RESUMEN
The contribution of placental immune responses to congenital Zika virus (ZIKV) syndrome remains poorly understood. Here, we leveraged a mouse model of ZIKV infection to identify mechanisms of innate immune restriction exclusively in the fetal compartment of the placenta. ZIKV principally infected mononuclear trophoblasts in the junctional zone, which was limited by mitochondrial antiviral-signaling protein (MAVS) and type I interferon (IFN) signaling mechanisms. Single nuclear RNA sequencing revealed MAVS-dependent expression of IFN-stimulated genes (ISGs) in spongiotrophoblasts but not in other placental cells that use alternate pathways to induce ISGs. ZIKV infection of Ifnar1-/- or Mavs-/- placentas was associated with greater infection of the adjacent immunocompetent decidua, and heterozygous Mavs+/- or Ifnar1+/- dams carrying immunodeficient fetuses sustained greater maternal viremia and tissue infection than dams carrying wild-type fetuses. Thus, MAVS-IFN signaling in the fetus restricts ZIKV infection in junctional zone trophoblasts, which modulates dissemination and outcome for both the fetus and the pregnant mother.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales , Decidua , Feto , Interferón Tipo I , Placenta , Receptor de Interferón alfa y beta , Transducción de Señal , Trofoblastos , Infección por el Virus Zika , Virus Zika , Femenino , Animales , Embarazo , Interferón Tipo I/metabolismo , Interferón Tipo I/inmunología , Transducción de Señal/inmunología , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Placenta/inmunología , Placenta/virología , Placenta/metabolismo , Infección por el Virus Zika/inmunología , Infección por el Virus Zika/virología , Virus Zika/inmunología , Virus Zika/fisiología , Ratones , Decidua/inmunología , Decidua/virología , Decidua/metabolismo , Feto/inmunología , Feto/virología , Trofoblastos/inmunología , Trofoblastos/virología , Trofoblastos/metabolismo , Receptor de Interferón alfa y beta/genética , Receptor de Interferón alfa y beta/metabolismo , Ratones Endogámicos C57BL , Ratones Noqueados , Inmunidad Innata , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/virología , Modelos Animales de EnfermedadRESUMEN
Vertical transmission of the Zika virus (ZIKV) causes severe fetal defects, but the exact pathogenic mechanism is unclear. We identified up to a 10,480-fold higher expression of viral attachment factors AXL, GAS6, and PROS1 and a 3880-fold increase in ZIKV infectiousness/propagation in human term decidual stromal cells versus trophoblasts. Moreover, levels of viral attachment factors and ZIKV are significantly increased, whereas expression of innate immune response genes are significantly decreased, in human first trimester versus term decidual cells. ZIKV-infected decidual cell supernatants increased cytotrophoblasts infection up to 252-fold compared with directly infected cytotrophoblasts. Tizoxanide treatment efficiently inhibited Zika infection in both maternal and fetal cells. We conclude that ZIKV permissiveness, as well as innate immune responsiveness of human decidual cells, are gestational age dependent, and decidual cells augment ZIKV infection of primary human cytotrophoblast cultures, which are otherwise ZIKV resistant. Human decidual cells may act as reservoirs for trimester-dependent placental transmission of ZIKV, accounting for the higher Zika infection susceptibility and more severe fetal sequelae observed in early versus late pregnancy. Moreover, tizoxanide is a promising agent in preventing perinatal Zika transmission as well as other RNA viruses such as coronavirus.
Asunto(s)
Decidua , Edad Gestacional , Inmunidad Innata , Transmisión Vertical de Enfermedad Infecciosa , Complicaciones Infecciosas del Embarazo , Infección por el Virus Zika , Virus Zika/inmunología , Animales , Chlorocebus aethiops , Decidua/inmunología , Decidua/patología , Decidua/virología , Femenino , Humanos , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/patología , Trofoblastos , Células Vero , Infección por el Virus Zika/inmunología , Infección por el Virus Zika/patología , Infección por el Virus Zika/transmisiónRESUMEN
The current Zika virus (ZIKV) outbreak is associated with neurological malformations and disorders in neonates. Areas of increased incidence of malformations may overlap with dengue-hyperendemic areas. ZIKV infection is enhanced by antibodies against dengue virus (DENV) in cell culture and inbred mice. Sufficiently powered clinical studies or primate studies addressing the enhancement of fetal ZIKV infection after previous dengue infection are not available. The human placenta is susceptible to ZIKV in vitro, but it is unknown whether antibody-dependent enhancement of ZIKV infection occurs at the placental barrier. Here we studied ZIKV infection in placental tissue in the presence of DENV-immune sera. Explants from the amniochorionic membrane, the chorionic villi, and the maternal decidua were infected with ZIKV in the presence of DENV type 1-, 2-, or 4-immune sera, or controls. Presence of DENV antibodies of any type enhanced the percentage of successful infections of organ explants between 1.42- and 2.67-fold, and led to a faster replication as well as significantly increased virus production. No enhancement was seen with yellow fever or chikungunya virus control sera. Pre-existing DENV antibodies may pose an increased risk of trans-placental ZIKV transmission.
