HIGH SALT CONCENTRATIONS POTENTIATES RE- VERSAL OF HIV-1 LATENCY THROUGH AN NF-ĸB-DE- PENDENT MECHANISM

MELUCCI GANZARAIN, CLAUDIA; GERBER, PEHUÉN PEREYRA; MAZZITELLI, I.; BLEICHMAR, LUCÍA; ERRA DIAZ, FERNANDO; GEFFNER, JORGE

Virtual

Congreso; Reunión Conjunta SAIC. SAI. AAFE. NANOMED-AR 2021; 2021

BACKGROUND. The HIV-1 reservoir comprises a pool of infected CD4 T cells in which the expression of the viral genome remains latent, constituting the main barrier to a sterilizing cure. The ‘shock& kill’ strategy involves reactivating HIV-1 transcription in these cells to facilitate their elimination. Candidate Latency Reversal Agents (LRAs), however, have failed in clinical trials. There is, therefore, an unmet need to develop new approaches aimed to reactivate viral transcription. Considering that high salt concentrations are a common finding in secondary lymphatic organs, here we study its effect on HIV-1 latency.METHODS. In this study, we exploit a Jurkat cell line latently infect-ed with an HIV-1 clone containing GFP in place of the viral gene nef (J-Lat). We expose J-Lat cells to control or hypernatremic media (+50 mM NaCl) for 4 h before treatment with LRAs. HIV-1 reactivation, monitored by GFP, is measured by flow cytometry after 24 h. To evaluate the role of p38 and NFAT5 we silence the expression of the proteins with shRNAs. To measure the activity of NF-ĸB we use a reporter cell line together with the degradation of IKBα by western blot.RESULTS. First, we find that transient exposure to hypernatremic media sensitises J-Lat cells to HIV-1 latency reversal by two LRAs (PMA: 5,2 ± 2,4% vs. 17,7 ± 4,7% and TNF-α: 10,2 ± 2,6% vs. 30,7 ± 10,0%; control vs. hypernatremic media, n = 4, p < 0,001). Then, we confirm this effect is independent of the activity of MAPK p38 and the transcription factor NFAT5. Finally, we analyze the transcriptional activity of NF-ĸB and observe a significant (p