dab7aecab249c7c9d08e27a40501bf04 pmc_10819402.pdf 849e3c935696bbe13e35e10433387fe587ecae07 pmc_10819402.pdf b2eade73a250e526a5ada6b1165c9d15a8ff747be6d6da5c730ed0101ee0e225 pmc_10819402.pdf Title: Safety and Efficacy upon Infection in Sheep with Rift Valley Fever Virus ZH548-rA2, a Triple Mutant Rescued Virus Subject: The introduction of three single nucleotide mutations into the genome of the virulent RVFV ZH548 strain allows for the rescue of a fully attenuated virus in mice (ZH548-rA2). These mutations are located in the viral genes encoding the RdRp and the non-structural protein NSs. This paper shows the results obtained after the subcutaneous inoculation of ZH548-rA2 in adult sheep and the subsequent challenge with the parental virus (ZH548-rC1). Inoculation with the ZH548-rA2 virus caused no detectable clinical or pathological effect in sheep, whereas inoculation of the parental rC1 virus caused lesions compatible with viral infection characterised by the presence of scattered hepatic necrosis. Viral infection was confirmed via immunohistochemistry, with hepatocytes within the necrotic foci appearing as the main cells immunolabelled against viral antigen. Furthermore, the inoculation of sheep with the rA2 virus prevented the liver damage expected after rC1 virus inoculation, suggesting a protective efficacy in sheep which correlated with the induction of both humoral and cell-mediated immune responses. Keywords: Rift Valley fever virus; attenuating mutations; vaccine efficacy; sheep pathology Author: Sandra Moreno, Gema Lorenzo, Álvaro López-Valiñas, Nuria de la Losa, Celia Alonso, Elena Charro, José I. Núñez, Pedro J. Sánchez-Cordón, Belén Borrego and Alejandro Brun Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri Jan 5 07:04:13 2024 CET ModDate: Fri Jan 5 07:07:02 2024 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 4929993 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- RUGTHW+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 XOBIIF+VnURWPalladioL Type 1 Custom yes yes yes 16 0 RNGDIR+URWPalladioL-Bold Type 1 Custom yes yes yes 21 0 SLXOTU+URWPalladioL-Roma Type 1 Custom yes yes yes 27 0 REMKTY+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 HOWKDX+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 48 0 OQIVQT+EURM10 Type 1 Builtin yes yes yes 65 0 VIWIOF+CMSY10 Type 1 Builtin yes yes yes 73 0 GEWEQC+PazoMath-Italic Type 1 Builtin yes yes yes 84 0 WABLSD+SFRM1000 Type 1 Custom yes yes yes 89 0 CIDFont+F3 CID TrueType Identity-H yes no yes 100 0 CIDFont+F1 CID TrueType Identity-H yes no yes 108 0 CIDFont+F2 CID TrueType Identity-H yes no yes 116 0 CIDFont+F4 CID TrueType Identity-H yes no yes 124 0 CIDFont+F5 CID TrueType Identity-H yes no yes 132 0 CIDFont+F3 CID TrueType Identity-H yes no yes 143 0 CIDFont+F1 CID TrueType Identity-H yes no yes 151 0 CIDFont+F2 CID TrueType Identity-H yes no yes 159 0 CIDFont+F6 CID TrueType Identity-H yes no yes 167 0 CIDFont+F3 CID TrueType Identity-H yes no yes 263 0 CIDFont+F1 CID TrueType Identity-H yes no yes 271 0 CIDFont+F2 CID TrueType Identity-H yes no yes 279 0 CIDFont+F7 CID TrueType Identity-H yes no yes 287 0 CIDFont+F8 CID TrueType Identity-H yes no yes 295 0 CIDFont+F3 CID TrueType Identity-H yes no yes 371 0 CIDFont+F1 CID TrueType Identity-H yes no yes 379 0 CIDFont+F2 CID TrueType Identity-H yes no yes 387 0 CIDFont+F3 CID TrueType Identity-H yes no yes 399 0 CIDFont+F1 CID TrueType Identity-H yes no yes 407 0 CIDFont+F2 CID TrueType Identity-H yes no yes 415 0 CIDFont+F7 CID TrueType