title: Depletion of mitochondrial DNA copies/cell in peripheral blood mononuclear cells in HIV-1-infected treatment-naïve patients.
abstract: OBJECTIVE
  Mitochondrial toxicity is believed to be the main reason for adverse effects related to nucleoside reverse transcriptase inhibitors (NRTIs). The aim of the present study was to compare mitochondrial toxicity in NRTI-treated HIV-positive patients, HIV-positive treatment-naïve patients and HIV-negative controls by comparing mitochondrial DNA (mtDNA) copies/cell in peripheral blood mononuclear cells (PBMCs) and lactate/pyruvate (L/P) ratios in the different groups.
  
  
  METHODS
  We enrolled 60 participants in the study: 31 patients on combined antiretroviral therapy (CART), 14 HIV-positive treatment-naive patients and 15 HIV-negative controls. mtDNA (copies/cell) in peripheral blood was analysed using quantitative real-time polymerase chain reaction (PCR). Standard curves and serial dilutions of plasmid-cloned mitochondrion and retinoblastoma (RB1) PCR products with known concentrations were generated to estimate the mtDNA and nuclear DNA (nDNA) copy numbers in each sample. The L/P ratio was enzymatically and spectrophotometrically analysed in samples from individuals in a fasted, non-exercise state. Results The median mtDNA copy number was 63 copies/cell (interquartile range 33-94) in HIV-positive patients and 153 (132-283) in HIV-negative controls (P<0.001). No significant difference was seen between the HIV-positive NRTI-exposed patients and the HIV-positive treatment-naive patients. Current use of didanosine was negatively correlated with depletion of mtDNA (r=-0.36, P=0.046). HIV-positive patients also had a higher L/P ratio compared with HIV-negative controls (P=0.004).
  
  
  CONCLUSIONS
  The number of mtDNA copies/cell in PBMCs was depleted in HIV-positive treatment-naive patients as well as in HIV-positive NRTI-exposed patients. HIV-positive patients also had a higher L/P ratio compared with HIV-negative controls, which supports this conclusion. The study suggests that neither mtDNA in PBMCs nor L/P ratio is a good marker of NRTI-associated mitochondrial toxicity.
authors.affiliation: Department of Infectious Diseases, Ullevaal University Hospital, Faculty Division, University of Oslo, Oslo, Norway. anne.maagard@medisin.uio.no
authors.forename: A
authors.initials: A
authors.lastname: Maagaard
authors.forename: M
authors.initials: M
authors.lastname: Holberg-Petersen
authors.forename: E A
authors.initials: EA
authors.lastname: Kvittingen
authors.forename: L
authors.initials: L
authors.lastname: Sandvik
authors.forename: J N
authors.initials: JN
authors.lastname: Bruun
mesh_terms: D000328:Adult; D000368:Aged; D019380:Anti-HIV Agents / Q000009:adverse effects; D015415:Biomarkers / Q000097:blood; D004272:DNA, Mitochondrial / Q000097:blood* / Q000187:drug effects; D016903:Drug Monitoring / Q000379:methods; D005260:Female; D015658:HIV Infections / Q000097:blood* / Q000188:drug therapy; D015497:HIV-1 / Q000302:isolation & purification; D006801:Humans; D019344:Lactic Acid / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000737:chemistry*; D008297:Male; D008875:Middle Aged; D019289:Pyruvic Acid / Q000097:blood; D012367:RNA, Viral / Q000097:blood; D018894:Reverse Transcriptase Inhibitors / Q000009:adverse effects; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D019380:Anti-HIV Agents; D015415:Biomarkers; D004272:DNA, Mitochondrial; D012367:RNA, Viral; D018894:Reverse Transcriptase Inhibitors; D019344:Lactic Acid; D019289:Pyruvic Acid