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51. Towards a solution to MERS: protective human monoclonal antibodies targeting different domains and functions of the MERS-coronavirus spike glycoprotein

Author

European Commission, Innovative Medicines Initiative, European Federation of Pharmaceutical Industries and Associations, Ministerio de Economía y Competitividad (España), National Institutes of Health (US), China Scholarship Council, Widjaja, Ivy, Wang, Chengbao, Haperen, Rien van, Gutierrez-Alvarez, Francisco J., Dieren, Brenda van, Okba, Nisreen M. A., Stalin Raj, V., Li, Wentao, Fernandez-Delgado, Raúl, Grosveld, Frank, Kuppeveld, Frank J. M. van, Haagmans, Bart L., Enjuanes Sánchez, Luis, Drabek, Dubravka, Bosch, Berend Jan, European Commission, Innovative Medicines Initiative, European Federation of Pharmaceutical Industries and Associations, Ministerio de Economía y Competitividad (España), National Institutes of Health (US), China Scholarship Council, Widjaja, Ivy, Wang, Chengbao, Haperen, Rien van, Gutierrez-Alvarez, Francisco J., Dieren, Brenda van, Okba, Nisreen M. A., Stalin Raj, V., Li, Wentao, Fernandez-Delgado, Raúl, Grosveld, Frank, Kuppeveld, Frank J. M. van, Haagmans, Bart L., Enjuanes Sánchez, Luis, Drabek, Dubravka, and Bosch, Berend Jan

Abstract

The Middle-East respiratory syndrome coronavirus (MERS-CoV) is a zoonotic virus that causes severe and often fatal respiratory disease in humans. Efforts to develop antibody-based therapies have focused on neutralizing antibodies that target the receptor binding domain of the viral spike protein thereby blocking receptor binding. Here, we developed a set of human monoclonal antibodies that target functionally distinct domains of the MERS-CoV spike protein. These antibodies belong to six distinct epitope groups and interfere with the three critical entry functions of the MERS-CoV spike protein: sialic acid binding, receptor binding and membrane fusion. Passive immunization with potently as well as with poorly neutralizing antibodies protected mice from lethal MERS-CoV challenge. Collectively, these antibodies offer new ways to gain humoral protection in humans against the emerging MERS-CoV by targeting different spike protein epitopes and functions.

Published
2019

57. Role of transcription regulatory sequence in regulation of gene expression and replication of porcine reproductive and respiratory syndrome virus

Author

China Postdoctoral Science Foundation, National Natural Science Foundation of China, European Commission, Ministerio de Ciencia e Innovación (España), Wang, Chengbao, Almazán, Fernando, Solá Gurpegui, Isabel, Enjuanes Sánchez, Luis, Abrahamyan, Levon, China Postdoctoral Science Foundation, National Natural Science Foundation of China, European Commission, Ministerio de Ciencia e Innovación (España), Wang, Chengbao, Almazán, Fernando, Solá Gurpegui, Isabel, Enjuanes Sánchez, Luis, and Abrahamyan, Levon

Abstract

In order to gain insight into the role of the transcription regulatory sequences (TRSs) in the regulation of gene expression and replication of porcine reproductive and respiratory syndrome virus (PRRSV), the enhanced green fluorescent protein (EGFP) gene, under the control of the different structural gene TRSs, was inserted between the N gene and 3′-UTR of the PRRSV genome and EGFP expression was analyzed for each TRS. TRSs of all the studied structural genes of PRRSV positively modulated EGFP expression at different levels. Among the TRSs analyzed, those of GP2, GP5, M, and N genes highly enhanced EGFP expression without altering replication of PRRSV. These data indicated that structural gene TRSs could be an extremely useful tool for foreign gene expression using PRRSV as a vector.

Published
2017

58. Characterization of the Interactome of the Porcine Reproductive and Respiratory Syndrome Virus Nonstructural Protein 2 Reveals the Hyper Variable Region as a Binding Platform for Association with 14–3–3 Proteins

Author

Xiao, Yihong, primary, Wu, Weining, additional, Gao, Jiming, additional, Smith, Nikki, additional, Burkard, Christine, additional, Xia, Dong, additional, Zhang, Minxia, additional, Wang, Chengbao, additional, Archibald, Alan, additional, Digard, Paul, additional, Zhou, En-min, additional, and Hiscox, Julian A., additional

Published
2016
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59. Rescue and evaluation of a recombinant PRRSV expressing porcine Interleukin-4

