Dejiang Zhou1, Stephen A Spector. 1. Department of Pediatrics, Division of Infectious Diseases, University of California San Diego, La Jolla, California, 92093-0672, USA.
Abstract
OBJECTIVES: Human immunodeficiency virus type-1 (HIV-1) induces a series of alterations in the host cell that modify the intracellular environment in favor of viral replication, survival and spread. This research examined the impact of HIV-1 infection on autophagy in HIV-1 infected cells. METHODS: Protein extracts of HIV-1 infected and control CD4+ T-lymphocytes and U937 cells were semi-quantified by western blot. The autophagy-related protein Beclin 1, a Bcl-2 associated protein, and the 16 kD microtubule-associated protein (MAP) light chain three (LC3) which is essential for autophagy were quantified and validated using the intracellular protein GAPDH as an internal standard. Beclin 1 mRNA was quantified by real-time reverse transcriptase-polymerase chain reaction. Autophagosomes were assessed by visualization under confocal microscopy following intracellular staining of the LC3 protein. RESULTS: Following infection of human peripheral blood CD4+ T-cells or U937 cells with HIV-1 for 48 h, the autophagy protein Beclin 1 and LC3 II, which is essential for autophagy, were found to be markedly decreased. Beclin 1 mRNA expression was also reduced. Autophagosomes were reduced in HIV-1-infected cells. The reduction of autophagic protein expression and autophagosomes in HIV-1-infected cells could be overcome by amino acid starvation or rapamycin. CONCLUSIONS: These data demonstrate that HIV-1 infection can down-regulate autophagy in infected cells during acute infection, and provide new insights into HIV-1-induced cell death and disease-related pathogenesis.
OBJECTIVES:Humanimmunodeficiency virus type-1 (HIV-1) induces a series of alterations in the host cell that modify the intracellular environment in favor of viral replication, survival and spread. This research examined the impact of HIV-1 infection on autophagy in HIV-1 infected cells. METHODS: Protein extracts of HIV-1 infected and control CD4+ T-lymphocytes and U937 cells were semi-quantified by western blot. The autophagy-related protein Beclin 1, a Bcl-2 associated protein, and the 16 kD microtubule-associated protein (MAP) light chain three (LC3) which is essential for autophagy were quantified and validated using the intracellular protein GAPDH as an internal standard. Beclin 1 mRNA was quantified by real-time reverse transcriptase-polymerase chain reaction. Autophagosomes were assessed by visualization under confocal microscopy following intracellular staining of the LC3 protein. RESULTS: Following infection of human peripheral blood CD4+ T-cells or U937 cells with HIV-1 for 48 h, the autophagy protein Beclin 1 and LC3 II, which is essential for autophagy, were found to be markedly decreased. Beclin 1 mRNA expression was also reduced. Autophagosomes were reduced in HIV-1-infected cells. The reduction of autophagic protein expression and autophagosomes in HIV-1-infected cells could be overcome by amino acid starvation or rapamycin. CONCLUSIONS: These data demonstrate that HIV-1 infection can down-regulate autophagy in infected cells during acute infection, and provide new insights into HIV-1-induced cell death and disease-related pathogenesis.
Authors: J L Rossio; M T Esser; K Suryanarayana; D K Schneider; J W Bess; G M Vasquez; T A Wiltrout; E Chertova; M K Grimes; Q Sattentau; L O Arthur; L E Henderson; J D Lifson Journal: J Virol Date: 1998-10 Impact factor: 5.103
Authors: R P van Gijlswijk; H J Zijlmans; J Wiegant; M N Bobrow; T J Erickson; K E Adler; H J Tanke; A K Raap Journal: J Histochem Cytochem Date: 1997-03 Impact factor: 2.479
Authors: William T Jackson; Thomas H Giddings; Matthew P Taylor; Sara Mulinyawe; Marlene Rabinovitch; Ron R Kopito; Karla Kirkegaard Journal: PLoS Biol Date: 2005-04-26 Impact factor: 8.029