Asunto(s)
Anticuerpos Antivirales/inmunología , Placenta/citología , Placenta/virología , Infección por el Virus Zika/inmunología , Amnios/citología , Amnios/virología , Acrecentamiento Dependiente de Anticuerpo , Vellosidades Coriónicas/virología , Decidua/citología , Decidua/virología , Virus del Dengue , Femenino , Humanos , Técnicas de Cultivo de Órganos , Placenta/inmunología , Embarazo , ARN Viral , Técnicas de Cultivo de Tejidos , Virus Zika/patogenicidadRESUMEN
Hepatitis E virus (HEV) infection, particularly HEV genotype 1 (HEV-1), can result in fulminant hepatic failure and severe placental diseases, but mechanisms underlying genotype-specific pathogenicity are unclear and appropriate models are lacking. Here, we model HEV-1 infection ex vivo at the maternal-fetal interface using the decidua basalis and fetal placenta, and compare its effects to the less-pathogenic genotype 3 (HEV-3). We demonstrate that HEV-1 replicates more efficiently than HEV-3 both in tissue explants and stromal cells, produces more infectious progeny virions and causes severe tissue alterations. HEV-1 infection dysregulates the secretion of several soluble factors. These alterations to the cytokine microenvironment correlate with viral load and contribute to the tissue damage. Collectively, this study characterizes an ex vivo model for HEV infection and provides insights into HEV-1 pathogenesis during pregnancy that are linked to high viral replication, alteration of the local secretome and induction of tissue injuries.
Asunto(s)
Virus de la Hepatitis E/genética , Virus de la Hepatitis E/patogenicidad , Intercambio Materno-Fetal/fisiología , Células Cultivadas , Decidua/patología , Decidua/virología , Femenino , Genotipo , Humanos , Interferones/metabolismo , Embarazo , Células del Estroma/metabolismo , Replicación Viral , Interferón lambdaRESUMEN
OBJECTIVES: Mesenchymal stromal/stem cells have immunosuppressive functions. Our previous results demonstrated that one of the players of this immunomodulation can be ascribed to the Human Leukocyte Antigen-G. HLA-G, a non classical HLA class I antigen, is involved in immune tolerance during pregnancy, organ transplantation, autoimmune and inflammatory diseases. In this study we wanted to verify whether human endometrial decidual tissue derived (EDT)-MSC could express HLA-G. Additionally we assessed the permissivity to Human Herpesvirus infections, using HSV-1 as a model, and the possible effect on EDT-MSC immunosuppressive functions towards peripheral blood mononuclear cell (PBMC) proliferation. METHODS: We analyzed immune-inhibitory functions and HLA-G expression in human EDT-MSC before and after HSV-1 infection. RESULTS: We observed that EDT-MSC express HLA-G molecules, that partly are responsible for the immune-inhibitory functions of EDT-MSC towards PBMC proliferation. EDT-MSC are permissive for a productive infection by HSV-1, that decreases HLA-G expression and affects EDT-MSC immune-inhibitory functions. CONCLUSIONS: We demonstrate that EDT-MSC are susceptible to HSV-1 infection, that reduces HLA-G expression and their immune-inhibitory function. These data could have a clinical implication in the use of EDT-MSC as an immunosuppressant, in particular in steroid-refractory GvHD after allogeneic hematopoietic stem cell transplantation and in autoimmune diseases.
Asunto(s)
Decidua/citología , Decidua/virología , Antígenos HLA-G/genética , Herpes Simple/genética , Herpes Simple/virología , Herpesvirus Humano 1 , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/virología , Biomarcadores , Supervivencia Celular , Decidua/inmunología , Femenino , Regulación de la Expresión Génica , Antígenos HLA-G/inmunología , Herpes Simple/inmunología , Herpesvirus Humano 1/inmunología , Humanos , Inmunomodulación , InmunofenotipificaciónRESUMEN
Human cytomegalovirus (HCMV) is the leading cause of congenital infection and is associated with a wide range of neurodevelopmental disabilities and intrauterine growth restriction. Yet our current understanding of the mechanisms modulating transplacental HCMV transmission is poor. The placenta, given its critical function in protecting the fetus, has evolved effective yet largely uncharacterized innate immune barriers against invading pathogens. Here we show that the intrinsic cellular restriction factor apolipoprotein B editing catalytic subunit-like 3A (APOBEC3A [A3A]) is profoundly upregulated following ex vivo HCMV infection in human decidual tissues-constituting the maternal aspect of the placenta. We directly demonstrated that A3A severely restricted HCMV replication upon controlled overexpression in epithelial cells, acting by a cytidine deamination mechanism to introduce hypermutations into the viral genome. Importantly, we further found that A3 editing of HCMV DNA occurs both ex vivo in HCMV-infected decidual organ cultures and in vivo in amniotic fluid samples obtained during natural congenital infection. Our results reveal a previously unexplored role for A3A as an innate anti-HCMV effector, activated by HCMV infection in the maternal-fetal interface. These findings pave the way to new insights into the potential impact of APOBEC proteins on HCMV pathogenesis.IMPORTANCE In view of the grave outcomes associated with congenital HCMV infection, there is an urgent need to better understand the innate mechanisms acting to limit transplacental viral transmission. Toward this goal, our findings reveal the role of the intrinsic cellular restriction factor A3A (which has never before been studied in the context of HCMV infection and vertical viral transmission) as a potent anti-HCMV innate barrier, activated by HCMV infection in the authentic tissues of the maternal-fetal interface. The detection of naturally occurring hypermutations in clinical amniotic fluid samples of congenitally infected fetuses further supports the idea of the occurrence of A3 editing of the viral genome in the setting of congenital HCMV infection. Given the widely differential tissue distribution characteristics and biological functions of the members of the A3 protein family, our findings should pave the way to future studies examining the potential impact of A3A as well as of other A3s on HCMV pathogenesis.