Author

Li, Zhijun, primary, Wang, Gang, additional, Wang, Yan, additional, Zhang, Chong, additional, Wang, Xinglong, additional, Huang, Baicheng, additional, Li, Qiongyi, additional, Li, Liangliang, additional, Xue, Biyun, additional, Ding, Peiyang, additional, Syed, Shahid Faraz, additional, Wang, Chengbao, additional, Cai, Xuehui, additional, and Zhou, En-Min, additional

Published
2015
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60. MicroRNA miR-24-3p Promotes Porcine Reproductive and Respiratory Syndrome Virus Replication through Suppression of Heme Oxygenase-1 Expression

Author

Xiao, Shuqi, primary, Wang, Xue, additional, Ni, Huaibao, additional, Li, Na, additional, Zhang, Angke, additional, Liu, Hongliang, additional, Pu, Fengxing, additional, Xu, Lele, additional, Gao, Jiming, additional, Zhao, Qin, additional, Mu, Yang, additional, Wang, Chengbao, additional, Sun, Yani, additional, Du, Taofeng, additional, Xu, Xingang, additional, Zhang, Gaiping, additional, Hiscox, Julian A., additional, Goodfellow, Ian G., additional, and Zhou, En-Min, additional

Published
2015
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61. Intracellular expression of an anti-idiotypic antibody single-chain variable fragment reduces porcine reproductive and respiratory syndrome virus infection in MARC-145 cells

Author

Li, Qiongyi, primary, Wang, Xiangpeng, additional, Wang, Chengbao, additional, Yu, Ying, additional, Wang, Gang, additional, Gao, Jiming, additional, Liu, Hongliang, additional, Xie, Huali, additional, Huang, Baicheng, additional, Li, Zhijun, additional, Kong, Ning, additional, Zhang, Gaiping, additional, Hsu, Walter H, additional, and Zhou, En-Min, additional

Published
2015
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62. GP5 expression in Marc-145 cells inhibits porcine reproductive and respiratory syndrome virus infection by inducing beta interferon activity

Author

Gao, Jiming, primary, Ji, Pengchao, additional, Zhang, Maodong, additional, Wang, Xiangpeng, additional, Li, Na, additional, Wang, Chengbao, additional, Xiao, Shuqi, additional, Mu, Yang, additional, Zhao, Qin, additional, Du, Taofeng, additional, Sun, Yani, additional, Hiscox, Julian A., additional, Zhang, Gaiping, additional, and Zhou, En-Min, additional

Published
2014
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63. Identification of an antigenic domain in the N-terminal region of avian hepatitis E virus (HEV) capsid protein that is not common to swine and human HEVs

Author

Wang, Lizhen, primary, Sun, Yani, additional, Du, Taofeng, additional, Wang, Chengbao, additional, Xiao, Shuqi, additional, Mu, Yang, additional, Zhang, Gaiping, additional, Liu, Lihong, additional, Widén, Frederik, additional, Hsu, Walter H., additional, Zhao, Qin, additional, and Zhou, En-Min, additional

Published
2014
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64. A novel porcine reproductive and respiratory syndrome virus vector system that stably expresses enhanced green fluorescent protein as a separate transcription unit

Author

Wang, Chengbao, Huang, Baicheng, Kong, Ning, Li, Qiongyi, Ma, Yuping, Li, Zhijun, Gao, Jiming, Zhang, Chong, Wang, Xiangpeng, Liang, Chao, Dang, Lu, Xiao, Shuqi, Mu, Yang, Zhao, Qin, Sun, Yani, Almazán, Fernando, Enjuanes Sánchez, Luis, Zhou, En-Min, Wang, Chengbao, Huang, Baicheng, Kong, Ning, Li, Qiongyi, Ma, Yuping, Li, Zhijun, Gao, Jiming, Zhang, Chong, Wang, Xiangpeng, Liang, Chao, Dang, Lu, Xiao, Shuqi, Mu, Yang, Zhao, Qin, Sun, Yani, Almazán, Fernando, Enjuanes Sánchez, Luis, and Zhou, En-Min

Abstract

Here we report the rescue of a recombinant porcine reproductive and respiratory syndrome virus (PRRSV) carrying an enhanced green fluorescent protein (EGFP) reporter gene as a separate transcription unit. A copy of the transcription regulatory sequence for ORF6 (TRS6) was inserted between the N protein and 3′-UTR to drive the transcription of the EGFP gene and yield a general purpose expression vector. Successful recovery of PRRSV was obtained using an RNA polymerase II promoter to drive transcription of the full-length virus genome, which was assembled in a bacterial artificial chromosome (BAC). The recombinant virus showed growth replication characteristics similar to those of the wild-type virus in the infected cells. In addition, the recombinant virus stably expressed EGFP for at least 10 passages. EGFP expression was detected at approximately 10 h post infection by live-cell imaging to follow the virus spread in real time and the infection of neighbouring cells occurred predominantly through cell-to-cell-contact. Finally, the recombinant virus generated was found to be an excellent tool for neutralising antibodies and antiviral compound screening. The newly established reverse genetics system for PRRSV could be a useful tool not only to monitor virus spread and screen for neutralising antibodies and antiviral compounds, but also for fundamental research on the biology of the virus.

