Establishment of a Novel Equine Cell Line for Isolation and Propagation of Equine Herpesviruses

Maeda, Ken; Yasumoto, Shigeru; Tsuruda, Akari; Andoh, Kiyohiko; Kai, K.; Otoi, Takeshige; Matsumura, Tomio

doi:10.1292/jvms.69.989

Cited by 22 publications

(18 citation statements)

References 5 publications

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“…In some cases, cells were co-treated with 100 M N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-fmk; MP Biomedicals, Eschwege, Germany). Fetal liver stromal cell lines were prepared as described previously (22,23). Briefly, fetal liver stromal cells derived from wild-type or anamorsin-deficient E14.5 murine embryos were inoculated onto 0.1% gelatincoated plastic dishes and allowed to proliferate for 2 days.…”

Section: Methodsmentioning

confidence: 99%

Anamorsin, a Novel Caspase-3 Substrate in Neurodegeneration

Yun

Lee

Kim

et al. 2014

Journal of Biological Chemistry

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Background: It is crucial to identify caspase-3 substrates and their roles during neurodegeneration. Results: Our approach identified 46 novel substrates. Furthermore, we found that anamorsin was cleaved by caspase-3 and mapped its cleavage site. Conclusion: Anamorsin may play an antiapoptotic role in the central nervous system. Significance: Our findings contribute to understanding the molecular mechanism underlying role for anamorsin in caspase-3-mediated cell death.

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Section: Methodsmentioning

confidence: 99%

Anamorsin, a Novel Caspase-3 Substrate in Neurodegeneration

Yun

Lee

Kim

et al. 2014

Journal of Biological Chemistry

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“…EHV-1 can readily be propagated in many cell lines, including primary equine cells and cell lines derived from other species (75). In contrast, EHV-4 appears to be restricted mainly to cells derived from horses, such as FHK and NBL-6 cells, or replicates only poorly in very few other cell lines, such as Vero cells (44). To test the hypothesis that gD is a major determinant of EHV-1 and EHV-4 tropism, gD mutant viruses were used to explore the viruses' ability to infect different cell lines, using the same infectious doses (MOI, 0.1), which allowed us to define the host range of each virus in vitro.…”

Section: Generation and Genotypic Characterization Of Mutant Virusesmentioning

confidence: 99%

Glycoproteins D of Equine Herpesvirus Type 1 (EHV-1) and EHV-4 Determine Cellular Tropism Independently of Integrins

Azab

Osterrieder

2012

J Virol

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Equine herpesvirus type 1 (EHV-1) and EHV-4 are genetically and antigenically very similar, but their pathogenic potentials are strikingly different. The differences in pathogenicity between both viruses seem to be reflected in cellular host range: EHV-1 can readily be propagated in many cell types of multiple species, while EHV-4 entry and replication appear to be restricted mainly to equine cells. The clear difference in cellular tropism may well be associated with differences in the gene products involved in virus entry and/or spread from cell to cell. Here we show that (i) most of the EHV-1 permissive cell lines became resistant to EHV-1 expressing EHV-4 glycoprotein D (gD4) and the opposite was observed for EHV-4 harboring EHV-1 gD (gD1). (ii) The absence of integrins did not inhibit entry into and replication of EHV-1 in CHO-K1 or peripheral blood mononuclear cells (PBMC). Furthermore, integrin-negative K562 cells did not acquire the ability to bind to gD1 when ␣V␤3 integrin was overexpressed. (iii) PBMC could be infected with similar efficiencies by both EHV-1 and EHV-4 in vitro. (iv) In contrast to results for equine fibroblasts and cells of endothelial or epithelial origin, we were unable to block entry of EHV-1 or EHV-4 into PBMC with antibodies directed against major histocompatibility complex class I (MHC-I), a result that indicates that these viruses utilize a different receptor(s) to infect PBMC. Cumulatively, we provide evidence that efficient EHV-1 and EHV-4 entry is dependent mainly on gD, which can bind to multiple cell surface receptors, and that gD has a defining role with respect to cellular host range of EHV-1 and EHV-4.

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“…FHK-Tcl3.1 (Andoh et al, 2009;Maeda et al, 2007), 293T, Crandel-Ree feline kidney (CRFK) and cloned porcine kidney (CPK) cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM) (Invitrogen, CA, USA) supplemented with 10% heatinactivated fetal calf serum (FCS), 100 units of penicillin and 100 g/mL streptomycin (Invitrogen, CA, USA). Cells were grown at 37 • C in 5% CO 2 .…”

Section: Cellsmentioning

confidence: 99%

“…Notably, EHV-1 can grow in some non-equine cells, whereas the growth of EHV-4 appears to be restricted to equine cells. We recently established an equine cell line (FHK-Tcl3.1) that is able to host both viruses, and thus permit a more direct comparison of their biology (Andoh et al, 2009;Maeda et al, 2007).…”

Section: Introductionmentioning

confidence: 99%