Asunto(s)
Citidina Desaminasa/genética , Citidina Desaminasa/metabolismo , Infecciones por Citomegalovirus/virología , Decidua/inmunología , Inmunidad Innata , Placenta/inmunología , Proteínas/genética , Proteínas/metabolismo , Líquido Amniótico/inmunología , Líquido Amniótico/virología , Citidina Desaminasa/inmunología , Citomegalovirus/genética , Citomegalovirus/inmunología , Citomegalovirus/patogenicidad , Infecciones por Citomegalovirus/congénito , Infecciones por Citomegalovirus/inmunología , Decidua/citología , Decidua/virología , Femenino , Edición Génica , Genoma Viral , Humanos , Recién Nacido , Transmisión Vertical de Enfermedad Infecciosa , Técnicas de Cultivo de Órganos , Placenta/citología , Placenta/virología , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/virología , Proteínas/inmunología , Regulación hacia Arriba , Replicación ViralRESUMEN
Infection with Zika virus (ZIKV) is associated with human congenital fetal anomalies. To model fetal outcomes in nonhuman primates, we administered Asian-lineage ZIKV subcutaneously to four pregnant rhesus macaques. While non-pregnant animals in a previous study contemporary with the current report clear viremia within 10-12 days, maternal viremia was prolonged in 3 of 4 pregnancies. Fetal head growth velocity in the last month of gestation determined by ultrasound assessment of head circumference was decreased in comparison with biparietal diameter and femur length within each fetus, both within normal range. ZIKV RNA was detected in tissues from all four fetuses at term cesarean section. In all pregnancies, neutrophilic infiltration was present at the maternal-fetal interface (decidua, placenta, fetal membranes), in various fetal tissues, and in fetal retina, choroid, and optic nerve (first trimester infection only). Consistent vertical transmission in this primate model may provide a platform to assess risk factors and test therapeutic interventions for interruption of fetal infection. The results may also suggest that maternal-fetal ZIKV transmission in human pregnancy may be more frequent than currently appreciated.
Asunto(s)
Transmisión Vertical de Enfermedad Infecciosa , Complicaciones Infecciosas del Embarazo , Infección por el Virus Zika/transmisión , Virus Zika/fisiología , Líquido Amniótico/virología , Animales , Decidua/patología , Decidua/virología , Modelos Animales de Enfermedad , Femenino , Desarrollo Fetal , Feto , Humanos , Pulmón/patología , Pulmón/virología , Macaca mulatta , Placenta/patología , Placenta/virología , Embarazo , ARN Viral/análisis , Bazo/patología , Bazo/virología , Cordón Umbilical/patología , Cordón Umbilical/virología , Viremia , Infección por el Virus Zika/patología , Infección por el Virus Zika/virologíaRESUMEN
Zika virus (ZIKV) has emerged as a cause of congenital brain anomalies and a range of placenta-related abnormalities, highlighting the need to unveil the modes of maternal-fetal transmission. The most likely route of vertical ZIKV transmission is via the placenta. The earliest events of ZIKV transmission in the maternal decidua, representing the maternal uterine aspect of the chimeric placenta, have remained unexplored. Here, we show that ZIKV replicates in first-trimester human maternal-decidual tissues grown ex vivo as three-dimensional (3D) organ cultures. An efficient viral spread in the decidual tissues was demonstrated by the rapid upsurge and continued increase of tissue-associated ZIKV load and titers of infectious cell-free virus progeny, released from the infected tissues. Notably, maternal decidual tissues obtained at midgestation remained similarly susceptible to ZIKV, whereas fetus-derived chorionic villi demonstrated reduced ZIKV replication with increasing gestational age. A genome-wide transcriptome analysis revealed that ZIKV substantially upregulated the decidual tissue innate immune responses. Further comparison of the innate tissue response patterns following parallel infections with ZIKV and human cytomegalovirus (HCMV) revealed that unlike HCMV, ZIKV did not induce immune cell activation or trafficking responses in the maternal-fetal interface but rather upregulated placental apoptosis and cell death molecular functions. The data identify the maternal uterine aspect of the human placenta as a likely site of ZIKV transmission to the fetus and further reveal distinct patterns of innate tissue responses to ZIKV. Our unique experimental model and findings could further serve to study the initial stages of congenital ZIKV transmission and pathogenesis and evaluate the effect of new therapeutic interventions. IMPORTANCE: In view of the rapid spread of the current ZIKV epidemic and the severe manifestations of congenital ZIKV infection, it is crucial to learn the fundamental mechanisms of viral transmission from the mother to the fetus. Our studies of ZIKV infection in the authentic tissues of the human maternal-fetal interface unveil a route of transmission whereby virus originating from the mother could reach the fetal compartment via efficient replication within the maternal decidual aspect of the placenta, coinhabited by maternal and fetal cells. The identified distinct placental tissue innate immune responses and damage pathways could provide a mechanistic basis for some of the placental developmental abnormalities associated with ZIKV infection. The findings in the unique model of the human decidua should pave the way to future studies examining the interaction of ZIKV with decidual immune cells and to evaluation of therapeutic interventions aimed at the earliest stages of transmission.