Published
2013

65. Anti-idiotypic antibodies reduce efficacy of the attenuated vaccine against highly pathogenic PRRSV challenge

Author

Yu, Ying, primary, Cai, Xuehui, additional, Wang, Gang, additional, Kong, Ning, additional, Liu, Yonggang, additional, Xiao, Yihong, additional, Zhang, Chong, additional, Mu, Yang, additional, Xiao, Shuqi, additional, Zhao, Qin, additional, Wang, Chengbao, additional, Zhang, Gaiping, additional, Hiscox, Julian A, additional, and Zhou, En-Min, additional

Published
2014
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67. A novel porcine reproductive and respiratory syndrome virus vector system that stably expresses enhanced green fluorescent protein as a separate transcription unit

Author

Wang, Chengbao, primary, Huang, Baicheng, additional, Kong, Ning, additional, Li, Qiongyi, additional, Ma, Yuping, additional, Li, Zhijun, additional, Gao, Jiming, additional, Zhang, Chong, additional, Wang, Xiangpeng, additional, Liang, Chao, additional, Dang, Lu, additional, Xiao, Shuqi, additional, Mu, Yang, additional, Zhao, Qin, additional, Sun, Yani, additional, Almazan, Fernando, additional, Enjuanes, Luis, additional, and Zhou, En-Min, additional

Published
2013
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71. Microstructure and Mechanical Properties of FeCoNiCrAlxHigh-entropy Alloys by Selective Laser Melting

Author

Wen, Xuelong, Wang, Chengbao, Gong, Yadong, and Liu, Wenbo

Abstract

In this study, the thermal analysis theory of selective laser melting (SLM) was introduced, and different high-entropy alloy (HEA) specimens were prepared using the SLM technology. The effects of different powder sizes, elemental contents, and process parameters on the microstructure and mechanical properties of FeCoNiCrAlxHEA specimens fabricated using SLM were analyzed. Moreover, hardness and tensile tests of these high-entropy alloys were performed. The results showed that with increasing laser power and hatch spacing, the hardness of the specimens initially increased and subsequently decreased; it also increased with increasing scanning speed. The FeCoNiCrAl0.5HEA specimens prepared using fine powder exhibited better tensile properties, followed by FeCoNiCrAl0.8HEA. However, the FeCoNiCrAl0.5HEA prepared using coarse powder exhibited the poorest tensile properties. A comparison of the tensile properties of the specimens at different heights revealed that the specimens formed at the middle height exhibited improved tensile properties.

Published
2023
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72. Intracellular Expression of an Anti-Idiotypic Antibody Single-Chain Variable Fragment Reduces Porcine Reproductive and Respiratory Syndrome Virus Infection in MARC-145 Cells

Author

Li, Qiongyi, Wang, Xiangpeng, Wang, Chengbao, Yu, Ying, Wang, Gang, Gao, Jiming, Liu, Hongliang, Xie, Huali, Huang, Baicheng, Li, Zhijun, Kong, Ning, Zhang, Gaiping, Hsu, Walter H, and Zhou, En-Min

Abstract

Background Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome; it is one of the most economically important viral diseases affecting the swine industry worldwide. At present, neither live-attenuated nor inactivated PRRSV vaccines can provide sustainable disease control. Our previous studies have demonstrated that PRRSV infection can produce the auto-anti-idiotypic antibodies (aAb2s) specific to the idiotypic antibodies against PRRSV GP5, which plays an important role in the host immune responses to PRRSV infection. In the present study, a single-chain variable antibody fragment (scFv) from the monoclonal anti-idiotypic antibody specific for the idiotypic antibody against GP5 was expressed in MARC-145 cells and its effect on virus infection in vitrowas evaluated.Methods An scFv was constructed from the anti-idiotypic antibody (Mab2-5G2) and was named 5G2scFv. The lentiviral vector system was used as a vehicle to deliver 5G2scFv into MARC-145 cells. The effect of 5G2scFv expression in MARC-145 was analysed by determining the PRRSV N protein level and the virus titre in the supernatant. Virus attachment and the level of type I interferon (IFN) were determined to elucidate the mechanism of the scFv effect.Results 5G2scFv was delivered in MARC-145 cells using the lentiviral vector system as confirmed by the western blot and indirect immunofluorescence assays. The PRRSV challenge experiments demonstrated that expressed 5G2scFv in MARC-145 strongly reduced PRRSV infection and replication by inhibiting protein synthesis and progeny virus production. This effect was not due to the change of viability or virus binding, but increased IFN-a at messenger RNA and protein levels.Conclusions The expression of the anti-idiotypic antibody 5G2scFv in MARC-145 cells has the interferential effect on PRRSV infection in the cells by induction of IFN-a, which provides a novel therapeutic approach for PRRSV infection.

