RESUMO
Placental inflammations (villitis) are accompanied by loss of the syncytiotrophoblast, which is the cellular barrier separating maternal blood from fetal tissue in the villous placenta. We propose that syncytiotrophoblast loss is mediated by adhesion of activated maternal monocytes. This hypothesis was tested with a co-culture model of peripheral blood monocytes and placental syncytiotrophoblasts. We find that LPS-activated monocytes adhere to interferon-gamma (IFN-gamma)-treated syncytiotrophoblasts via monocyte LFA-1 for >48 h, during which time the monocytes induce trophoblast apoptosis and subsequent damage of the trophoblast layer. Optimal monocyte-mediated syncytiotrophoblast death requires both lipopolysaccharide (LPS) and IFN-gamma and is inhibited by either anti-tumor necrosis factor (TNF) antibody or epidermal growth factor. Syncytiotrophoblast damage is largely limited to culture surfaces in the vicinity of bound monocytes. These results show that activated maternal monocytes bound to the placental barrier can induce focal damage mediated by the inflammatory cytokine TNF-alpha and suggest a route for maternal leukocyte infiltration into the fetal stroma.
Assuntos
Apoptose/fisiologia , Vilosidades Coriônicas/patologia , Antígeno-1 Associado à Função Linfocitária/metabolismo , Monócitos/citologia , Trofoblastos/citologia , Fator de Necrose Tumoral alfa/fisiologia , Anticorpos Monoclonais/farmacologia , Adesão Celular , Fator de Crescimento Epidérmico/farmacologia , Feminino , Células Gigantes , Humanos , Inflamação , Interferon gama/farmacologia , Lipopolissacarídeos/farmacologia , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Gravidez , Células Estromais/fisiologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Whether cell-free human immunodeficiency virus type 1 (HIV-1) can productively infect placental trophoblasts (which in turn could transmit the virus into the fetal circulation) is controversial but essential to know for the evaluation of alternative routes (such as cell-mediated infection or trophoblast damage). We have addressed infection factors such as cell purity, source, culture methods, and activation states as well as virus variant and detection methods to conclusively determine the outcome of trophoblast challenge by free virus. Pure (> 99.98%) populations of trophoblasts from 11 different placentas were challenged at a multiplicity of infection (MOI) as high as 6 with five different HIV-1 variants, three of which are non-syncytium-forming, macrophage-tropic isolates from infected infants, with and without coinfection with cytomegalovirus; these preparations were monitored for productive infection for up to 3 weeks after challenge by five different criteria, the most sensitive of which were cocultivation with target cells that can detect virus at an MOI of 10(-7) and HIV DNA PCR that detects 30 virus copies per 10(5) cells. Infection was never detected. However, molecularly cloned T-cell (pNL4-3)- and macrophage (pNLAD8)-tropic provirus plasmids, when transfected into primary trophoblasts, yielded productive infections, indicating that trophoblasts do not suppress late-stage virus replication and assembly. Because of the purity of the trophoblast preparations, the extended length of the infection culture period, the number of trophoblast preparations and virus types examined, the sensitivity of the bioassays and molecular detection assays, and the observations that trophoblasts can support virus replication from provirus, the results of this study strongly argue that free virus cannot infect primary villous trophoblasts.
Assuntos
Citomegalovirus/fisiologia , HIV-1/fisiologia , Provírus/fisiologia , Trofoblastos/virologia , Membrana Celular/virologia , Células Cultivadas , Técnicas de Cocultura , Citoplasma/virologia , DNA Viral/metabolismo , Variação Genética , HIV-1/isolamento & purificação , Células HeLa , Humanos , Lactente , Macrófagos/virologia , Provírus/genética , Recombinação Genética , Linfócitos T/virologia , Transfecção , Trofoblastos/citologia , Replicação ViralRESUMO
In the placenta, as in other organs, the development and maintenance of the differentiated phenotype depend on a balance between cell proliferation, maturation, and death. We are interested in the mechanisms that regulate the survival and differentiation of placental trophoblasts and have recently demonstrated that the inflammatory cytokines tumor necrosis factor alpha (TNF alpha) and gamma interferon (IFN gamma) act in concert to induce apoptotic cell death in normal cytotrophoblasts in culture. In this report we show that exposure to epidermal growth factor (EGF), a 6,700 dalton polypeptide that is abundantly expressed in maternal and fetal tissues, blocks the in vitro TNF/IFN-induced cytotoxicity of human cytotrophoblasts and syncytiotrophoblasts from normal term placentas. This antagonistic effect is dose-related (10-10 M EGF, half-maximal) and proceeds via the interruption of an early step in the cytokine-induced apoptotic response. These observations suggest a novel role for EGF in normal placental development and indicate that the interplay between EGF, TNF alpha, and IFN gamma may determine the rate of trophoblast growth and renewal during gestation.
