Stavudine Reduces NLRP3 Inflammasome Activation and Modulates Amyloid-β Autophagy

J Alzheimers Dis. 2019;72(2):401-412. doi: 10.3233/JAD-181259.


Background: Alzheimer's disease (AD) is associated with the accumulation of amyloid-β (Aβ) within senile plaques in the brain and neuroinflammation, possibly driven by the activation of the NLRP3 inflammasome. Nucleoside reverse transcriptase inhibitors (NRTI) hamper the NLRP3 inflammasome assembly.

Objective: We utilized an in vitro model reproducing the Aβ-driven inflammation seen in AD to analyze whether stavudine (D4T), a prototypical NRTI, modulates Aβ-mediated inflammasome activation and the ability of macrophages to eliminate Aβ via phagocytosis and autophagy.

Methods: THP-1-derived macrophages were stimulated in vitro with Aβ42 or with Aβ42 after LPS-priming in the presence/absence of D4T. NLRP3 and TREM2 expression was analyzed by RT-PCR; phagocytosis, as well as ASC-Speck formation, was analyzed by Amnis FlowSight Imaging; NLRP3-produced cytokines were quantified by ELISA and, finally, autophagy was analyzed by measuring p-ERK1/2, p-AKT, beclin, p70-S6Kinase, and Lamp by ELISA and western blot.

Results: IL-1β, IL-18, and caspase-1 were increased whereas Aβ phagocytosis and TREM2 were reduced in LPS+Aβ42-stimulated cells. D4T reduced NLRP3 assembly as well as IL-18 and caspase-1 production, but did not affect IL-1β production and TREM2 expression. Notably, whereas D4T reduced Aβ phagocytosis, Aβ autophagy by macrophages was stimulated by D4T, as witnessed by the down-modulation of ERK1/2 and AKT phosphorylation and the upregulation of beclin, LAMP, and p70-S6K, their downstream targets.

Conclusion: In this in vitro model of AD, D4T reduces NLRP3 inflammasome-associated inflammation and stimulates Aβ autophagy by macrophages. It will be interesting to verify the possibly beneficial effects of D4T in the clinical scenario.

Keywords: Alzheimer’s disease; D4T; NLRP3-inflammasome; amyloid-β phagocytosis; autophagy; cytokines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid beta-Peptides / antagonists & inhibitors*
  • Amyloid beta-Peptides / pharmacology*
  • Autophagy / drug effects*
  • Cells, Cultured
  • Cytokines / metabolism
  • Enzyme Activation / drug effects
  • Gene Expression Regulation / drug effects
  • Humans
  • Inflammasomes / drug effects*
  • MAP Kinase Signaling System / drug effects
  • Macrophages / drug effects
  • Membrane Glycoproteins / antagonists & inhibitors
  • Membrane Glycoproteins / biosynthesis
  • NLR Family, Pyrin Domain-Containing 3 Protein / antagonists & inhibitors*
  • Phagocytosis / drug effects
  • Receptors, Immunologic / antagonists & inhibitors
  • Receptors, Immunologic / biosynthesis
  • Reverse Transcriptase Inhibitors / pharmacology*
  • Stavudine / pharmacology*


  • Amyloid beta-Peptides
  • Cytokines
  • Inflammasomes
  • Membrane Glycoproteins
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP3 protein, human
  • Receptors, Immunologic
  • Reverse Transcriptase Inhibitors
  • TREM2 protein, human
  • Stavudine