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An overwhelming obstacle to HIV cure is the presence of long-lived viral reservoirs that can reignite viral infection after removal of combination antiretroviral therapy cART. Yet, our knowledge of the mechanisms of reservoir maintenance and persistence remains incompletely understood. Therefore, in this study, we generated a novel virus model for evaluating individual reactivation events following cART interruption to better understand key aspects of reservoir biology in vivo.

Next-generation sequencing was used to evaluate the number of genetic variants in the stock and in plasma before and after treatment. In the viral stock, individual barcodes, or clonotypes, were identified. During acute infection, an average of barcodes were identified in each of the 10 animals. Between 87 and distinct clonotypes were detected in plasma at peak rebound viraemia in animals from study 1 and between 4 and 7 clonotypes in animals from study 2.

Because the growth rate of each clonotype is equivalent once the virus reaches systemic infection, the measured relative proportions of each clonotype at rebound reflect the time between when each clonotype achieved systemic infection.

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