Recombinant herpesviruses as tools for the study of herpesvirus biology

Kúdelová, M; Belvončíková, Petra; Halásová, Zuzana; Košovský, Ján; Lapuníková, B.; Pančík, P; Režuchová, Ingeborg; Šupolíková, Miroslava; Zelnik, V.

doi:10.4149/av_2013_02_149

Cited by 4 publications

(2 citation statements)

References 77 publications

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“…Genetic modifications of CMV genomes are most conveniently performed using bacterial artificial chromosome (BAC) recombineering [35,36]. Unfortunately, the only existing RhCMV BAC was based on the 68-1 RhCMV strain, an extensively fibroblasts passaged isolate which, compared to circulating and low passage isolates, has acquired a large inversion in the region homologous to one end of the HCMV "unique long" (U L ) sequence of the genome (commonly referred to as the ULb' region), flanked by deletions of multiple ORFs on either side of the inversion [23,37,38].…”

Section: Construction Of a Full Length (Fl) Rhcmv Bac And In Vitro Chmentioning

confidence: 99%

In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome

et al. 2020

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Cytomegaloviruses (CMVs) are highly adapted to their host species resulting in strict species specificity. Hence, in vivo examination of all aspects of CMV biology employs animal models using host-specific CMVs. Infection of rhesus macaques (RM) with rhesus CMV (RhCMV) has been established as a representative model for infection of humans with HCMV due to the close evolutionary relationships of both host and virus. However, the only available RhCMV clone that permits genetic modifications is based on the 68–1 strain which has been passaged in fibroblasts for decades resulting in multiple genomic changes due to tissue culture adaptations. As a result, 68–1 displays reduced viremia in RhCMV-naïve animals and limited shedding compared to non-clonal, low passage isolates. To overcome this limitation, we used sequence information from primary RhCMV isolates to construct a full-length (FL) RhCMV by repairing all mutations affecting open reading frames (ORFs) in the 68–1 bacterial artificial chromosome (BAC). Inoculation of adult, immunocompetent, RhCMV-naïve RM with the reconstituted virus resulted in significant viremia in the blood similar to primary isolates of RhCMV and furthermore led to high viral genome copy numbers in many tissues at day 14 post infection. In contrast, viral dissemination was greatly reduced upon deletion of genes also lacking in 68–1. Transcriptome analysis of infected tissues further revealed that chemokine-like genes deleted in 68–1 are among the most highly expressed viral transcripts both in vitro and in vivo consistent with an important immunomodulatory function of the respective proteins. We conclude that FL-RhCMV displays in vitro and in vivo characteristics of a wildtype virus while being amenable to genetic modifications through BAC recombineering techniques.

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Section: Construction Of a Full Length (Fl) Rhcmv Bac And In Vitro Chmentioning

confidence: 99%

In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome

et al. 2020

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“…There have been presented only a few data on further analysis and characterization of OvHV-1 and its genome (Kúdelová et al, 2013). De described some properties of the JS-3 OvHV-1 isolate, at that time identified as bovid herpesvirus 4.…”

Section: Introductionmentioning

confidence: 99%

Ovine herpesvirus 1 (OVHV-1) thymidine kinase locus sequence analysis: evidence that OVHV-1 belongs to the Macavirus genus of the Gammaherpesvirinae subfamily

Ziak¹,

Koptidesová²,

Ovečková³

et al. 2014

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Summary. -The HindIII-HincII fragment of the 5.5 kbp H11 HindIII clone of ovine herpesvirus 1 (OvHV-1) was cloned and its primary structure was determined by preparation of nested deletion subclones and their sequencing. Sequence analysis of the overlapping clones revealed that 3239 bp OvHV-1 fragment contains complete thymidine kinase (TK) gene, a partial open reading frame of ORF20 and that encoding glycoprotein H (gH). The conserved OvHV-1 TK displayed the highest similarity to homologous TK proteins encoded by members of the Macavirus genus of the Gammaherpesvirinae subfamily. These data including our previous analysis of the partial sequence of VP23 homologue might serve as further evidence that OvHV-1 should be categorized within the genus Macavirus of the Herpesviridae family.Keywords: ovine herpesvirus 1; thymidine kinase; Macavirus; Gammaherpesvirinae * Corresponding author. E-mail: viruzelo@savba.sk; phone: +421-2-59302463. Abbreviation: CPE = cytopathic effect; FUDR = 5΄-fluoro-2΄-deoxyuridine; HSV-1 = herpes simplex virus 1; JSRV = Jaagsiekte sheep retrovirus; MuHV-4 = murid herpesvirus 4; OvHV-1 = ovine herpesvirus 1; SPA = sheep pulmonary adenomatosis; TK = thymidine kinase

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Domestic cats seropositive for Felis catus gammaherpesvirus 1 are often qPCR negative

et al. 2016

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Felis catus gammaherpesvirus 1 (FcaGHV1) is a newly described virus that infects domestic cats. To identify FcaGHV1 antigens, we developed an immunofluorescent antibody assay by expressing FcaGHV1 open reading frames (ORFs) in feline cells and incubating fixed cells with sera from FcaGHV1-positive cats. Of the seven ORFs tested, ORF52 and ORF38 had the strongest, most consistent antibody responses. We used recombinant ORF52 and ORF38 proteins to develop two FcaGHV1 ELISAs. These assays were used to detect reactivity in cats previously tested by qPCR for FcaGHV1 in blood cell DNA. Results indicated 32%FcaGHV1seroprevalence, compared to 15%qPCR-evaluated prevalence (n=133);all but one qPCR positive animal was seropositive. ELISA results confirmed infection risk factors previously identified by qPCR: geographic location, male sex, and adult age. These data suggest that FcaGHV1is a common infection of domestic cats that has a seropositive but often qPCR negative state characteristic of herpesviral latency.

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Recombinant herpesviruses as tools for the study of herpesvirus biology

Cited by 4 publications

References 77 publications

In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome

In vitro and in vivo characterization of a recombinant rhesus cytomegalovirus containing a complete genome

Ovine herpesvirus 1 (OVHV-1) thymidine kinase locus sequence analysis: evidence that OVHV-1 belongs to the Macavirus genus of the Gammaherpesvirinae subfamily

Domestic cats seropositive for Felis catus gammaherpesvirus 1 are often qPCR negative

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