Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization
Abstract Background Porcine epidemic diarrhea virus (PEDV) is emerging as a pathogenic coronavirus that causes a huge economic burden to the swine industry. Interaction of the viral spike (S) surface glycoprotein with the host cell receptor is recognized as the first step of infection and is the mai...
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| Format: | Article |
| Language: | English |
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BMC
2018-08-01
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| Series: | Virology Journal |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12985-018-1042-3 |
| _version_ | 1811240509881974784 |
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| author | Lang Gong Ying Lin Jianru Qin Qianniu Li Chunyi Xue Yongchang Cao |
| author_facet | Lang Gong Ying Lin Jianru Qin Qianniu Li Chunyi Xue Yongchang Cao |
| author_sort | Lang Gong |
| collection | DOAJ |
| description | Abstract Background Porcine epidemic diarrhea virus (PEDV) is emerging as a pathogenic coronavirus that causes a huge economic burden to the swine industry. Interaction of the viral spike (S) surface glycoprotein with the host cell receptor is recognized as the first step of infection and is the main determinant of virus tropism. The mechanisms by which neutralizing antibodies inhibit PEDV have not been defined. Isolating PEDV neutralizing antibodies are crucial to identifying the receptor-binding domains of the viral spike and elucidating the mechanism of protection against PEDV infection. Methods B cell hybridoma technique was used to generate hybridoma cells that secrete specific antibodies. E.coli prokaryotic expression system and Bac-to-Bac expression system were used to identify the target protein of each monoclonal antibody. qPCR was performed to analyze PEDV binding to Vero E6 cells with neutralizing antibody. Results We identified 10 monoclonal antibodies using hybridoma technology. Remarkably, 4 mAbs (designed 2G8, 2B11, 3D9, 1E3) neutralized virus infection potently, of which 2B11 and 1E3 targeted the conformational epitope of the PEDV S protein. qPCR results showed that both 2B11 and 2G8 blocked virus entry into Vero cells. Conclusion The data suggested that PEDV neutralizing antibody inhibited virus infection by binding to infectious virions, which could work as a tool to find the receptor-binding domains. |
| first_indexed | 2024-04-12T13:21:37Z |
| format | Article |
| id | doaj.art-5506f06db6524114ba2f9362ccdd8a69 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-12T13:21:37Z |
| publishDate | 2018-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-5506f06db6524114ba2f9362ccdd8a692022-12-22T03:31:27ZengBMCVirology Journal1743-422X2018-08-011511910.1186/s12985-018-1042-3Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalizationLang Gong0Ying Lin1Jianru Qin2Qianniu Li3Chunyi Xue4Yongchang Cao5State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityState Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityState Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityState Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityState Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityState Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen UniversityAbstract Background Porcine epidemic diarrhea virus (PEDV) is emerging as a pathogenic coronavirus that causes a huge economic burden to the swine industry. Interaction of the viral spike (S) surface glycoprotein with the host cell receptor is recognized as the first step of infection and is the main determinant of virus tropism. The mechanisms by which neutralizing antibodies inhibit PEDV have not been defined. Isolating PEDV neutralizing antibodies are crucial to identifying the receptor-binding domains of the viral spike and elucidating the mechanism of protection against PEDV infection. Methods B cell hybridoma technique was used to generate hybridoma cells that secrete specific antibodies. E.coli prokaryotic expression system and Bac-to-Bac expression system were used to identify the target protein of each monoclonal antibody. qPCR was performed to analyze PEDV binding to Vero E6 cells with neutralizing antibody. Results We identified 10 monoclonal antibodies using hybridoma technology. Remarkably, 4 mAbs (designed 2G8, 2B11, 3D9, 1E3) neutralized virus infection potently, of which 2B11 and 1E3 targeted the conformational epitope of the PEDV S protein. qPCR results showed that both 2B11 and 2G8 blocked virus entry into Vero cells. Conclusion The data suggested that PEDV neutralizing antibody inhibited virus infection by binding to infectious virions, which could work as a tool to find the receptor-binding domains.http://link.springer.com/article/10.1186/s12985-018-1042-3Porcine epidemic diarrhea virusS proteinNeutralizing antibodyAttachment |
| spellingShingle | Lang Gong Ying Lin Jianru Qin Qianniu Li Chunyi Xue Yongchang Cao Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization Virology Journal Porcine epidemic diarrhea virus S protein Neutralizing antibody Attachment |
| title | Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| title_full | Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| title_fullStr | Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| title_full_unstemmed | Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| title_short | Neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| title_sort | neutralizing antibodies against porcine epidemic diarrhea virus block virus attachment and internalization |
| topic | Porcine epidemic diarrhea virus S protein Neutralizing antibody Attachment |
| url | http://link.springer.com/article/10.1186/s12985-018-1042-3 |
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