Establishment and characterization of fetal equine kidney and lung cells with extended lifespan. Susceptibility to equine gammaherpesvirus infection and transfection efficiency.
Abstract: Due to the slow growth of equine gammaherpesviruses, isolation of these viruses requires cells that can be propagated long term and show clear cytopathy following infection. Equine cell lines with extended lifespan were established from primary cells originating from equine fetal kidney and lung by transfecting the cells with the retroviral vector LXSN116E6E7 containing the human papilloma virus oncogenes 16 E6 and E7. The transfected equine kidney cell line and equine lung cell line can be propagated for more than 40 passages, whereas the corresponding primary cells only for 10-12 passages. The primary cells and the derived cell lines can be infected with equine gammaherpesvirus 2 (EHV-2) with similar efficiency. However EHV-5 can be grown to a substantially higher titer in the kidney cell line than their primary counterpart, with cytopathic effect visible three days earlier than in the primary cells. Due to rapid cell growth the lung cell line is difficult to use for virus production. The kidney cell line was four times more susceptible to transfection as compared to the primary kidney cells. On the other hand no difference was between the lung cell line and the primary lung cells in transfection efficiency. The cell lines can be a valuable tool for investigating gammaherpesviruses, and possibly other viruses infecting horses.
Publication Date: 2016-05-12 PubMed ID: 27173610DOI: 10.1007/s11626-016-0046-9Google Scholar: Lookup
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Summary
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This research article discusses the development and analysis of long-lasting equine kidney and lung cells to study equine gammaherpesvirus infection and its effect on transfection efficiency.
Extended Lifespan of Equine Cells
The researchers generated equine cells with a prolonged lifespan from primary equine fetal kidney and lung cells. The process involved:
- Transfection of the cells with a retroviral vector (LXSN116E6E7) which contains human papilloma virus oncogenes 16 E6 and E7.
- This technique extended the propagation lifespan of the transfected equine kidney and lung cell lines beyond 40 passages. Notably, the primary cells could be propagated only for 10-12 passages.
Susceptibility to Equine Gammaherpesvirus
The article also discusses the susceptibility of these cells to equine gammaherpesvirus, specifically the EHV-2 strain. The key findings include:
- The primary cells and their derived cell lines were infected with EHV-2 with similar efficiency.
- The kidney cell line could be infected by EHV-5 to a significantly higher level than its primary cell counterpart, indicating higher susceptibility.
- The infected kidney cells showed cytopathic effects three days earlier than the primary cells.
- However, the lung cell line was not ideal for virus production due to its rapid growth.
Transfection Efficiency
The research assesses the efficiency at which these cells could be transfected with foreign DNA. It reveals that:
- The kidney cell line was four times more susceptible to transfection than the primary kidney cells.
- Interestingly, no significant difference was observed between the lung cell line and primary lung cells in terms of transfection efficiency.
Potential Applications
The researchers suggest that these new equine cell lines can be a valuable tool for studying gammaherpesviruses, and potentially, other viruses that infect horses. By enabling an extended cell lifespan and high transfection efficiency, this development could significantly enhance research into equine virology.
Cite This Article
APA
Thorsteinsdóttir L, Torsteinsdóttir S, Svansson V.
(2016).
Establishment and characterization of fetal equine kidney and lung cells with extended lifespan. Susceptibility to equine gammaherpesvirus infection and transfection efficiency.
In Vitro Cell Dev Biol Anim, 52(8), 872-877.
https://doi.org/10.1007/s11626-016-0046-9 Publication
Researcher Affiliations
- Institute for Experimental Pathology, University of Iceland, Keldur, Keldnavegur 3, 112, Reykjavík, Iceland. liljatho@hi.is.
- Institute for Experimental Pathology, University of Iceland, Keldur, Keldnavegur 3, 112, Reykjavík, Iceland.
- Institute for Experimental Pathology, University of Iceland, Keldur, Keldnavegur 3, 112, Reykjavík, Iceland.
MeSH Terms
- Animals
- Cell Line / virology
- Disease Susceptibility
- Gammaherpesvirinae / pathogenicity
- Herpesviridae Infections / pathology
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Horses
- Humans
- Kidney / cytology
- Kidney / virology
- Lung / cytology
- Lung / virology
- Transfection
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Citations
This article has been cited 2 times.- Thorsteinsdóttir L, Guðmundsson GÖ, Jensson H, Torsteinsdóttir S, Svansson V. Isolation of equid alphaherpesvirus 3 from a horse in Iceland with equine coital exanthema. Acta Vet Scand 2021 Feb 2;63(1):6.
- Thorsteinsdóttir L, Jónsdóttir S, Stefánsdóttir SB, Andrésdóttir V, Wagner B, Marti E, Torsteinsdóttir S, Svansson V. The effect of maternal immunity on the equine gammaherpesvirus type 2 and 5 viral load and antibody response. PLoS One 2019;14(6):e0218576.
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