In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry.

ABSTRACT: We demonstrate for the first time in-cell dynamic nuclear polarization (DNP) in conjunction with flow cytometry sorting to address the cellular heterogeneity of in-cell samples. Utilizing a green fluorescent protein (GFP) reporter of HIV reactivation, we correlate increased ¹⁵N resonance intensity with cytokine-driven HIV reactivation in a human cell line model of HIV latency. As few as 10% GFP+ cells could be detected by DNP nuclear magnetic resonance (NMR). The inclusion of flow cytometric sorting of GFP+ cells prior to analysis by DNP-NMR further boosted signal detection through increased cellular homogeneity with respect to GFP expression. As few as 3.6 million ¹⁵N-labeled GFP+ cells could be readily detected with DNP-NMR. Importantly, cell sorting allowed for the

SUBMITTER: Overall SA 

PROVIDER: S-EPMC7369949 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature