RESUMEN
Levels of serum ferritin are increased in AIDS patients in relation to the progression of the disease. To establish whether or not this in vivo increase could be due to a direct effect of the virus on the infected cells, three HIV-permissive cell lines, the CD4-positive HeLa-T4-6c and C8166 cells and the CD4-negative RD cells, were infected with HIV-1 strains. The expression of ferritin was followed during the course of acute infection, in parallel to other cellular components. Unexpectedly, all three cell lines showed a phase of decrease in their ferritin content after infection by HIV-1, not justified by the modest and late increase of ferritin in the fluids, due to disruption of infected cells. Since ferritin is involved in the control of cell growth and DNA synthesis, its downregulation may be implied both in cell toxicity and DNA abnormalities due to HIV infection.
Asunto(s)
Ferritinas/metabolismo , VIH-1/fisiología , Antígenos CD4/metabolismo , Regulación hacia Abajo , Células HeLa , Humanos , Células Tumorales Cultivadas , Replicación Viral , Microglobulina beta-2/metabolismoRESUMEN
We asked whether HIV-1 had the capacity to establish a persistent infection of cultured human diploid fibroblasts. Human strains of normal diploid embryo lung fibroblasts were infected with HIV-1 of the HTLV-IIIB and HIV-1P1 strains. Infection was followed over time, to analyze HIV expression. Virus production (intra- and extracellular virus) was evaluated as follows: ability to form syncytia in the C8166 T cell line, production of p24 and other viral antigens (ELISA and indirect immunofluorescence), search for a gag sequence in cell DNA by the polymerase chain reaction followed by hybridization to an HIV-1-specific probe (SK19). Cell-free culture supernatant was used as a virus source to infect de novo fibroblasts and C8166 T cells. Infection of cultured fibroblasts with either the HTLV-IIIB or HIV-1P1 strain led regularly to the establishment of persistently infected cultures. Fibroblast cells were capable of continuous virus production for at least 10 months. The released virus was capable of reinfecting cultured fibroblasts and of producing cytopathic effects in the C8166 T cell line. However, when compared to wild-type strains, the infectious virus derived from fibroblasts showed a prolonged replication cycle and a decreased ability to form syncytia in the T cell line. Therefore, HIV-1 can establish a persistent and productive infection in normal lung fibroblasts. The data are consistent with the hypothesis that in vivo, at least in the lung, fibroblasts may represent a virus reservoir and that infection of these cells may lead to the production of attenuated variants of HIV.
Asunto(s)
VIH-1/fisiología , VIH-1/patogenicidad , Replicación Viral , Línea Celular , Embrión de Mamíferos , Ensayo de Inmunoadsorción Enzimática , Fibroblastos/virología , Células Gigantes , Antígenos VIH/análisis , Antígenos VIH/biosíntesis , Proteína p24 del Núcleo del VIH/análisis , Proteína p24 del Núcleo del VIH/biosíntesis , Humanos , Pulmón , Fusión de Membrana , Reacción en Cadena de la Polimerasa , Linfocitos TAsunto(s)
Infecciones por VIH/inmunología , VIH-1/inmunología , Antígenos HLA-DR/biosíntesis , Proteínas Portadoras/metabolismo , Línea Celular , Citocinas/farmacología , Fibroblastos/inmunología , Fibroblastos/virología , Infecciones por VIH/virología , Humanos , Receptores de Hialuranos , Técnicas In Vitro , Molécula 1 de Adhesión Intercelular/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores Mensajeros de Linfocitos/metabolismoRESUMEN
The production of cytokines by HIV-infected cells from adherent tissues as well as their effects on HIV replication in the same cells were investigated. CD4-transfected HeLa-T4-6c epithelial cells, CD4-positive normal lung fibroblasts and CD4-negative RD rhabdomyosarcoma cells were infected with HIV-1. All cultures were permissive for virus replication, which was completed within 48-72 h by Hela-T4-6c and RD cells and 2-3 weeks in normal fibroblasts. During the course of HIV replication, a series of cytokines (particularly IL-6 and TNF alpha) was produced and released in parallel to the peak of virus growth, in amounts varying with the cell system studied. Treatment of cultures with recombinant cytokines given at concentrations in the range of those induced by HIV-1 indicated that IL-6 and TNF alpha caused an increase of: i) CD4 expression, ii) HIV absorption to uninfected cells, and iii) release of infectious virions by infected cells. The fact that HIV-1 absorption and spread can be mediated by HIV-induced cytokines may be relevant in the pathogenesis of the in vivo disease, as it may constitute a possible self-enhancing model of HIV infection also in the solid tissues.