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Analysis of Simian Immunodeficiency Virus-specific CD8+ T-cells in Rhesus Macaques by Peptide-MHC-I Tetramer Staining
Published on: December 23, 2016
Extensively deleted simian immunodeficiency virus (SIV) DNA in macaques inoculated with supercoiled plasmid DNA
1Unité de Pathogénie des Infections à Lentivirus, INSERM U372, Parc Scientifique et Technologique de Luminy, 13273 Marseille, France.
Abstract:
Using long-distance DNA PCR, we prospectively followed rhesus monkeys that had been inoculated intramuscularly with supercoiled plasmid DNA encoding intact simian immunodeficiency virus (SIV). From 4 to 10 weeks postinoculation onward, we detected extensively deleted proviral genomes along with full-length viral genomes in peripheral blood mononuclear cells (PBMC) in adult macaques. During their chronic asymptomatic phase of infection, the frequency of deleted proviral genomes was similar in PBMC and lymph nodes. The latter, however, harbored significantly more full-length proviral DNA than PBMC, consistent with the lack of effective antiviral cytotoxic T-cell activity in lymph nodes described by others during human immunodeficiency virus infection. After the macaques progressed to AIDS, full-length proviral DNA became equally abundant in lymph nodes and in PBMC. We have demonstrated that although a single molecular species of proviral DNA was inoculated, genomic diversity was detected within a short time, thus confirming the genetic instability of the SIV genome in vivo.
Insights
Simian immunodeficiency virus (SIV) infection in rhesus monkeys shows rapid genomic diversity, with deleted SIV DNA appearing quickly. This genetic instability highlights challenges in controlling SIV and related viruses.
Area of Science:
- Virology
- Immunology
- Genetics
Background:
- Simian immunodeficiency virus (SIV) infection in macaques serves as a model for human immunodeficiency virus (HIV) infection.
- Understanding viral genome dynamics is crucial for developing effective antiviral strategies.
- Previous research suggests differences in immune responses between peripheral blood and lymph nodes during viral infections.
Purpose of the Study:
- To investigate the in vivo genetic instability and diversity of the SIV genome after inoculation.
- To compare the distribution of full-length and deleted SIV proviral DNA in peripheral blood mononuclear cells (PBMC) and lymph nodes over time.
- To correlate viral genome dynamics with disease progression and immune activity in macaques.
Main Methods:
- Prospective follow-up of rhesus monkeys inoculated with SIV plasmid DNA.
- Long-distance DNA PCR to detect and quantify proviral DNA.
- Analysis of SIV genome integrity (full-length vs. deleted) in PBMC and lymph nodes.
- Monitoring disease progression to AIDS.
Main Results:
- Extensively deleted SIV proviral genomes were detected alongside full-length genomes in PBMC from 4 to 10 weeks post-inoculation.
- During the chronic phase, deleted SIV DNA was equally frequent in PBMC and lymph nodes, but lymph nodes had more full-length SIV DNA.
- In the AIDS phase, full-length SIV DNA became equally abundant in both tissues.
- Genomic diversity of SIV was observed rapidly, confirming in vivo genetic instability.
Conclusions:
- The SIV genome exhibits significant genetic instability in vivo, leading to rapid diversification.
- Tissue-specific differences in SIV proviral DNA distribution exist, potentially influenced by local immune activity.
- These findings provide insights into viral persistence and evolution in SIV/HIV infection.
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