Asunto(s)
Decidua/virología , Inmunidad Innata , Placenta/virología , Complicaciones Infecciosas del Embarazo , Infección por el Virus Zika/inmunología , Infección por el Virus Zika/virología , Virus Zika/fisiología , Animales , Línea Celular , Vellosidades Coriónicas/virología , Citomegalovirus/inmunología , Infecciones por Citomegalovirus/inmunología , Infecciones por Citomegalovirus/transmisión , Infecciones por Citomegalovirus/virología , Susceptibilidad a Enfermedades , Femenino , Expresión Génica , Edad Gestacional , Humanos , Transmisión Vertical de Enfermedad Infecciosa , Interferones/genética , Interferones/metabolismo , Embarazo , Transducción de Señal , Infección por el Virus Zika/metabolismo , Infección por el Virus Zika/transmisiónRESUMEN
The combination of the activating killer cell Ig-like receptor 2DS1 (KIR2DS1) expressed by maternal decidual natural killer cells (dNK) and the presence of its ligand, the HLA-C allotype HLA-C2, expressed by fetal trophoblasts, reduces the risk of developing pregnancy complications. However, no molecular or cellular mechanism explains this genetic correlation. Here we demonstrate that KIR2DS1+ dNK acquired higher cytotoxic function than KIR2DS1- dNK when exposed to human cytomegalovirus (HCMV)-infected decidual stromal cells (DSC), particularly when DSCs express HLA-C2. Furthermore, dNK were unable to degranulate or secrete cytokines in response to HCMV-infected primary fetal extravillous trophoblasts. This emphasizes the immunological challenge to clear placental viral infections within the immune-privileged placenta. Activation of dNK through KIR2DS1/HLA-C2 interaction increases their ability to respond to placental HCMV infection and may limit subsequent virus-induced placental pathology. This mechanism is directly related to how KIR2DS1 expressed by dNK reduces development of severe pregnancy complications such as miscarriages and preterm delivery.
Asunto(s)
Infecciones por Citomegalovirus/inmunología , Decidua/inmunología , Células Asesinas Naturales/inmunología , Placenta/inmunología , Receptores KIR/metabolismo , Separación Celular , Citomegalovirus , Decidua/virología , Femenino , Citometría de Flujo , Perfilación de la Expresión Génica , Antígenos HLA-C/análisis , Humanos , Tolerancia Inmunológica , Inmunoglobulinas/metabolismo , Placenta/virología , Placentación , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Complicaciones Infecciosas del Embarazo/virología , Resultado del Embarazo , Receptores KIR/genética , Trofoblastos/virologíaRESUMEN
The initial interplay between human cytomegalovirus (HCMV) and innate tissue response in the human maternal-fetal interface, though crucial for determining the outcome of congenital HCMV infection, has remained unknown. We studied the innate response to HCMV within the milieu of the human decidua, the maternal aspect of the maternal-fetal interface, maintained ex vivo as an integral tissue. HCMV infection triggered a rapid and robust decidual-tissue innate immune response predominated by interferon (IFN)γ and IP-10 induction, dysregulating the decidual cytokine/chemokine environment in a distinctive fashion. The decidual-tissue response was already elicited during viral-tissue contact, and was not affected by neutralizing HCMV antibodies. Of note, IFNγ induction, reflecting immune-cell activation, was distinctive to the maternal decidua, and was not observed in concomitantly-infected placental (fetal) villi. Our studies in a clinically-relevant surrogate human model, provide a novel insight into the first-line decidual tissue response which could affect the outcome of congenital infection.
