Methods: PBMC were cultured to generate monocyte-derived macrophages (MDM). MDM were treated 24 or 48h with 5ng/ml Tenofovir and/or 109ng/ml Emtricitabine and lysates collected. Primary human astrocytes were treated 24h with 10ng HIV Nef or 5ng/ml Tenofovir + 109ng/ml Emtricitabine + 14ng/ml Raltegravir (ART) and lysates collected. Lysates were analyzed by western blot for LC3-II and p62 (autophagy markers) using Image Studio.
Results: LC3-II levels increased 1.5, 1.6 and 1.7-fold in MDM treated 24h with Tenofovir, Emtricitabine or Truvada, respectively, and p62 level decreased 25% after 24h Truvada, relative to untreated MDM. After 48h Truvada, LC3-II and p62 levels decreased 30% relative to control MDM. This indicates an initial upregulation followed by rapid downregulation of autophagy in antiretroviral-treated MDM. In astrocytes treated with Nef, LC3-II and p62 flux (degradation) increased 3-fold and 1.7-fold respectively, indicating abnormal enhancement of autophagy. Interestingly, while ART treatment increased LC3-II flux 3-fold, p62 degradation decreased 50% relative to controls, signifying a change in autophagy that impacts LC3-II and p62 differently.
Conclusion: Our initial study demonstrates that HIV and commonly prescribed antiretrovirals induce aberrant autophagy dynamics in macrophages and astrocytes. To our knowledge, this is the first time a change in autophagy due to ART has been described in these cells. HIV and ART may disrupt the homeostasis provided by autophagy, contributing to accelerated aging and HAND in HIV-infected people.
A. M. Cuervo, None
J. W. Berman, None