In vivo effects of highly active antiretroviral therapies containing the protease inhibitor nelfinavir on mitochondrially driven apoptosis.
BACKGROUND
In vitro studies have reported controversial effects of protease inhibitors (PIs) on mitochondrially driven apoptosis. Additionally, since PIs in the clinical setting are almost always given in combination with nucleoside analogues, which may have negative effects on mitochondrial DNA (mtDNA), the impact of PI-containing highly active antiretroviral therapy (HAART) on apoptosis and mtDNA content is unclear.


METHODS
A cross-sectional study was performed including 20 HIV-negative (HIV-) patients, 16 HIV-positive, antiretroviral-naive (HIV+) patients and 17 HIV-positive patients receiving the PI nelfinavir (NFV) plus zidovudine and lamivudine (AZT+3TC) or didanosine and stavudine (ddl+d4T)--collectively known as HIV+PI--as first-line antiretroviral treatment for at least 12 months. Peripheral blood mononuclear cells (PBMCs) were isolated. BCL2 expression (anti-apoptotic) and the levels of the cleaved, active form of caspase-9 (pro-apoptotic) were determined by western blot. An index of mitochondrially driven apoptotic activation was estimated calculating the ratio caspase-9:BCL2. Mitochorldrial DNA content was measured by real-time PCR.


RESULTS
BCL2 expression was lower in HIV+ than in HIV-patients (P < 0.01), whereas levels of caspase-9 were higher (P = 0.001). The caspase-9:BCL2 ratio was significantly increased in HIV+ compared with HIV-individuals (P < 0.001). Mitochondrial DNA content was also decreased in HIV+ compared with HIV-patients (P < 0.001). The HIV+PI group exhibited a trend to normalization for BCL2 expression and caspase-9 compared with the HIV+ group, whereas the caspase-9:BCL2 ratio significantly improved (decreased, P < 0.05 compared with HIV+ group). The mtDNA content in the HIV+PI group was similar to that of the HIV+ group, although the results of mtDNA content differed depending on whether NFV was combined with AZT+3TC (preserved) or with ddl+d4T (depleted). Conversely, no differences were found in apoptotic markers between the two subgroups of HIV+PI.


CONCLUSIONS
NFV-based PI-containing HAART regimens may exert some beneficial effects counteracting the increased mitochondrially driven apoptosis present in HIV-infected people.
Miró
Oscar
O
Mitochondrial Research Laboratory, Hospital Clinic, IDIBAPS, Barcelona, Catalonia, Spain. omiro@clinic.ub.es
Villarroya
Joan
J
Garrabou
Glòria
G
López
Sònia
S
Rodríguez de la Concepción
Marisa
M
Pedrol
Enric
E
Martínez
Esteban
E
Giralt
Marta
M
Gatell
Josep M
JM
Cardellach
Francesc
F
Casademont
Jordi
J
Villarroya
Francesc
F
D000328:Adult; D019380:Anti-HIV Agents / Q000627:therapeutic use*; D023241:Antiretroviral Therapy, Highly Active; D017209:Apoptosis / Q000187:drug effects*; D003430:Cross-Sectional Studies; D005260:Female; D015658:HIV Infections / Q000188:drug therapy* / Q000276:immunology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000502:physiology*; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D019888:Nelfinavir / Q000494:pharmacology* / Q000627:therapeutic use; D011480:Protease Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use
D019380:Anti-HIV Agents; D011480:Protease Inhibitors; D019888:Nelfinavir