Asunto(s)
Infecciones por Citomegalovirus/inmunología , Citomegalovirus/inmunología , Interacciones Huésped-Patógeno/inmunología , Inmunidad Innata , Placenta/inmunología , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Citocinas/genética , Citocinas/metabolismo , Infecciones por Citomegalovirus/genética , Infecciones por Citomegalovirus/metabolismo , Infecciones por Citomegalovirus/virología , Decidua/inmunología , Decidua/metabolismo , Decidua/patología , Decidua/virología , Femenino , Expresión Génica , Heparina/metabolismo , Humanos , Mediadores de Inflamación/metabolismo , Placenta/metabolismo , Placenta/patología , Placenta/virología , Embarazo , Virión/inmunologíaRESUMEN
Toll-like receptors (TLRs) form the major family of pattern recognition receptors (PRRs) that are involved in innate immunity. Innate immune responses against microorganisms at the maternal-fetal interface may have a significant impact on the success of pregnancy, as intrauterine infections have been shown to be strongly associated with certain complications of pregnancy. At the maternal-fetal interface, TLRs are expressed not only in the immune cells but also in non-immune cells such as trophoblasts and decidual cells; moreover, their expression patterns vary according to the stage of pregnancy. Here, we will update potential functions of TLRs in these cells, their recognition and response to microorganisms, and their involvement in the innate immunity. The impact of TLR-mediated innate immune response will be discussed via animal model studies, as well as clinical observations.
Asunto(s)
Pérdida del Embrión/inmunología , Intercambio Materno-Fetal/inmunología , Trabajo de Parto Prematuro/inmunología , Preeclampsia/inmunología , Complicaciones Infecciosas del Embarazo/inmunología , Embarazo/inmunología , Receptores Toll-Like/inmunología , Animales , Infecciones Bacterianas/inmunología , Infecciones Bacterianas/microbiología , Infecciones Bacterianas/patología , Infecciones Bacterianas/virología , Decidua/inmunología , Decidua/microbiología , Decidua/patología , Decidua/virología , Pérdida del Embrión/microbiología , Pérdida del Embrión/patología , Pérdida del Embrión/virología , Femenino , Regulación de la Expresión Génica , Humanos , Trabajo de Parto Prematuro/microbiología , Trabajo de Parto Prematuro/patología , Trabajo de Parto Prematuro/virología , Preeclampsia/microbiología , Preeclampsia/patología , Preeclampsia/virología , Complicaciones Infecciosas del Embarazo/microbiología , Complicaciones Infecciosas del Embarazo/patología , Complicaciones Infecciosas del Embarazo/virología , Transducción de Señal , Receptores Toll-Like/genética , Trofoblastos/inmunología , Trofoblastos/microbiología , Trofoblastos/patología , Trofoblastos/virología , Virosis/inmunología , Virosis/microbiología , Virosis/patología , Virosis/virologíaRESUMEN
During the first trimester of pregnancy the uterus is massively infiltrated by decidual natural killer cells (dNK). These cells are not killers, but they rather provide a microenvironment that is propitious to healthy placentation. Human cytomegalovirus (HCMV) is the most common cause of intrauterine viral infections and a known cause of severe birth defects or fetal death. The rate of HCMV congenital infection is often low in the first trimester of pregnancy. The mechanisms controlling HCMV spreading during pregnancy are not yet fully revealed, but evidence indicating that the innate immune system plays a role in controlling HCMV infection in healthy adults exists. In this study, we investigated whether dNK cells could be involved in controlling viral spreading and in protecting the fetus against congenital HCMV infection. We found that freshly isolated dNK cells acquire major functional and phenotypic changes when they are exposed to HCMV-infected decidual autologous fibroblasts. Functional studies revealed that dNK cells, which are mainly cytokines and chemokines producers during normal pregnancy, become cytotoxic effectors upon their exposure to HCMV-infected autologous decidual fibroblasts. Both the NKG2D and the CD94/NKG2C or 2E activating receptors are involved in the acquired cytotoxic function. Moreover, we demonstrate that CD56(pos) dNK cells are able to infiltrate HCMV-infected trophoblast organ culture ex-vivo and to co-localize with infected cells in situ in HCMV-infected placenta. Taken together, our results present the first evidence suggesting the involvement of dNK cells in controlling HCMV intrauterine infection and provide insights into the mechanisms through which these cells may operate to limit the spreading of viral infection to fetal tissues.
