Alterations in the equine herpesvirus type-1 (EHV-1) strain RacH during attenuation.
Abstract: The equine herpesvirus type-1 modified live-vaccine strain RacH (256th passage on porcine embryonic kidney cells) was investigated by restriction-enzyme analysis and compared to representative plaque isolates of the 12th passage (RacL11, RacL22) and 185th passage (RacM24, RacM36). The restriction patterns of all Rac plaque isolates differed compared with reference strain Ab4. The left UL terminus was shortened by 0.1 kbp and a missing BamHI site led to the fusion of the f and t fragments. In some Rac derivatives, losses of restriction sites without deletions were observed: 1. One BamHI site located in the ribosyl reductase gene was missing in RacH, RacM24, RacM36, and RacL22; and 2. An SalI site mapping to the gp14 (gB) gene was absent in RacM24, RacM36 and RacH. An identical deletion of 0.85 kbp in size was found in both copies of the inverted repeat (IR) regions of RacH. The deletion was present only in the terminal IR of the medium-passage derivative RacM36. By contrast, in the genomes of the apathogenic RacM24, as well as the pathogenic plaque isolates RacL11 and RacL22, no deletions in the IRs were detectable. Nucleotide-sequence and Northern-blot analyses revealed that the deletions led to the elimination of one or both copies of the gene 67 (IR6) open-reading frame in RacM36 and RacH and affected the gene 68 (EUS1) in RacH.
Publication Date: 1996-03-01 PubMed ID: 8919964DOI: 10.1111/j.1439-0450.1996.tb00282.xGoogle Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the changes in equine herpesvirus type-1 strain RacH during attenuation and compares it with other strains, discovering notable variations in its genetic structure, and specifically in its terminal inverted repeat regions.
Methodology of the Study
- The study involved the examination of the equine herpesvirus type-1 modified live-vaccine strain RacH. This particular strain was on its 256th passage on porcine embryonic kidney cells.
- In aid of restriction-enzyme analysis, RacH was compared to representative plaque isolates of the 12th passage (RacL11, RacL22) and 185th passage (RacM24, RacM36).
Findings of the Study
- The researchers found that the restriction patterns of all Rac plaque isolates differed when compared with reference strain Ab4.
- The left UL terminus of the strain was reduced by 0.1 kbp, and a missing BamHI site led to the fusion of the f and t fragments.
- The study discovered that several Rac derivatives exhibited loss of restriction sites without any deletions.
- One such instance was the missing BamHI site located in the ribosyl reductase gene in several strains like RacH, RacM24, RacM36, and RacL22.
- In another instance, an SalI site, mapping to the gp14 (gB) gene, was not present in strains RacM24, RacM36, and RacH.
- Upon analysis, the research team found an identical deletion of 0.85 kbp in both copies of the inverted repeat (IR) regions of RacH. This particular deletion was present only in the terminal IR of the medium-passage derivative RacM36.
- The genomes of strains RacM24, RacL11, and RacL22, on the other hand, showed no deletions in the IRs.
- Description of the mutations were confirmed through nucleotide-sequence and Northern-blot analyses, revealing that the deletions led to the disappearance of one or both copies of the gene 67 (IR6) open-reading frame in RacM36 and RacH. Additionally, these deletions affected the gene 68 (EUS1) in RacH.
Implications of the Study
- The results give a closer look into the genetic changes that occur in the equine herpesvirus type-1 strain RacH during attenuation.
- The changes found could potentially affect the virus’s capacity to cause disease, which is crucial for understanding its pathogenic behavior and for the development of future therapies.
Cite This Article
APA
Hübert PH, Birkenmaier S, Rziha HJ, Osterrieder N.
(1996).
Alterations in the equine herpesvirus type-1 (EHV-1) strain RacH during attenuation.
Zentralbl Veterinarmed B, 43(1), 1-14.
https://doi.org/10.1111/j.1439-0450.1996.tb00282.x Publication
Researcher Affiliations
- Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilian University of Munich, Germany.
MeSH Terms
- Animals
- Base Sequence
- Blotting, Northern
- Blotting, Southern
- Cell Line
- Cricetinae
- DNA, Viral / analysis
- DNA, Viral / chemistry
- DNA, Viral / genetics
- Female
- Gene Deletion
- Herpesviridae Infections / immunology
- Herpesviridae Infections / prevention & control
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / genetics
- Herpesvirus 1, Equid / immunology
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / immunology
- Horse Diseases / prevention & control
- Horses
- Kidney / cytology
- Kidney / embryology
- Mesocricetus
- Mice
- Mice, Inbred BALB C
- Molecular Sequence Data
- Phenotype
- Restriction Mapping
- Swine
- Vaccines, Attenuated / genetics
- Vaccines, Attenuated / immunology
Citations
This article has been cited 17 times.- Yang L, Wang M, Cheng A, Yang Q, Wu Y, Huang J, Tian B, Jia R, Liu M, Zhu D, Chen S, Zhao X, Zhang S, Ou X, Mao S, Gao Q, Sun D. Features and Functions of the Conserved Herpesvirus Tegument Protein UL11 and Its Binding Partners. Front Microbiol 2022;13:829754.
