng Chen Jingyi Yang Haipeng Zhu Yusheng Jie Yuedong Yang Deyin Guo Qiao Wang Yuwen Liu Huimin Yan Manli Wang Yao-Qing Chen | 2021 | Signal Transduction and Targeted Therapy2021，6，6： | 2 |
| 2 | Hybridoma-derived neutralizing monoclonal antibodies against Beta and Delta variants of SARS-CoV-2 in vivo显示文摘Neutralizing monoclonal antibodies(mAb)are a major therapeutic strategy for the treatment of severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)infection.The continuous emergence of new SARS-CoV-2 variants worldwide has increased the urgency for the development of new mAbs.In this study,we immunized mice with the receptor-binding domain(RBD)of the SARS-CoV-2 prototypic strain(WIV04)and screened 35 RBDspecific mAbs using hybridoma technology.Results of the plaque reduction neutralization test showed that 25 of the mAbs neutralized authentic WIV04 strain infection.The 25 mAbs were divided into three categories based on the competitive enzyme-linked immunosorbent assay results.A representative mAb was selected from each category(RD4,RD10,and RD14)to determine the binding kinetics and median inhibitory concentration(IC_(50))of WIV04 and two variants of concern(VOC):B.1.351(Beta)and B.1.617.2(Delta).RD4 neutralized the B.1.617.2 variant with an IC50 of 2.67 ng/mL;however,it completely lost neutralizing activity against the B.1.351 variant.RD10 neutralized both variants with an IC50 exceeding 100 ng/mL;whereas RD14 neutralized two variants with a higher IC50(>1 mg/mL).Animal experiments were performed to evaluate the protective effects of RD4 and RD10 against various VOC infections.RD4 could protect Adv-hACE2 transduced mice from B.1.617.2 infection at an antibody concentration of 25 mg/kg,while RD10 could protect mice from B.1.351 infection at an antibody concentration of 75 mg/kg.These results highlight the potential for future modifications of the mAbs for practical use. | Qianran Wang Lu Peng Yanqiu Nie Yanni Shu Huajun Zhang Zidan Song Yufeng Li Hengrui Hu Liushuai Li Xi Wang Jia Liu Jiang Li Zhengli Shi Fei Deng Yu Guo Yiwu Zhou Bing Yan Zhihong Hu Manli Wang | 2023 | Virologica Sinica2023，38，2： | 0 |
| 3 | Construction and characterization of a synthesized herpes simplex virus H129-Syn-G2显示文摘Herpes simplex virus type 1(HSV-1)causes lifelong infections worldwide,and currently there is no efficient cure or vaccine.HSV-1-derived tools,such as neuronal circuit tracers and oncolytic viruses,have been used exten-sively;however,further genetic engineering of HSV-1 is hindered by its complex genome structure.In the present study,we designed and constructed a synthetic platform for HSV-1 based on H129-G4.The complete genome was constructed from 10 fragments through 3 rounds of synthesis using transformation-associated recombination(TAR)in yeast,and was named H129-Syn-G2.The H129-Syn-G2 genome contained two copies of the gfp gene and was transfected into cells to rescue the virus.According to growth curve assay and electron microscopy results,the synthetic viruses exhibited more optimized growth properties and similar morphogenesis compared to the parental virus.This synthetic platform will facilitate further manipulation of the HSV-1 genome for the devel-opment of neuronal circuit tracers,oncolytic viruses,and vaccines. | Han Xiao Hengrui Hu Yijia Guo Jiang Li Le Wen Wen-Bo Zeng Manli Wang Min-Hua Luo Zhihong Hu | 2023 | Virologica Sinica2023，38，3： | 0 |
| 4 | Construction and Characterization of a Novel Bacmid AcBac-Syn Based on a Synthesized Baculovirus Genome显示文摘Baculoviruses are large DNA viruses which have been widely used as expression vectors and biological insecticides.Homologous recombination and Bac-to-Bac system have been the main methods for manipulating the baculovirus genome.Recently, we generated a synthetic baculovirus Ac MNPV-WIV-Syn1 which fully resembled its parental virus Autographa californica multiple nucleopolyhedrovirus(Ac MNPV). Here, we report the modification of Ac MNPV-WIV-Syn1 into a novel bacmid, Ac Bac-Syn, which can be used as a backbone for Bac-to-Bac system. To achieve this, a vector contained a Lac Z:att Tn7 and egfp cassette was constructed, and recombined with a linearized Ac MNPV-WIV-Syn1 genome by transformation-associated recombination in yeast to generate bacmid Ac Bac-Syn. The bacmid was then transfected to insect cells and the rescued virus showed similar biological characteristics to the wild-type virus in terms of the kinetics of budded virus production, the morphology of occlusion bodies, and the oral infectivity in insect larvae. For demonstration, a red fluorescent protein gene Dsred was transposed into the att Tn7 site by conventional Bac-to-Bac method, and the transfection and infection assays showed that Ac Bac-Syn can be readily used for foreign gene insertion and expression.Ac Bac-Syn has several advantages over the conventional Ac MNPV bacmids, such as it contains an egfp reporter gene which facilitates visualization of virus propagation and titration;its DNA copy numbers could be induced to a higher level in E. coli;and the retaining of the native polyhedrin gene in the genome making it an attractive system for studying the functions of gene related to occlusion body assembly and oral infection. | Yu Shang Hengrui Hu Xi Wang Hualin Wang Fei Deng Manli Wang Zhihong Hu | 2021 | Virologica Sinica2021，36，6： | 0 |