Assuntos
Apoptose/efeitos dos fármacos , Fator de Crescimento Epidérmico/farmacologia , Fatores de Crescimento de Fibroblastos , Interferon gama/farmacologia , Trofoblastos/citologia , Fator de Necrose Tumoral alfa/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas/citologia , Células Cultivadas/efeitos dos fármacos , Fator 10 de Crescimento de Fibroblastos , Fator 7 de Crescimento de Fibroblastos , Citometria de Fluxo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Substâncias de Crescimento/farmacologia , Humanos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Trofoblastos/efeitos dos fármacosRESUMO
The placental syncytiotrophoblast (ST) is a terminally differentiated epithelial cell monolayer that constitutes the outermost boundary between fetal and maternal tissues and performs a variety of synthetic, secretory, and transport functions essential for the maintenance of pregnancy. Although it is known that the ST arises from the underlying germinal layer of mononuclear cytotrophoblasts (Langhans' cells) by a process of cell fusion, the molecular mechanisms involved in this process are unclear. In order to address this question, we have investigated the effects of macrophage colony-stimulating factor (CSF-1) and granulocyte-macrophage colony-stimulating factor (GM-CSF), lymphohemopoietic cytokines implicated in mammalian placental development, on the in vitro morphological and functional differentiation of human trophoblast. Both CSF-1 and GM-CSF stimulated cytotrophoblast aggregation into large multinucleated structures composed of extensive patches of syncytium interspersed with mononuclear cells. Concomitant with this morphological differentiation was upregulation of the production of the placental hormones placental lactogen and chorionic gonadotrophin. Placental fibroblasts derived from the villous stroma that underlies the trophoblastic epithelium were found to produce both GM-CSF and CSF-1 under the control of the trophoblast-derived cytokines IL-1 and TNF alpha. These observations suggest that a network of interrelated cytokines operates within the basal (fetal) aspects of the villous stroma where they are situated to play a significant role in the morphological and functional development of the human placenta.
Assuntos
Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Fator Estimulador de Colônias de Macrófagos/farmacologia , Trofoblastos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Gonadotropina Coriônica/metabolismo , Citocinas/farmacologia , Interações Medicamentosas , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Interleucina-1/farmacologia , Lactogênio Placentário/metabolismo , Receptor de Fator Estimulador de Colônias de Macrófagos/biossíntese , Trofoblastos/citologia , Trofoblastos/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Regulação para CimaRESUMO
Mononuclear phagocytes (MP) are probably the most capable effector cells of the body in the defense against virulent strains of E. histolytica. Killing of E. histolytica by MP appears to involve both oxidative and non-oxidative mechanisms. Thus, in this study we have investigated whether trophozoites of an axenic virulent strain E. histolytica HM1:IMSS (EH) were capable of eliciting an oxidative response in pure populations of freshly isolated human monocytes. Using a luminol-enhanced chemiluminescence assay we demonstrate that these cells produce a strong respiratory burst when challenged with live amebas over a wide range of MP:EH ratios. Furthermore, pre-incubation of monocytes with recombinant Macrophage Colony Stimulating Factor (M-CSF) could further increase the oxidative metabolism of MP in response to E. histolytica. Our results indicate that, in contrast to what occurs with polymorphonuclear leukocytes, the interaction of E. histolytica with MP leads to the production of reactive oxygen intermediates by this cells. The enhancement of this potent microbicidal mechanism by inflammatory cytokines may further increase the amebicidal capacity of human mononuclear phagocytes.
Assuntos
Entamoeba histolytica , Fator Estimulador de Colônias de Macrófagos/farmacologia , Monócitos/efeitos dos fármacos , Explosão Respiratória/efeitos dos fármacos , Adulto , Animais , Entamoeba histolytica/patogenicidade , Humanos , Medições Luminescentes , Espécies Reativas de Oxigênio/metabolismo , VirulênciaRESUMO
The cytokines are multifunctional polypeptide hormones, produced by a variety of cells, that participate in the regulation of many biological processes. Essentially acting as intercellular messengers, they play a central role in the maintenance of homeostasis in normal tissues. Cytokines are key mediators of both local and systemic immune-inflammatory responses; therefore, disturbances in their secretion, response and/or regulation have been implicated in the pathophysiology of several conditions in which an exaggerated auto-destructive component appears to play a role. Conversely, deficits in cytokine production probably impair the host's ability to mount an effective immune response and may underlie the increased susceptibility to microbial infection observed, for example, in the neonate or in the malnourished patient. A review of the principal aspects of the biology of cytokines is presented together with the evidence supporting their involvement in several pathological states and the potential therapeutic applications of these mediators as novel immunomodulating agents.