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Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids.
Abstract: Equine infectious anemia virus (EIAV) infection of horses is characterized by recurring cycles of disease and viremia that typically progress to an inapparent infection in which clinical symptoms are absent as host immune responses maintain control of virus replication indefinitely. The dynamics of EIAV viremia and its association with disease cycles have been well characterized, but there has been to date no comprehensive quantitative analyses of the specific tissue sites of EIAV infection and replication in experimentally infected equids during acute disease episodes and during asymptomatic infections in long-term inapparent carriers. To characterize the in vivo site(s) of viral infection and replication, we developed a quantitative competitive PCR assay capable of detecting 10 copies of viral DNA and a quantitative competitive reverse transcription-PCR assay with a sensitivity of about 30 copies of viral singly spliced mRNA. Animals were experimentally infected with one of two reference viruses: the animal-passaged field isolate designated EIAV(Wyo) and the virulent cell-adapted strain designated EIAV(PV). Tissues and blood cells were isolated during the initial acute disease or from asymptomatic animals and analyzed for viral DNA and RNA levels by the respective quantitative assays. The results of these experiments demonstrated that the appearance of clinical symptoms in experimentally infected equids coincided with rapid widespread seeding of viral infection and replication in a variety of tissues. During acute disease, the predominant cellular site of viral infection and replication was the spleen, which typically accounted for over 90% of the cellular viral burden. In asymptomatic animals, viral DNA and RNA persisted in virtually all tissues tested, but at extremely low levels, a finding indicative of tight but incomplete immune control of EIAV replication. During all disease states, peripheral blood mononuclear cells (PBMC) were found to harbor less than 1% of the cellular viral burden. These quantitative studies demonstrate that tissues, rather than PBMC, constitute the predominant sites of virus replication during acute disease in infected equids and serve as resilient reservoirs of virus infection, even in the presence of highly effective immune responses that maintain a stringent control of virus replication in long-term inapparent carriers. Thus, these observations with EIAV, a predominantly macrophage-tropic lentivirus, highlight the role of tissues in sequestering lentiviral infections from host immune surveillance.
Publication Date: 2000-03-09 PubMed ID: 10708426PubMed Central: PMC111810DOI: 10.1128/jvi.74.7.3112-3121.2000Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This study investigates the specific sites in horses’ bodies where the Equine infectious anemia virus (EIAV) thrives and replicates during both acute outbreaks and periods of asymptomatic infection. The researchers used advanced methods to detect and quantify the virus in various tissues and blood cells, identifying the spleen as the primary site of infection and replication during acute disease while virtually all tissues harbored low virus levels in asymptomatic animals.
Objective and Methodology
- The primary aim of the research was to identify the explicit tissue sites of EIAV infection and replication as this was overlooked in prior research. The researchers sought clarity on these areas to understand the virus better and potentially develop targeted treatment methods.
- Two distinct quantitative assays were established by the researchers for the study, a quantitative competitive PCR assay and a quantitative competitive reverse transcription-PCR assay. These assays were created to detect the presence and quantity of viral DNA and RNA.
- The subjects of the experiment, equids (horses and alike), were infected with two different versions of the virus for the experiment, namely EIAV(Wyo) and EIAV(PV).
- Various tissues and blood cells were then isolated from the animals during the periods of acute disease and asymptomatic illness. These samples were analyzed for viral DNA and RNA levels using the two quantitative assays developed.
Key Findings
- During the phase of acute disease, the research found that the virus efficiently spread and replicated in various tissues. The spleen was identified as the most common site, accounting for over 90% of the viral burden in cells. This indicates that the spleen is a primary site for EIAV infection and replication during periods of acute disease.
- In the case of asymptomatic animals, the researchers found that the virus’ DNA and RNA persisted in practically all examined tissues, though at extremely low levels, suggesting a strong but not entirely perfect immune control of EIAV replication.
- The study emphasized that tissues serve as the dominant sites for virus replication during acute disease and seemingly resilient reservoirs of virus infection, even amidst highly effective immune responses. In contrast, the role of peripheral blood mononuclear cells (PBMCs) in harboring the virus was determined to be minimal, accounting for less than 1% of the cellular viral burden.
Conclusion and Implications
- The research, therefore, deduced that, similar to other lentiviruses that primarily target macrophages, EIAV uses tissues to hide from the host’s immune responses. This highlights the significant role tissues play in sustaining lentiviral infections despite the host’s robust immune reaction.
- The evidence generated through this study can potentially guide future efforts towards the development of more targeted treatment strategies for EIAV. By understanding where the virus hides and thrives in the host body, scientists and clinicians can develop methods to specifically target these areas, enhancing the effectiveness of treatment options.
Cite This Article
APA
Harrold SM, Cook SJ, Cook RF, Rushlow KE, Issel CJ, Montelaro RC.
(2000).
Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids.
J Virol, 74(7), 3112-3121.
https://doi.org/10.1128/jvi.74.7.3112-3121.2000 Publication
Researcher Affiliations
- Department of Molecular Genetics and Biochemistry, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
MeSH Terms
- Animals
- Base Sequence
- DNA Primers
- DNA, Viral / analysis
- Equine Infectious Anemia / virology
- Horses
- Infectious Anemia Virus, Equine / genetics
- Infectious Anemia Virus, Equine / physiology
- RNA Splicing
- RNA, Viral / analysis
- RNA, Viral / genetics
- Reverse Transcriptase Polymerase Chain Reaction
- Virus Replication
Grant Funding
- R01 CA049296 / NCI NIH HHS
- T32 AI007487 / NIAID NIH HHS
- R01 CA49296 / NCI NIH HHS
- 5T32 AI07487 / NIAID NIH HHS
- P41 RR006009 / NCRR NIH HHS
- 2 P41 RR06009 / NCRR NIH HHS
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