Establishment of a cell line with stable expression of mCherry-EGFP tandem fluorescent-tagged LC3B for studying the impact of HIV-1 infection on autophagic flux

Abstract

Increasing evidence indicates that HIV-1 infection has an impact on cell autophagy, and a susceptible cell line is required for studying the relationship of HIV-1 with autophagy. However, there is limited information on the optimal cell line to evaluate the changes of autophagy affected by HIV infection. In this study cell line TZM-tfLC3B was constructed to express mCherry-EGFP tandem fluorescent tagged LC3B (tfLC3B) by stable transfection of tfLC3B as well as allowing X4/R5 tropic HIV-1 replication. The monitoring of autophagic flux in TZM-tfLC3B was achieved by observing fluorescent puncta. HIV-1 virus-like particles lacking replicative nucleic acid could induce autophagy in TZM-tfLC3B in an envelope glycoprotein dependent manner. These data suggest that TZM-tfLC3B will be a useful tool for studying the HIV-1-induced autophagy modulation of host cells.

Keywords: Autophagy; LC3B; Stable transfection; TZM-bl; Virus-like particle.