HIV‐1 promonocytic and lymphoid cell lines: an in vitro model of in vivo mitochondrial and apoptotic lesion |
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| Authors: | Constanza Morén Ingrid González‐Casacuberta Carmen Álvarez‐Fernández Maria Bañó Marc Catalán‐Garcia Mariona Guitart‐Mampel Diana Luz Juárez‐Flores Ester Tobías José Milisenda Francesc Cardellach Josep Maria Gatell Sonsoles Sánchez‐Palomino Glòria Garrabou |
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| Affiliation: | 1. Muscle Research and Mitochondrial Function Laboratory, Cellex‐IDIBAPS, Faculty of Medicine‐University of Barcelona, Internal Medicine Department‐Hospital Clínic of Barcelona (HCB), Barcelona, Spain;2. Centro de Investigación Biomédica en Red (CIBER) de Enfermedades Raras (CIBERER), Madrid, Spain;3. Cellex‐IDIBAPS, Faculty of Medicine‐University of Barcelona, Infectious Diseases Unit‐Hospital Clínic of Barcelona (HCB), Barcelona, Spain |
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| Abstract: | To characterize mitochondrial/apoptotic parameters in chronically human immunodeficiency virus (HIV‐1)‐infected promonocytic and lymphoid cells which could be further used as therapeutic targets to test pro‐mitochondrial or anti‐apoptotic strategies as in vitro cell platforms to deal with HIV‐infection. Mitochondrial/apoptotic parameters of U1 promonocytic and ACH2 lymphoid cell lines were compared to those of their uninfected U937 and CEM counterparts. Mitochondrial DNA (mtDNA) was quantified by rt‐PCR while mitochondrial complex IV (CIV) function was measured by spectrophotometry. Mitochondrial‐nuclear encoded subunits II–IV of cytochrome‐c‐oxidase (COXII‐COXIV), respectively, as well as mitochondrial apoptotic events voltage‐dependent‐anion‐channel‐1(VDAC‐1)‐content and caspase‐9 levels] were quantified by western blot, with mitochondrial mass being assessed by spectrophotometry (citrate synthase) and flow cytometry (mitotracker green assay). Mitochondrial membrane potential (JC1‐assay) and advanced apoptotic/necrotic events (AnexinV/propidium iodide) were measured by flow cytometry. Significant mtDNA depletion spanning 57.67% (P < 0.01) was found in the U1 promonocytic cells further reflected by a significant 77.43% decrease of mitochondrial CIV activity (P < 0.01). These changes were not significant for the ACH2 lymphoid cell line. COXII and COXIV subunits as well as VDAC‐1 and caspase‐9 content were sharply decreased in both chronic HIV‐1‐infected promonocytic and lymphoid cell lines (<0.005 in most cases). In addition, U1 and ACH2 cells showed a trend (moderate in case of ACH2), albeit not significant, to lower levels of depolarized mitochondrial membranes. The present in vitro lymphoid and especially promonocytic HIV model show marked mitochondrial lesion but apoptotic resistance phenotype that has been only partially demonstrated in patients. This model may provide a platform for the characterization of HIV‐chronicity, to test novel therapeutic options or to study HIV reservoirs. |
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| Keywords: | apoptosis cell models HIV‐infection HIV progression in vitro modelling mitochondria |
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