Asunto(s)
Infecciones por Citomegalovirus/inmunología , Citomegalovirus/inmunología , Decidua/inmunología , Células Asesinas Naturales/inmunología , Antígeno CD56/metabolismo , Línea Celular , Infecciones por Citomegalovirus/congénito , Infecciones por Citomegalovirus/metabolismo , Decidua/citología , Decidua/virología , Proteína Ligando Fas/metabolismo , Femenino , Células HEK293 , Humanos , Células Asesinas Naturales/metabolismo , Subfamília C de Receptores Similares a Lectina de Células NK/inmunología , Subfamília C de Receptores Similares a Lectina de Células NK/metabolismo , Subfamília D de Receptores Similares a Lectina de las Células NK/inmunología , Subfamília D de Receptores Similares a Lectina de las Células NK/metabolismo , Subfamilia K de Receptores Similares a Lectina de Células NK/inmunología , Subfamilia K de Receptores Similares a Lectina de Células NK/metabolismo , Placentación , Embarazo , Primer Trimestre del Embarazo , Ligando Inductor de Apoptosis Relacionado con TNF/metabolismo , ÚteroRESUMEN
Human cytomegalovirus (HCMV) is the leading cause of congenital infection, associated with severe birth defects and intrauterine growth retardation. The mechanism of HCMV transmission via the maternal-fetal interface is largely unknown, and there are no animal models for HCMV. The initial stages of infection are believed to occur in the maternal decidua. Here we employed a novel decidual organ culture, using both clinically derived and laboratory-derived viral strains, for the ex vivo modeling of HCMV transmission in the maternal-fetal interface. Viral spread in the tissue was demonstrated by the progression of infected-cell foci, with a 1.3- to 2-log increase in HCMV DNA and RNA levels between days 2 and 9 postinfection, the expression of immediate-early and late proteins, the appearance of typical histopathological features of natural infection, and dose-dependent inhibition of infection by ganciclovir and acyclovir. HCMV infected a wide range of cells in the decidua, including invasive cytotrophoblasts, macrophages, and endothelial, decidual, and dendritic cells. Cell-to-cell viral spread was revealed by focal extension of infected-cell clusters, inability to recover infectious extracellular virus, and high relative proportions (88 to 93%) of cell-associated viral DNA. Intriguingly, neutralizing HCMV hyperimmune globulins exhibited inhibitory activity against viral spread in the decidua even when added at 24 h postinfection-providing a mechanistic basis for their clinical use in prenatal prevention. The ex vivo-infected decidual cultures offer unique insight into patterns of viral tropism and spread, defining initial stages of congenital HCMV transmission, and can facilitate evaluation of the effects of new antiviral interventions within the maternal-fetal interface milieu.
Asunto(s)
Infecciones por Citomegalovirus/transmisión , Decidua/virología , Transmisión Vertical de Enfermedad Infecciosa , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , ADN Viral/genética , ADN Viral/aislamiento & purificación , Femenino , Expresión Génica , Humanos , Modelos Biológicos , Técnicas de Cultivo de Órganos/métodos , Embarazo , ARN Viral/genética , ARN Viral/aislamiento & purificación , Factores de Tiempo , Proteínas Virales/biosíntesisRESUMEN
BACKGROUND: Maternofetal transmission (MFT) of HIV-1 is relatively rare during the first trimester of pregnancy despite the permissivity of placental cells for cell-to-cell HIV-1 infection. Invasive placental cells interact directly with decidual cells of the uterine mucosa during the first months of pregnancy, but the role of the decidua in the control of HIV-1 transmission is unknown. RESULTS: We found that decidual mononuclear cells naturally produce low levels of IL-10, IL-12, IL-15, TNF-α, IFN-α, IFN-γ and CXCL-12 (SDF-1), and large amounts of CCL-2 (MCP1), CCL-3 (MIP-1α), CCL-4 (MIP-1ß), CCL-5 (Rantes), CXCL-10 (IP-10), IL-6 and IL-8. CCL-3 and CCL-4 levels were significantly upregulated by in vitro infection with R5 HIV-1 but not X4. Decidual CD14+ antigen presenting cells were the main CCL-3 and CCL-4 producers among decidual leukocytes. R5 and X4 HIV-1 infection was inhibited by decidual cell culture supernatants in vitro. Using HIV-1 pseudotypes, we found that inhibition of the HIV-1 entry step was inhibited by decidual soluble factors. CONCLUSION: Our findings show that decidual innate immunity (soluble factors) is involved in the control of HIV-1 infection at the maternofetal interface. The decidua could thus serve as a mucosal model for identifying correlates of protection against HIV-1 infection.