Published
2016
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73. Transparent and flexible electromagnetic interference shielding film based on Ag nanowires/ionic liquids/thermoplastic polyurethane ternary composites

Author

Wang, Chengbao, Guo, Yabin, Chen, Jianwen, and Zhu, Yutian

Abstract

Flexible and transparent electromagnetic interference (EMI) shielding films are desired in various fields. Herein, we propose a simple but effective approach to fabricating the flexible, transparent, and high EMI shielding films based on Ag nanowires (AgNWs)/ionic liquids (ILs)/thermoplastic polyurethane (TPU) ternary composites. Attributed to the formation of the low-density but highly interconnected AgNW network as well as the synergistic effect of AgNWs and ILs on composite conductivity, the as-prepared AgNWs/ILs/TPU film has a transmittance of 67% and an EMI shielding effectiveness of ∼26.8 dB at 10 GHz. Because of the low areal density of AgNWs, the absolute shielding effectiveness (SSE/t) of AgNWs/ILs/TPU film even reaches to 1.32 × 106 dB cm2 g−1at 10 GHz, which is superior to most reported works. More interestingly, the as-prepared film also exhibits good durability and stability in the EMI shielding properties even after 1000 uninterrupted bending cycles or long-term soaking in acidic or alkali solution. These outstanding performances endow the as-prepared film with great potential in flexible and transparent EMI shielding scenarios.

Published
2022
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74. Efficacy and Safety of Yanggan Jian in Hepatitis B Virus-related Decompensated Cirrhosis: A Randomized, Double-blind, Controlled Trial.

Author

Chen L, Zhao C, Yao W, Liu W, Zhang H, Mu Y, Cai H, Xue D, Wang C, Wang W, Lin Y, Chen J, and Liu P

Abstract

Background and Aims: The aim was to evaluate the efficacy and safety of Yanggan Jian (YGJ) in HBV-infected patients with decompensated cirrhosis., Methods: This randomized, double-blind controlled trial enrolled 160 patients with HBV-related decompensated cirrhosis who were already receiving or about to start antiviral therapy. Patients were randomly assigned to receive YGJ or placebo for 24 weeks, and were followed-up to 36 weeks. The primary outcome was the proportion of patients with a ≥2 point reduction in Child-Turcotte-Pugh (CTP) score from baseline at week 24. Secondary outcomes were CTP class and score, serum liver function indices, mortality, incidence of hepatocellular carcinoma and variceal bleeding., Results: The proportion of patients with a CTP score reduction ≥2 was significantly greater in the YGJ than in the placebo group ( p =0.009); the percentage of patients with CTP class C was significantly less than that in the placebo group ( p <0.05), and the YGJ group had a significantly greater mean change from baseline in CTP score at week 24 ( p =0.034). The improvement in measured values and change from baseline of prothrombin time, serum albumin, platelets, cholinesterase, international normalized ratio, and activated partial thromboplastin time were significantly better with YGJ than with placebo. Between-group differences in cumulative rates of variceal bleeding, hepatocellular carcinoma, death, or the frequency of any adverse event (AE), AEs related to treatment, or discontinuation because of AEs were not significant., Conclusions: YGJ significantly improved CTP scores and hepatic synthetic and reserve function in patients with HBV-related decompensated cirrhosis, and was safe and well tolerated., Competing Interests: The authors have no conflict of interests related to this publication., (© 2023 Authors.)

Published
2023
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75. [Development of porcine induced pluripotent stem cells with a CD163 reporter system].

Author

Yue W, Zhang J, Wu X, Yang X, Shen Q, Yu S, Zhu Z, Wang C, Zhang S, and Hua J

Subjects
Swine, Animals, Receptors, Cell Surface genetics, Antigens, CD genetics, Antigens, CD metabolism, CD163 Antigen, Induced Pluripotent Stem Cells metabolism, Porcine respiratory and reproductive syndrome virus genetics
Abstract

As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163 . Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.

Published
2023
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