Monitoring Reactivation of Latent HIV by Label-Free Gradient Light Interference Microscopy

Abstract

SummaryLatent human immunodeficiency virus (HIV) reservoirs in infected individuals present the largest barrier to a cure. The first step towards overcoming this challenge is to understand the science behind latency-reactivation interplay. Fluorescence imaging of GFP-tagged HIV has been the main method for studying reactivation of latent HIV in individually infected cells. In this paper, we report insights provided by label-free, gradient light interference microscopy (GLIM) about the changes in measures including dry mass, diameter, and dry mass density associated with infected cells that occur upon reactivation. We discovered that mean cell dry mass and mean diameter of latently infected cells treated with reactivating drug, TNF-α, are higher for cells with reactivated HIV as compared to those with latent disease. Results also indicate that cells with mean dry mass and diameter less than 10pg and 8µm, respectively, remain exclusively in the latent state. Also, cells with mean dry mass greater than 23pg and mean diameter greater than 11µm have a higher probability of reactivating. This study is significant as it presents a new label-free approach to quantify latent reactivation of a virus in single cells based on changes in cell morphology.