Asunto(s)
Citocinas/inmunología , Decidua/inmunología , Infecciones por VIH/transmisión , VIH-1/fisiología , Transmisión Vertical de Enfermedad Infecciosa , Complicaciones Infecciosas del Embarazo/inmunología , Citocinas/genética , Decidua/virología , Femenino , Infecciones por VIH/genética , Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-1/inmunología , Humanos , Embarazo , Complicaciones Infecciosas del Embarazo/virologíaRESUMEN
NK cells present in the peripheral blood respond rapidly to pathogens or pathogen-infected cells by various means including cytotoxicity and production of cytokines. Whether decidual NK (dNK) cells are able to play a similar role when the pregnant uterus is infected by viruses is still largely unknown. Decidual NK cells are generally considered as poorly cytotoxic when compared to their peripheral blood counterparts. However, we have recently demonstrated that freshly isolated dNK cells from healthy early pregnant uterus do have a cytotoxic potential mediated by the specific engagement of NKp46 activating receptor. We further found that the co-engagement of CD94/NKG2A inhibiting receptor drastically inhibits the cytolytic function of dNK. This latter observation suggests that in situ the CD94/NKG2A receptor interaction with its HLA-E specific ligand is a dominant negative regulatory mechanism that prevents unwanted dNK cell cytotoxicity in non-infected pregnant uterus. How do dNK cells behave when they are activated by virus-infected cells present at the maternal-fetal interface? Largely based on data obtained from circulating NK cells, this review briefly discusses the following questions: Does uterine viral infection promote decidual NK cell proliferative capacity in situ? Are dNK cells able to kill virus-infected autologous decidual target cells and thus limit the virus spreading to the fetus? Which viral-mediated signal(s) and molecular interactions may subvert inhibition of dNK cytotoxic potential? Does uterine viral infection promote decidual NK cell secretion of cytokines and chemokines that boost the anti-viral immune response?
Asunto(s)
Citotoxicidad Inmunológica , Decidua/inmunología , Células Asesinas Naturales/inmunología , Complicaciones Infecciosas del Embarazo/inmunología , Virosis/inmunología , Animales , Presentación de Antígeno , Antígenos Virales/inmunología , Circulación Sanguínea , Decidua/virología , Femenino , Humanos , Evasión Inmune , Activación de Linfocitos , Circulación Placentaria , EmbarazoRESUMEN
Pregnancy failure is a common event and often of unknown cause. Some viruses are thought to cause abortions including the adeno-associated viruses (AAV), viruses which are regarded as being without any definitive association to any human disease. This study investigated AAV infection in 81 human abortions, both spontaneous and intentional that occurred up to the 23rd week of gestation. Nucleic acid of AAV-2, 3, and 5 types from 118 decidual and chorionic tissues, collected from the patients in this study, was amplified by nested-PCR. In situ hybridization (ISH) was developed with a digoxigenin-labeled AAV probe in paraffin embedded tissues from the AAV positive cases. AAV was observed in 28.4% (23/81) of the cases, of which, 78.3% (18/23) were in the decidua and 21.7% (5/23) in the extravillous trophoblast, the chorionic plate, or chorionic villi fragments. AAV-2, the only type detected, occurred in 32.3% (22/68) and in 7.7% (1/13) of the spontaneous and intentional abortions, respectively. ISH revealed AAV in the decidua, chorionic tissue or chorionic plate and extravillous trophoblast. The detection of only AAV-2 type indicates that it is the most frequent in the population studied and/or shows tissue tropism. The presence of AAV in decidual or trophoblastic cells in cases of abortion, as observed by ISH, implies that the virus could jeopardize the pregnancy. The significant predominance in spontaneous cases suggests possibly a causal association between AAV and abortion.
Asunto(s)
Aborto Espontáneo/etiología , Dependovirus/aislamiento & purificación , Infecciones por Parvoviridae/complicaciones , Infecciones por Parvoviridae/diagnóstico , Aborto Espontáneo/virología , Corion/virología , ADN Viral/genética , ADN Viral/aislamiento & purificación , Decidua/virología , Femenino , Humanos , Hibridación in Situ/métodos , Recién Nacido , Infecciones por Parvoviridae/virología , Patología Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Embarazo , Trofoblastos/virología , Virología/métodosRESUMEN
Human fetal membranes are composed of amnion, chorion and decidua tissues, which play a critical role in defense barriers as well as maintenance of pregnancy and parturition. Pro-inflammatory cytokines, such as interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha, produced by the tissues are postulated to facilitate parturition. Influenza virus infection is one of causes of pregnancy-associated complications, such as premature delivery, abortion and stillbirth. Recent studies have demonstrated that influenza virus infection induced the gene expression of a set of pro-inflammatory cytokines, such as IL-1beta, IL-6, TNF-alpha, interferon (IFN)-beta, IFN-gamma and granulocyte macrophage colony-stimulating factor (GM-CSF), and the secretion of unidentified monocyte differentiation-inducing factor(s) from primary cultured chorion cells undergoing apoptosis. These phenomena were not observed in primary cultured amnion cells infected with the virus. This article reviews, (1) the production of cytokines in fetal membrane tissues and their functions; (2) the differential induction of pro-inflammatory cytokine gene expression and apoptosis in fetal membrane chorion and amnion cells by influenza virus infection. An accumulating number of evidence suggests that interactive reactions between fetal membrane chorion cells and maternal monocytes/macrophages may play a critical role in defense barriers against the virus infection. Understanding the interactions would make important contributions to the elucidation of the pathogenesis of influenza virus infection during pregnancy.