- Schnabel CL, Babasyan S, Rollins A, Freer H, Wimer CL, Perkins GA, Raza F, Osterrieder N, Wagner B. An Equine Herpesvirus Type 1 (EHV-1) Ab4 Open Reading Frame 2 Deletion Mutant Provides Immunity and Protection from EHV-1 Infection and Disease. J Virol 2019 Nov 15;93(22).
- Schnabel CL, Wimer CL, Perkins G, Babasyan S, Freer H, Watts C, Rollins A, Osterrieder N, Wagner B. Deletion of the ORF2 gene of the neuropathogenic equine herpesvirus type 1 strain Ab4 reduces virulence while maintaining strong immunogenicity. BMC Vet Res 2018 Aug 22;14(1):245.
- Said A, Elmanzalawy M, Ma G, Damiani AM, Osterrieder N. An equine herpesvirus type 1 (EHV-1) vector expressing Rift Valley fever virus (RVFV) Gn and Gc induces neutralizing antibodies in sheep. Virol J 2017 Aug 14;14(1):154.
- Izume S, Kirisawa R, Ohya K, Ohnuma A, Kimura T, Omatsu T, Katayama Y, Mizutani T, Fukushi H. The full genome sequences of 8 equine herpesvirus type 4 isolates from horses in Japan. J Vet Med Sci 2017 Jan 24;79(1):206-212.
- Słońska A, Cymerys J, Godlewski MM, Dzieciątkowski T, Tucholska A, Chmielewska A, Golke A, Bańbura MW. Equine herpesvirus type 1 (EHV-1)-induced rearrangements of actin filaments in productively infected primary murine neurons. Arch Virol 2014 Jun;159(6):1341-9.
- Ma G, Feineis S, Osterrieder N, Van de Walle GR. Identification and characterization of equine herpesvirus type 1 pUL56 and its role in virus-induced downregulation of major histocompatibility complex class I. J Virol 2012 Apr;86(7):3554-63.
- Goodman LB, Wimer C, Dubovi EJ, Gold C, Wagner B. Immunological correlates of vaccination and infection for equine herpesvirus 1. Clin Vaccine Immunol 2012 Feb;19(2):235-41.
- Ahn BC, Kim S, Zhang Y, Charvat RA, O'Callaghan DJ. The early UL3 gene of equine herpesvirus-1 encodes a tegument protein not essential for replication or virulence in the mouse. Virology 2011 Nov 10;420(1):20-31.
- Kang MH, Banfield BW. Pseudorabies virus tegument protein Us2 recruits the mitogen-activated protein kinase extracellular-regulated kinase (ERK) to membranes through interaction with the ERK common docking domain. J Virol 2010 Sep;84(17):8398-408.
- Rosas C, Van de Walle GR, Metzger SM, Hoelzer K, Dubovi EJ, Kim SG, Parrish CR, Osterrieder N. Evaluation of a vectored equine herpesvirus type 1 (EHV-1) vaccine expressing H3 haemagglutinin in the protection of dogs against canine influenza. Vaccine 2008 May 2;26(19):2335-43.
- Lyman MG, Randall JA, Calton CM, Banfield BW. Localization of ERK/MAP kinase is regulated by the alphaherpesvirus tegument protein Us2. J Virol 2006 Jul;80(14):7159-68.
- Guggemoos S, Just FT, Neubauer A. The equine herpesvirus 1 UL20 product interacts with glycoprotein K and promotes egress of mature particles. J Virol 2006 Jan;80(1):95-107.
- Trapp S, von Einem J, Hofmann H, Köstler J, Wild J, Wagner R, Beer M, Osterrieder N. Potential of equine herpesvirus 1 as a vector for immunization. J Virol 2005 May;79(9):5445-54.
- von Einem J, Wellington J, Whalley JM, Osterrieder K, O'Callaghan DJ, Osterrieder N. The truncated form of glycoprotein gp2 of equine herpesvirus 1 (EHV-1) vaccine strain KyA is not functionally equivalent to full-length gp2 encoded by EHV-1 wild-type strain RacL11. J Virol 2004 Mar;78(6):3003-13.
- Meindl A, Osterrieder N. The equine herpesvirus 1 Us2 homolog encodes a nonessential membrane-associated virion component. J Virol 1999 Apr;73(4):3430-7.
- Osterrieder N, Neubauer A, Brandmüller C, Kaaden OR, O'Callaghan DJ. The equine herpesvirus 1 IR6 protein that colocalizes with nuclear lamins is involved in nucleocapsid egress and migrates from cell to cell independently of virus infection. J Virol 1998 Dec;72(12):9806-17.
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