Quantification of mitochondrial toxicity in HIV-infected individuals by quantitative PCR compared to flow cytometry

Cytometry B Clin Cytom. 2013 Jan-Feb;84(1):55-8. doi: 10.1002/cyto.b.21045. Epub 2012 Oct 8.

Abstract

Background: Non-invasive diagnostic assays to evaluate mitochondrial toxicity could have significant clinical utility for HIV-infected individuals on antiretroviral therapy (ART).

Methods: This study compared the ratio of mitochondrial to nuclear DNA determined by quantitative polymerase chain reaction (qPCR) to the ratio of mitochondrial to nuclear-encoded proteins by flow cytometry, in peripheral blood mononuclear cells from 73 HIV-infected individuals with and without risk factors for mitochondrial toxicity.

Results: PCR detected similar mitochondrial/nuclear DNA in HIV-infected individuals without a history of ART, and those receiving ART with lipodystrophy, lipoatrophy, or a history of suspected lactic acidosis. However, the ratio was significantly greater in ART-untreated compared to those receiving either stavudine or didanosine. In contrast, flow cytometry did not detect any differences in mitochondrial/nuclear protein (Lin et al., Cytometry B 2009;76B:181-190). There was no correlation between the assays (rho = -0.05, P = 0.65).

Conclusions: Assessment of the mitochondrial/nuclear DNA ratio by qPCR performed better than the mitochondrial/nuclear-encoded protein ratio by flow cytometry to detect adverse effects of nucleoside analogs on mitochondria.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acidosis, Lactic
  • Adult
  • Anti-HIV Agents / therapeutic use
  • Antiretroviral Therapy, Highly Active
  • DNA, Mitochondrial / analysis*
  • Didanosine / therapeutic use
  • Female
  • Flow Cytometry
  • HIV Infections / blood
  • HIV Infections / diagnosis*
  • Humans
  • Leukocytes, Mononuclear / cytology
  • Lipodystrophy
  • Male
  • Middle Aged
  • Mitochondria / pathology*
  • Mitochondrial Proteins / analysis*
  • Real-Time Polymerase Chain Reaction
  • Stavudine / therapeutic use

Substances

  • Anti-HIV Agents
  • DNA, Mitochondrial
  • Mitochondrial Proteins
  • Stavudine
  • Didanosine