The persistent infection of a canine thymus cell line by equine infectious anaemia virus and preliminary data on the production of viral antigens

Bouillant, A. M. P.; Nielsen, K.; Ruckerbauer, G M; Samagh, B S; Hare, W.C.D.

doi:10.1016/0166-0934(86)90056-x

Cited by 12 publications

(5 citation statements)

References 17 publications

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“…This adherent epithelial cell line (kindly provided by Prof. L. Sherman) is derived from normal fetal canine thymus and was found to be susceptible to transfection, infection and proliferation by BIV and EIAV [ 41 , 50 ]. These replicating cells were grown in Dulbecco’s modified eagle’s medium (DMEM) supplemented with 10 % heat-inactivated fetal calf serum (FCS), 2% L-glutamine and 1% penicillin-streptomycin antibiotics.…”

Section: Methodsmentioning

confidence: 99%

The dUTPase-related gene of bovine immunodeficiency virus is critical for viral replication, despite the lack of dUTPase activity of the encoded protein

2014

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BackgroundDeoxyuridine 5′-triphosphate nucleotide-hydrolases (dUTPases) are essential for maintaining low intra-cellular dUTP/dTTP ratios. Therefore, many viruses encode this enzyme to prevent dUTP incorporation into their genomes instead of dTTP. Among the lentiviruses, the non-primate viruses express dUTPases. In bovine immunodeficiency virus (BIV), the putative dUTPase protein is only 74 residues-long, compared to ~130 residues in other lentiviruses.ResultsIn this study, the recombinant BIV dUTPase, as well as infectious wild-type (WT) BIV virions, were shown to lack any detectable dUTPase activity. Controls of recombinant dUTPase from equine infectious anemia virus (EIAV) or of EIAV virions showed substantial dUTPase activities. To assess the importance of the dUTPase to BIV replication, we have generated virions of WT BIV or BIV with mutations in the dUTPase gene. The two mutant viral dUTPases were the double mutant D48E/N57S (in the putative enzyme active site and its vicinity) and a deletion of 36 residues. In dividing Cf2Th cells and under conditions where the WT virus was infectious and generated progeny virions, both mutant viruses were defective, as no progeny viruses were generated. Analyses of the integrated viral cDNA showed that cells infected with the mutant virions carry in their genomic DNA levels of integrated BIV DNA that are comparable to those in WT BIV-infected cells.ConclusionsThe herby presented results show that the two BIV mutants with the modified dUTPase gene could infect cells, as viral cDNA was synthesized and integrated into the host cell DNA. However, no virions were generated by cells infected by these mutants. The most likely explanation is that either the integrated cDNA of the mutants is defective (due to potential multiple mutations, introduced during reverse-transcription) or that the original dUTPase mutations have led to severe blocks in viral replication at steps post integration. These results emphasize the importance of the dUTPase-related sequence to BIV replication, despite the lack of any detectable catalytic activity.Electronic supplementary materialThe online version of this article (doi:10.1186/1742-4690-11-60) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

The dUTPase-related gene of bovine immunodeficiency virus is critical for viral replication, despite the lack of dUTPase activity of the encoded protein

2014

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“…Infection can be performed in vitro in various equine cells such as monocyte-derived macrophages [74,102,137], tissue macrophages derived from the spleen [137] or bone marrow [89], endothelial cells [104] or fibroblasts [72,77,99] and in canine and feline transformed cell lines [8,10,58,72].…”

Section: Cell Tropism and Strains Of Virusmentioning

confidence: 99%

Equine Infectious Anemia Virus (EIAV): what has HIV?s country cousin got to tell us?

2004

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-Equine Infectious Anemia Virus (EIAV) is a lentivirus, of the Retrovirus family, with an almost worldwide distribution, infecting equids. It causes a persistent infection characterized by recurring febrile episodes associating viremia, fever, thrombocytopenia, and wasting symptoms. The disease is experimentally reproducible by inoculation of Shetland ponies or horses with EIAV pathogenic strains. Among lentiviruses, EIAV is unique in that, despite a rapid virus replication and antigenic variation, most animals progress from a chronic stage characterized by recurring peaks of viremia and fever to an asymptomatic stage of infection. The inapparent carriers remain infective for life, as demonstrated by experimental transfer of blood to naive animals. The understanding of the correlates of this immune control is of great interest in defining vaccine strategies. Research on EIAV, this "country cousin" of HIV (Human Immunodeficiency Virus), over the last five decades has produced some interesting results on natural immunological control of lentivirus replication and disease and on the nature and role of virus variation in persistence and pathogenesis. These studies are of interest in the context of HIV and efforts to develop a vaccine. This review will focus on some of the most recent results. equine infectious anemia virus / lentivirus / equids / vaccine / viral evolution

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“…Primary eMDM cultures have been used, but have been of relatively limited utility owing to their short-lived nature, to the laborious isolation procedures necessary to establish the cultures, and to the great variability revealed by these primary infections. To facilitate EIAV in vitro studies some authors have adapted primary EIAV strains to growth in equine dermal cells (ED) (Malmquist et al, 1973), fetal equine kidney cells (FEK) (Montelaro et al, 1982), and to the heterologous malignant canine thymus cell line (Cf2Th) (Bouillant et al, 1986). In addition, some EIAV strains have been shown to replicate in equine endothelial cells (Maury et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Characterization of an equine macrophage cell line: Application to studies of EIAV infection

Fidalgo-Carvalho¹,

Craigo

Barnes

et al. 2009

Veterinary Microbiology

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EIAV is a monocyte/macrophage tropic virus. To date, even though EIAV has been under investigation for numerous years, very few details have been elucidated about EIAV/macrophage interactions. This is largely due to the absence of an equine macrophage cell line that would support viral replication. Herein we describe the spontaneous immortalization and generation of a clonal equine macrophage-like (EML) cell line with the functional and immunophenotype characteristics of differentiated equine monocyte derived macrophage(s) (eMDM(s)). These cells possess strong non-specific esterase (NSE) activity, are able to phagocytose fluorescent bioparticles, and produce nitrites in response to LPS. The EML-3C cell line expresses the EIAV receptor for cellular entry (ELR1) and supports replication of the virulent EIAV PV biological clone. Thus, EML-3C cells provide a useful cell line possessing equine macrophage related properties for the growth and study of EIAV infection as well as of other equine macrophage tropic viruses.

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The persistent infection of a canine thymus cell line by equine infectious anaemia virus and preliminary data on the production of viral antigens

Cited by 12 publications

References 17 publications

The dUTPase-related gene of bovine immunodeficiency virus is critical for viral replication, despite the lack of dUTPase activity of the encoded protein

The dUTPase-related gene of bovine immunodeficiency virus is critical for viral replication, despite the lack of dUTPase activity of the encoded protein

Equine Infectious Anemia Virus (EIAV): what has HIV?s country cousin got to tell us?

Characterization of an equine macrophage cell line: Application to studies of EIAV infection

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