title: Bryostatin activates HIV-1 latent expression in human astrocytes through a PKC and NF-ĸB-dependent mechanism.
abstract: Multiple studies have shown that HIV-1 patients may develop virus reservoirs that impede eradication; these reservoirs include the central nervous system (CNS). Despite an undetectable viral load in patients treated with potent antiretrovirals, current therapy is unable to purge the virus from these latent reservoirs. To broaden the inhibitory range and effectiveness of current antiretrovirals, the potential of bryostatin was investigated as a latent HIV-1 activator. We used primary astrocytes, NHA cells, and astrocytoma cells U-87. Infected cells with HIV-1(NL4.3) were treated with bryostatin alone or in combination with different inhibitors. HIV-1 production was quantified by using ELISA. Transcriptional activity was measured using luciferase reporter gene assays by using lipofectin. We performed cotransfection experiments of the LTR promoter with the active NF-κB member p65/relA. To confirm the NF-κB role, Western blot and confocal microscopy were performed. Bryostatin reactivates latent viral infection in the NHA and U87 cells via activation of protein kinase C (PKC)-alpha and -delta, because the PKC inhibitors rottlerin and GF109203X abrogated the bryostatin effect. No alteration in cell proliferation was found. Moreover, bryostatin strongly stimulated LTR transcription by activating the transcription factor NF-κB. Bryostatin could be a beneficial adjunct to the treatment of HIV-1 brain infection.
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Laura
authors.initials: L
authors.lastname: Díaz
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Marta
authors.initials: M
authors.lastname: Martínez-Bonet
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Javier
authors.initials: J
authors.lastname: Sánchez
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Alejandra
authors.initials: A
authors.lastname: Fernández-Pineda
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: José Luis
authors.initials: JL
authors.lastname: Jiménez
authors.affiliation: Department of Cell Biology, Physiology and Immunology, Instituto Maimónides de Investigaciones Biomédicas de Córdoba (IMIBIC)/Reina Sofia University Hospital/University of Córdoba, Córdoba, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Muñoz
authors.affiliation: Departamento de Enfermedades Infecciosas, Hospital Universitario Ramón y Cajal, and IRYCIS.
authors.forename: Santiago
authors.initials: S
authors.lastname: Moreno
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Susana
authors.initials: S
authors.lastname: Álvarez
authors.affiliation: Laboratorio InmunoBiología Molecular, Plataforma de Laboratorio, Hospital General Universitario Gregorio Marañón; Instituto de Investigación Sanitaria Gregorio Marañón, Madrid; Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN).
authors.forename: Ma Ángeles
authors.initials: MÁ
authors.lastname: Muñoz-Fernández
mesh_terms: D001253:Astrocytes / Q000378:metabolism* / Q000821:virology*; D054713:Bryostatins / Q000494:pharmacology*; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral / Q000187:drug effects / Q000235:genetics; D015658:HIV Infections / Q000378:metabolism / Q000821:virology; D016325:HIV Long Terminal Repeat / Q000187:drug effects / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects / Q000235:genetics; D017735:Virus Latency / Q000187:drug effects* / Q000235:genetics
chemical_list: D054713:Bryostatins; D016328:NF-kappa B; C046785:bryostatin 1; D011493:Protein Kinase C