HIV-1 infected Cell Pellets and Biotinylated Cannabidiol LC-MS/MS

Previous studies have shown that cannabis use in people living with HIV-1 is associated with a lower viral load, lower circulating CD16+ monocytes, and high CD4+ T-cell count, suggesting a potential therapeutic application. HIV-1 infected U1 monocytes and primary macrophages were used to assess the effects of CBD in HIV-1 infection. Our data suggests that CBD treatment results in a significant reduction in the number of EVs released from infected cells. Additional findings suggest that this may be mediated by a reduction in viral transcription and autophagy activation. To see the effect of CBD in autophagy activation, U1 and U1 Monocyte Derived Macrophages cells were cultured and lysed. D-Biotin or Biotinylated CBD pull-down from the lysates was performed using Streptavidin-Sepharose beads, which were washed with TNE50 / TNE300 (high salt) + 0.1% NP40 buffers. The complexes were analyzed by mass spectrometry. Reactome enrichment analysis was used to assess the CBD pull-down of HIV-1 infected monocytes (U1) and HIV-1 infected MDM proteins for involvement of the autophagy pathway.