| 5 | Comparative Antiviral Efficacy of Viral Protease Inhibitors against the Novel SARS-CoV-2 In Vitro显示文摘The recent outbreak of novel coronavirus pneumonia(COVID-19)caused by a new coronavirus has posed a great threat to public health.Identifying safe and effective antivirals is of urgent demand to cure the huge number of patients.Virusencoded proteases are considered potential drug targets.The human immunodeficiency virus protease inhibitors(lopinavir/ritonavir)has been recommended in the global Solidarity Trial in March launched by World Health Organization.However,there is currently no experimental evidence to support or against its clinical use.We evaluated the antiviral efficacy of lopinavir/ritonavir along with other two viral protease inhibitors in vitro,and discussed the possible inhibitory mechanism in silico.The in vitro to in vivo extrapolation was carried out to assess whether lopinavir/ritonavir could be effective in clinical.Among the four tested compounds,lopinavir showed the best inhibitory effect against the novel coronavirus infection.However,further in vitro to in vivo extrapolation of pharmacokinetics suggested that lopinavir/ritonavir could not reach effective concentration under standard dosing regimen[marketed as Kaletraò,contained lopinavir/ritonavir(200 mg/50 mg)tablets,recommended dosage is 400 mg/10 mg(2 tablets)twice daily].This research concluded that lopinavir/ritonavir should be stopped for clinical use due to the huge gap between in vitro IC50 and free plasma concentration.Nevertheless,the structure–activity relationship analysis of the four inhibitors provided further information for de novel design of future viral protease inhibitors of SARS-CoV-2. | Leike Zhang Jia Liu Ruiyuan Cao Mingyue Xu Yan Wu Weijuan Shang Xi Wang Huanyu Zhang Xiaming Jiang Yuan Sun Hengrui Hu Yufeng Li Gang Zou Min Zhang Lei Zhao Wei Li Xiaojia Guo Xiaomei Zhuang Xing-Lou Yang Zheng-Li Shi Fei Deng Zhihong Hu Gengfu Xiao Manli Wang Wu Zhong | 2020 | Virologica Sinica2020，35，6： | 0 |
| 6 | Systematic Analysis of 42 Autographa Californica Multiple Nucleopolyhedrovirus Genes Identifies An Additional Six Genes Involved in the Production of Infectious Budded Virus显示文摘Baculoviruses have been widely used as a vector for expressing foreign genes.Among numerous baculoviruses,Autographa californica multiple nucleopolyhedrovirus(AcMNPV)is the most frequently used and it encodes 155 open reading frames(ORFs).Here,we systematically investigated the impact of 42 genes of AcMNPV on the production of infectious budded viruses(BVs)by constructing gene-knockout bacmids and subsequently conducting transfection and infection assays.The results showed that among the 39 functionally unverified genes and 3 recently reported genes,36 are dispensable for infectious BV production,as the one-step growth curves of the gene-knockout viruses were not significantly different from those of the parental virus.Three genes(ac62,ac82 and ac106/107)are essential for infectious BV production,as deletions thereof resulted in complete loss of infectivity while the repaired viruses showed no significant difference in comparison to the parental virus.In addition,three genes(ac13,ac51 and ac120)are important but not essential for infectious BV production,as gene-knockout viruses produced significantly lower BV levels than that of the parental virus or repaired viruses.We then grouped the 155 AcMNPV genes into three categories(Dispensable,Essential,or Important for infectious BV production).Based on our results and previous publications,we constructed a schematic diagram of a potential mini-genome of AcMNPV,which contains only essential and important genes.The results shed light on our understanding of functional genomics of baculoviruses and provide fundamental information for future engineering of baculovirus expression system. | Tong Chen Xiaoyan Duan Hengrui Hu Yu Shang Yangbo Hu Fei Deng Hualin Wang Manli Wang Zhihong Hu | 2021 | Virologica Sinica2021，36，4： | 0 |
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