Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine
The foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) is a papain like protease that cleaves the viral polyprotein and several host factors affecting host cell translation and induction of innate immunity. Introduction of Lpro mutations ablating catalytic activity is not tolerated by the...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2022.1028077/full |
| _version_ | 1797991735205298176 |
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| author | Paul A. Azzinaro Gisselle N. Medina Gisselle N. Medina Devendra Rai Devendra Rai Elizabeth Ramirez-Medina Edward Spinard Monica Rodriguez-Calzada Monica Rodriguez-Calzada James Zhu Elizabeth Rieder Teresa de los Santos Fayna Díaz-San Segundo |
| author_facet | Paul A. Azzinaro Gisselle N. Medina Gisselle N. Medina Devendra Rai Devendra Rai Elizabeth Ramirez-Medina Edward Spinard Monica Rodriguez-Calzada Monica Rodriguez-Calzada James Zhu Elizabeth Rieder Teresa de los Santos Fayna Díaz-San Segundo |
| author_sort | Paul A. Azzinaro |
| collection | DOAJ |
| description | The foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) is a papain like protease that cleaves the viral polyprotein and several host factors affecting host cell translation and induction of innate immunity. Introduction of Lpro mutations ablating catalytic activity is not tolerated by the virus, however, complete coding sequence deletion or introduction of targeted amino acid substitutions can render viable progeny. In proof-of-concept studies, we have previously identified and characterized FMDV Lpro mutants that are attenuated in cell culture and in animals, while retaining their capacity for inducing a strong adaptive immunity. By using molecular modeling, we have now identified a His residue (H138), that resides outside the substrate binding and catalytic domain, and is highly conserved across serotypes. Mutation of H138 renders possible FMDV variants of reduced virulence in vitro and in vivo. Kinetics studies showed that FMDV A12-LH138L mutant replicates similarly to FMDV A12-wild type (WT) virus in cells that do not offer immune selective pressure, but attenuation is observed upon infection of primary or low passage porcine epithelial cells. Western blot analysis on protein extracts from these cells, revealed that while processing of translation initiation factor eIF-4G was slightly delayed, no degradation of innate sensors or effector molecules such as NF-κB or G3BP2 was observed, and higher levels of interferon (IFN) and IFN-stimulated genes (ISGs) were induced after infection with A12-LH138L as compared to WT FMDV. Consistent with the results in porcine cells, inoculation of swine with this mutant resulted in a mild, or in some cases, no clinical disease but induction of a strong serological adaptive immune response. These results further support previous evidence that Lpro is a reliable target to derive numerous viable FMDV strains that alone or in combination could be exploited for the development of novel FMD vaccine platforms. |
| first_indexed | 2024-04-11T08:56:59Z |
| format | Article |
| id | doaj.art-2e733e01cba842d290abcaa40b87ced2 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-04-11T08:56:59Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Veterinary Science |
| spelling | doaj.art-2e733e01cba842d290abcaa40b87ced22022-12-22T04:33:13ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692022-10-01910.3389/fvets.2022.10280771028077Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swinePaul A. Azzinaro0Gisselle N. Medina1Gisselle N. Medina2Devendra Rai3Devendra Rai4Elizabeth Ramirez-Medina5Edward Spinard6Monica Rodriguez-Calzada7Monica Rodriguez-Calzada8James Zhu9Elizabeth Rieder10Teresa de los Santos11Fayna Díaz-San Segundo12Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesNational Bio and Agro-Defense Facility (NBAF), Agricultural Research Service (ARS), U.S. Department of Agriculture (USDA), Manhattan, KS, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPfizer Worldwide Research, Development and Medical, Pearl River, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesOak Ridge Institute for Science and Education, PIADC Research Participation Program, Oak Ridge, TN, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesPlum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United StatesThe foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) is a papain like protease that cleaves the viral polyprotein and several host factors affecting host cell translation and induction of innate immunity. Introduction of Lpro mutations ablating catalytic activity is not tolerated by the virus, however, complete coding sequence deletion or introduction of targeted amino acid substitutions can render viable progeny. In proof-of-concept studies, we have previously identified and characterized FMDV Lpro mutants that are attenuated in cell culture and in animals, while retaining their capacity for inducing a strong adaptive immunity. By using molecular modeling, we have now identified a His residue (H138), that resides outside the substrate binding and catalytic domain, and is highly conserved across serotypes. Mutation of H138 renders possible FMDV variants of reduced virulence in vitro and in vivo. Kinetics studies showed that FMDV A12-LH138L mutant replicates similarly to FMDV A12-wild type (WT) virus in cells that do not offer immune selective pressure, but attenuation is observed upon infection of primary or low passage porcine epithelial cells. Western blot analysis on protein extracts from these cells, revealed that while processing of translation initiation factor eIF-4G was slightly delayed, no degradation of innate sensors or effector molecules such as NF-κB or G3BP2 was observed, and higher levels of interferon (IFN) and IFN-stimulated genes (ISGs) were induced after infection with A12-LH138L as compared to WT FMDV. Consistent with the results in porcine cells, inoculation of swine with this mutant resulted in a mild, or in some cases, no clinical disease but induction of a strong serological adaptive immune response. These results further support previous evidence that Lpro is a reliable target to derive numerous viable FMDV strains that alone or in combination could be exploited for the development of novel FMD vaccine platforms.https://www.frontiersin.org/articles/10.3389/fvets.2022.1028077/fullFMDVfoot-and-mouth disease virusleader proteinaseLproattenuated virus |
| spellingShingle | Paul A. Azzinaro Gisselle N. Medina Gisselle N. Medina Devendra Rai Devendra Rai Elizabeth Ramirez-Medina Edward Spinard Monica Rodriguez-Calzada Monica Rodriguez-Calzada James Zhu Elizabeth Rieder Teresa de los Santos Fayna Díaz-San Segundo Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine Frontiers in Veterinary Science FMDV foot-and-mouth disease virus leader proteinase Lpro attenuated virus |
| title | Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine |
| title_full | Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine |
| title_fullStr | Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine |
| title_full_unstemmed | Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine |
| title_short | Mutation of FMDV Lpro H138 residue drives viral attenuation in cell culture and in vivo in swine |
| title_sort | mutation of fmdv lpro h138 residue drives viral attenuation in cell culture and in vivo in swine |
| topic | FMDV foot-and-mouth disease virus leader proteinase Lpro attenuated virus |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2022.1028077/full |
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