Mutation of FMDV L pro H138 residue drives viral attenuation in cell culture and in vivo in swine.

Autor: Azzinaro PA; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., Medina GN; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States.; National Bio and Agro-Defense Facility (NBAF), Agricultural Research Service (ARS), U.S. Department of Agriculture (USDA), Manhattan, KS, United States., Rai D; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States.; Pfizer Worldwide Research, Development and Medical, Pearl River, NY, United States., Ramirez-Medina E; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., Spinard E; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., Rodriguez-Calzada M; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States.; Oak Ridge Institute for Science and Education, PIADC Research Participation Program, Oak Ridge, TN, United States., Zhu J; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., Rieder E; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., de Los Santos T; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States., Díaz-San Segundo F; Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, NY, United States.
Jazyk: angličtina
Zdroj: Frontiers in veterinary science [Front Vet Sci] 2022 Oct 31; Vol. 9, pp. 1028077. Date of Electronic Publication: 2022 Oct 31 (Print Publication: 2022).
DOI: 10.3389/fvets.2022.1028077
Abstrakt: The foot-and-mouth disease virus (FMDV) leader proteinase (L pro ) 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 L pro 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 L pro 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-L H138L 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-L H138L 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 L pro 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.
Competing Interests: Author DR is currently employed by the company Pfizer Worldwide Research. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
(Copyright © 2022 Azzinaro, Medina, Rai, Ramirez-Medina, Spinard, Rodriguez-Calzada, Zhu, Rieder, de los Santos and Díaz-San Segundo.)
Databáze: MEDLINE
Externí odkaz: