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Development of immortalized rhesus macaque kidney cells supporting infection with a panel of viruses
Stefanie Reiter1, Sabine Gärtner1, Katharina Decker1
1Infection Biology Unit, German Primate Center, Leibniz Institute for Primate Research, Göttingen, Germany.
Abstract:
Non-human primate (NHP)-based model systems faithfully reproduce various viral diseases including Ebola, influenza, AIDS and Zika. However, only a small number of NHP cell lines are available and generation of additional cell lines could help to refine these models. We immortalized rhesus macaque kidney cells by lentiviral transduction with a vector encoding telomerase reverse transcriptase (TERT) and report the generation of three TERT-immortalized cell lines derived from rhesus macaque kidney. Expression of the kidney podocyte marker podoplanin on these cells was demonstrated by flow cytometry. Quantitative real-time PCR (qRT-PCR) was employed to demonstrate induction of MX1 expression upon stimulation with interferon (IFN) or viral infection, suggesting a functional IFN system. Further, the cell lines were susceptible to entry driven by the glycoproteins of vesicular stomatitis virus, influenza A virus, Ebola virus, Nipah virus and Lassa virus as assessed by infection with retroviral pseudotypes. Finally, these cells supported growth of Zika virus and the primate simplexviruses Cercopithecine alphaherpesvirus 2 and Papiine alphaherpesvirus 2. In summary, we developed IFN-responsive rhesus macaque kidney cell lines that allowed entry driven by diverse viral glycoproteins and were permissive to infection with Zika virus and primate simplexviruses. These cell lines will be useful for efforts to analyze viral infections of the kidney in macaque models.
Insights
Researchers developed new kidney cell lines from rhesus macaques. These interferon-responsive cells support various viral infections, enhancing non-human primate models for diseases like Ebola and Zika.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Non-human primate (NHP) models are crucial for studying viral diseases like Ebola, influenza, AIDS, and Zika.
- Limited availability of NHP cell lines restricts the refinement of these valuable disease models.
Purpose of the Study:
- To generate novel, immortalized kidney cell lines from rhesus macaques.
- To characterize these cell lines for their responsiveness to interferon and susceptibility to viral infections.
Main Methods:
- Immortalization of rhesus macaque kidney cells using lentiviral transduction with telomerase reverse transcriptase (TERT).
- Flow cytometry to assess podoplanin expression (kidney podocyte marker).
- Quantitative real-time PCR (qRT-PCR) to evaluate interferon-stimulated gene (MX1) induction.
- Retroviral pseudotype infections to test viral glycoprotein-mediated entry.
- Zika virus and primate simplexvirus infections to assess permissiveness.
Main Results:
- Successfully generated three TERT-immortalized rhesus macaque kidney cell lines.
- Confirmed expression of the kidney podocyte marker podoplanin.
- Demonstrated functional interferon (IFN) system with induced MX1 expression upon stimulation.
- Confirmed susceptibility to entry by glycoproteins of vesicular stomatitis virus, influenza A virus, Ebola virus, Nipah virus, and Lassa virus.
- Confirmed permissiveness to Zika virus and primate simplexviruses (Cercopithecine alphaherpesvirus 2, Papiine alphaherpesvirus 2).
Conclusions:
- Developed novel, interferon-responsive rhesus macaque kidney cell lines.
- These cell lines facilitate entry of diverse viral glycoproteins and support replication of specific viruses.
- The new cell lines offer a valuable tool for studying viral kidney infections in macaque models.
