Quantifying integrated SIV-DNA by repetitive-sampling Alu-gag PCR

Area of Science:

• Virology
• Immunology
• Molecular Biology

Background:

• Antiretroviral therapy (ART) suppresses HIV-1 but does not eliminate the latent viral reservoir.
• Accurate measurement of the latent reservoir (LR) is essential for developing and assessing HIV cure strategies.
• Simian immunodeficiency virus (SIV)/simian-human immunodeficiency virus (SHIV) models in rhesus macaques (RMs) are critical for preclinical research.

Discussion:

• This study optimized an ex vivo repetitive sampling Alu-gag PCR assay to quantify integrated SIV-DNA in ART-naïve and ART-experienced RMs.
• The assay demonstrated reliable detection and quantification of integrated SIV-DNA in various tissues and cell types.
• Results showed correlations between integrated and total SIV-DNA levels, providing a comprehensive view of viral persistence.

Key Insights:

• Integrated SIV-DNA levels were quantified in ART-naïve RMs during acute and chronic SHIV infection.
• In ART-treated RMs, integrated SIV-DNA was detected in lymph nodes and spleen, with comparable total and integrated levels after prolonged ART.
• Peripheral blood CD4+ T cells from ART-treated RMs showed similar levels of total and integrated SIV-DNA, positively correlated.

Outlook:

• The validated assay serves as a critical tool for interventional studies targeting HIV/SIV cure.
• Measuring integrated SIV-DNA aids in estimating the size of the latent reservoir in RMs.
• This work advances the development of strategies to reduce viral persistence and achieve HIV/SIV remission.