Identity-H yes no yes 423 0 CIDFont+F8 CID TrueType Identity-H yes no yes 431 0 CIDFont+F3 CID TrueType Identity-H yes no yes 507 0 CIDFont+F1 CID TrueType Identity-H yes no yes 515 0 CIDFont+F2 CID TrueType Identity-H yes no yes 523 0 CIDFont+F3 CID TrueType Identity-H yes no yes 537 0 CIDFont+F1 CID TrueType Identity-H yes no yes 545 0 CIDFont+F2 CID TrueType Identity-H yes no yes 553 0 CIDFont+F7 CID TrueType Identity-H yes no yes 561 0 CIDFont+F6 CID TrueType Identity-H yes no yes 569 0 CIDFont+F3 CID TrueType Identity-H yes no yes 646 0 CIDFont+F1 CID TrueType Identity-H yes no yes 654 0 CIDFont+F2 CID TrueType Identity-H yes no yes 662 0 CIDFont+F7 CID TrueType Identity-H yes no yes 670 0 CIDFont+F6 CID TrueType Identity-H yes no yes 678 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-03-09 02:33:26 CET RepresentationInformation: pmc_10819402.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-03-01 02:38:13 CET Size: 4929993 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 879 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Cells and Viruses Destination: subsection.2.1 Item: Title: Animal Study Design Destination: subsection.2.2 Item: Title: Blood Chemistry Destination: subsection.2.3 Item: Title: Virus Detection in Blood Samples Destination: subsection.2.4 Item: Title: Sequencing Analysis Destination: subsection.2.5 Item: Title: Tissue Sampling, Histopathological and Immunohistochemical Analysis Destination: subsection.2.6 Item: Title: Expression of Recombinant Antigens Destination: subsection.2.7 Item: Title: Antibody Assays Destination: subsection.2.8 Item: Title: Interferon- Detection in Sheep Plasma and Upon In Vitro Re-Stimulation Analysis Destination: subsection.2.9 Item: Title: Results Destination: section.3 Children: Item: Title: Clinical and Pathological Findings Destination: subsection.3.1 Item: Title: Analysis of Viremia Levels Via RT-qPCR and Virus Isolation Destination: subsection.3.2 Item: Title: Analysis of Immune Responses Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: appendix.A. Info: Title: Safety and Efficacy upon Infection in Sheep with Rift Valley Fever Virus ZH548-rA2, a Triple Mutant Rescued Virus Author: Sandra Moreno, Gema Lorenzo, Álvaro López-Valiñas, Nuria de la Losa, Celia Alonso, Elena Charro, José I. Núñez, Pedro J. Sánchez-Cordón, Belén Borrego and Alejandro Brun Subject: The introduction of three single nucleotide mutations into the genome of the virulent RVFV ZH548 strain allows for the rescue of a fully attenuated virus in mice (ZH548-rA2). These mutations are located in the viral genes encoding the RdRp and the non-structural protein NSs. This paper shows the results obtained after the subcutaneous inoculation of ZH548-rA2 in adult sheep and the subsequent challenge with the parental virus (ZH548-rC1). Inoculation with the ZH548-rA2 virus caused no detectable clinical or pathological effect in sheep, whereas inoculation of the parental rC1 virus caused lesions compatible with viral infection characterised by the presence of scattered hepatic necrosis. Viral infection was confirmed via immunohistochemistry, with hepatocytes within the necrotic foci appearing as the main cells immunolabelled against viral antigen. Furthermore, the inoculation of sheep with the rA2 virus prevented the liver damage expected after rC1 virus inoculation, suggesting a protective efficacy in sheep which correlated with the induction of both humoral and cell-mediated immune responses. 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