Endotoxin treatment of equine infectious anaemia virus-infected horse macrophage cultures decreases production of infectious virus.
Abstract: Lentiviruses replicate in cells of the immune system, and activation of immune cells has been shown to modulate virus replication. To determine the effects of macrophage activation on replication of equine infectious anaemia virus (EIAV), primary horse macrophage cultures (HMCs) were established from 20 different horses, infected with an avirulent strain of EIAV, and stimulated with 5 microg/ml of bacterial endotoxin. Supernatants collected from HMCs were assayed for the presence of tumour necrosis factor (TNF-alpha) and for production of infectious virus. Results indicated that EIAV replication in vitro varied significantly (P < or = 0.0001) from horse to horse, regardless of the treatment of HMCs. Also, EIAV replication was significantly (P < or = 0.0001) decreased in HMCs stimulated with bacterial endotoxin as compared to untreated HMCs. No significant correlation was found between virus replication and production of TNF-alpha following treatment of virus-infected cells with bacterial endotoxin. However, when HMCs were treated with endotoxin prior to virus infection, inhibition of EIAV replication was proportional to increasing levels of endotoxin. PCR and RT-PCR were used to amplify EIAV proviral DNA and mRNA sequences, respectively, at various time-points following infection. The results indicated that the early events of EIAV replication, up to and including transcription of multiple-spliced mRNAs, were not inhibited by treatment of EIAV-infected macrophages with bacterial endotoxin. This suggests that endotoxin treatment inhibits a posttranscriptional step in the virus replication cycle.
Publication Date: 1998-05-06 PubMed ID: 9568970DOI: 10.1099/0022-1317-79-4-747Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This research investigates how the activation of immune cells through endotoxin treatment can impact the replication of the equine infectious anaemia virus (EIAV) in horse macrophages.
Study Design and Procedure
- The researchers used primary horse macrophage cultures (HMCs) derived from 20 different horses as their study sample.
- These cultures were infected with an avirulent strain of EIAV and then stimulated using 5 microg/ml of bacterial endotoxin.
- They collected supernatants from these cultures to test for the presence of tumour necrosis factor (TNF-alpha) and for the production of infectious virus.
Key Findings
- The study found that EIAV replication varied significantly from horse to horse, irrespective of the HMCs’ treatment, suggesting a degree of individual variability in response.
- It also found that when the HMCs were treated with bacterial endotoxin, there was a significant decrease in EIAV replication compared to untreated HMCs.
- The researchers found no significant correlation between virus replication and the production of TNF-alpha after bacterial endotoxin treatment.
- When the researchers treated the HMCs with endotoxin prior to infecting them with the virus, they found that the inhibition of EIAV replication was proportional to the levels of endotoxin, meaning that a higher endotoxin level resulted in a greater inhibition.
Analysis using PCR and RT-PCR
- The researchers used Polymerase chain reaction (PCR) and Reverse transcription polymerase chain reaction (RT-PCR) methods to amplify EIAV proviral DNA and mRNA sequences at different times following infection.
- The findings from this part of the study suggested that the early stages of EIAV replication, up to and including the transcription of multiple-spliced mRNAs, were not inhibited by treating EIAV-infected macrophages with bacterial endotoxin.
- This pointed towards the conclusion that the endotoxin treatment inhibits a stage of the virus replication cycle that occurs after transcription.
Cite This Article
APA
Smith TA, Davis E, Carpenter S.
(1998).
Endotoxin treatment of equine infectious anaemia virus-infected horse macrophage cultures decreases production of infectious virus.
J Gen Virol, 79 ( Pt 4), 747-755.
https://doi.org/10.1099/0022-1317-79-4-747 Publication
Researcher Affiliations
- Department of Microbiology, Immunology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
MeSH Terms
- Animals
- Base Sequence
- Cells, Cultured
- DNA Primers / genetics
- Endotoxins / pharmacology
- Horses
- Infectious Anemia Virus, Equine / drug effects
- Infectious Anemia Virus, Equine / genetics
- Infectious Anemia Virus, Equine / physiology
- Macrophage Activation
- Macrophages / drug effects
- Macrophages / immunology
- Macrophages / virology
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- RNA, Viral / genetics
- RNA, Viral / metabolism
- Transcription, Genetic / drug effects
- Tumor Necrosis Factor-alpha / biosynthesis
- Virus Replication / drug effects
- Virus Replication / genetics
- Virus Replication / immunology
Grant Funding
- AI30025 / NIAID NIH HHS
Citations
This article has been cited 5 times.- de Pablo-Maiso L, Doménech A, Echeverría I, Gómez-Arrebola C, de Andrés D, Rosati S, Gómez-Lucia E, Reina R. Prospects in Innate Immune Responses as Potential Control Strategies against Non-Primate Lentiviruses. Viruses 2018 Aug 17;10(8).
- Sponseller BA, Clark SK, Gilbertie J, Wong DM, Hepworth K, Wiechert S, Chandramani P, Sponseller BT, Alcott CJ, Bellaire B, Petersen AC, Jones DE. Macrophage effector responses of horses are influenced by expression of CD154. Vet Immunol Immunopathol 2016 Nov 1;180:40-44.
- Tallmadge RL, Brindley MA, Salmans J, Mealey RH, Maury W, Carpenter S. Development and characterization of an equine infectious anemia virus Env-pseudotyped reporter virus. Clin Vaccine Immunol 2008 Jul;15(7):1138-40.
- Belshan M, Park GS, Bilodeau P, Stoltzfus CM, Carpenter S. Binding of equine infectious anemia virus rev to an exon splicing enhancer mediates alternative splicing and nuclear export of viral mRNAs. Mol Cell Biol 2000 May;20(10):3550-7.
- Hansen R, Czub S, Werder E, Herold J, Gosztonyi G, Gelderblom H, Schimmer S, Mazgareanu S, ter Meulen V, Czub M. Abundant defective viral particles budding from microglia in the course of retroviral spongiform encephalopathy. J Virol 2000 Feb;74(4):1775-80.
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