Asunto(s)
Apoptosis , Citocinas/biosíntesis , Membranas Extraembrionarias/inmunología , Gripe Humana/complicaciones , Complicaciones Infecciosas del Embarazo/inmunología , Amnios/inmunología , Amnios/virología , Corion/inmunología , Corion/virología , Citocinas/genética , Decidua/inmunología , Decidua/virología , Membranas Extraembrionarias/virología , Femenino , Humanos , Gripe Humana/inmunología , Embarazo , Complicaciones Infecciosas del Embarazo/metabolismoRESUMEN
AIM: To determine the expression of CMV-associated antigen in the human decidual endometrial stromal cells in spontaneous abortions with no evidence of maternal relapse during the first trimester of gestation. EXPERIMENTAL DESIGN: We examined 15 placentas resulting from intrauterine fetal death after spontaneous abortion during the 8th, 10th, and 12th week of gestation respectively, and in which CMV reactivation was ruled out from serological evaluation of the pregnant women at admission, versus equal controls after voluntary abortion following well-documented maternal viral recurrence. In addition, a panel of monoclonal antibodies for the identification of leukocytes (CD45/LCA), B-lymphocytes (CD20/L-26), and T-lymphocytes (CD45RO/UCHL1), was performed. All women received hormonal medication to support gestation, in the cases of spontaneous abortions. RESULTS: Immunohistochemical examination using a specific antibody against cytomegalovirus showed large multinucleated infected cells with intranuclear inclusions, located primarily in the decidual stroma within a lymphoplasmacytic infiltrate in the cases of spontaneous abortions. No evidence of infection was observed in the chorionic villi. In the cases of voluntary abortions same findings were observed in the relevant areas, and a strong evidence of infection was observed in the chorionic villi. CONCLUSION: This study demonstrates 1) that the decidual endometrial stromal cells can express the CMV-associated antigen prior to serological manifestation of the viral replication, 2) the expression of the antigen is higher in cases of hormonal administration to support gestation. In these cases a mild mononuclear infiltrate of UCHL1 (T marker) positive cells, accompanies the CMV-associated antigen positive cells.
Asunto(s)
Aborto Espontáneo/virología , Antígenos Virales/análisis , Citomegalovirus/aislamiento & purificación , Decidua/virología , Aborto Espontáneo/patología , Infecciones por Citomegalovirus/complicaciones , Infecciones por Citomegalovirus/diagnóstico , Femenino , Humanos , Inmunohistoquímica , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Primer Trimestre del EmbarazoRESUMEN
Prenatal cytomegalovirus infection may cause pregnancy complications such as intrauterine growth restriction and birth defects. How virus from the mother traverses the placenta is unknown. PCR analysis of biopsy specimens of the maternal-fetal interface revealed that DNA sequences from cytomegalovirus were commonly found with those of herpes simplex viruses and pathogenic bacteria. Cytomegalovirus DNA and infected cell proteins were found more often in the decidua than in the placenta, suggesting that the uterus functions as a reservoir for infection. In women with low neutralizing titers, cytomegalovirus replicated in diverse decidual cells and placental trophoblasts and capillaries. In women with intermediate to high neutralizing titers, decidual infection was suppressed and the placenta was spared. Overall, cytomegalovirus virions and maternal immunoglobulin G were detected in syncytiotrophoblasts, villus core macrophages, and dendritic cells. These results suggest that the outcome of cytomegalovirus infection depends on the presence of other pathogens and coordinated immune responses to viral replication at the maternal-fetal interface.
Asunto(s)
Bacterias/aislamiento & purificación , Infecciones por Citomegalovirus/inmunología , Infecciones por Citomegalovirus/transmisión , Transmisión Vertical de Enfermedad Infecciosa , Placenta/virología , Útero/virología , Bacterias/genética , Bacterias/patogenicidad , Infecciones Bacterianas/microbiología , Citomegalovirus/genética , Citomegalovirus/aislamiento & purificación , Infecciones por Citomegalovirus/virología , Decidua/microbiología , Decidua/virología , Femenino , Humanos , Placenta/microbiología , Enfermedades Placentarias/virología , Embarazo , Complicaciones Infecciosas del Embarazo/virología , Primer Trimestre del Embarazo , Segundo Trimestre del Embarazo , Útero/microbiologíaRESUMEN
OBJECTIVE: To investigate the pathogenic factors of human cytomegalovirus (HCMV) infections. METHODS: Totally 36 serum samples were obtained from early pregnant woman and examined with ELISA for anti-HCMV antibody IgG and IgM. After artificial abortion,chorionic villus and decidua were also examined with polymerase chain reaction (PCR) for HCMV-DNA. When the results of PCR were positive, pathological changes of these chorionic villus and decidua were analyzed. RESULTS: The results showed that only 10 samples were PCR positive while IgG and/or IgM antibody to HCMV was positive. After infection with HCMV, different changes occurred in chorionic villus and decidual trophoblastic cells placental villus were hyperplasic and decidua cells degenerated and necrotized followed by lymphocytes infiltration. CONCLUSION: These pathological changes may be one of pathogenic factors of HCMV.