scholarly journals Human cytomegalovirus infection changes the pattern of surface markers of small extracellular vesicles isolated from first trimester placental histocultures

2020 ◽  
Author(s):  
Mathilde Bergamelli ◽  
Hélène Martin ◽  
Mélinda Bénard ◽  
Jérôme Ausseil ◽  
Jean-Michel Mansuy ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Human Cytomegalovirus ◽  
Predictive Biomarkers ◽  
Congenital Infection ◽  
First Trimester ◽  
Maternal Serum ◽  
Surface Markers ◽  
Non Invasive ◽  
Diagnosis And Prognosis ◽  
Human Cytomegalovirus Infection

ABSTRACTCurrently, research on the use of non-invasive biomarkers as diagnosis and prognosis tools during pathological pregnancies is in full development. Among these, placenta-derived small extracellular vesicles (sEVs) are considered as serious candidates, since their composition is modified during many pregnancy pathologies. Moreover, sEVs are found in maternal serum and can thus be easily purified from a simple blood sample. In this study, we describe the isolation of sEVs from a histoculture model of first trimester placental explants. Using bead-based multiplex cytometry and electron microscopy combined with biochemical approaches, we characterized these sEVs and defined their associated markers and ultrastructure. We next examined the consequences of infection by human cytomegalovirus on sEVs secretion and characteristics. We observed that infection led to increased levels of expression of several surface markers, without any impact on the secretion and integrity of sEVs. Our findings open the prospect for the identification of new predictive biomarkers for the severity and outcome of this congenital infection early during pregnancy, which are still sorely lacking.

scholarly journals Human Cytomegalovirus Infection Changes the Pattern of Surface Markers of Small Extracellular Vesicles Isolated From First Trimester Placental Long-Term Histocultures

2021 ◽  
Vol 9 ◽  
Author(s):  
Mathilde Bergamelli ◽  
Hélène Martin ◽  
Mélinda Bénard ◽  
Jérôme Ausseil ◽  
Jean-Michel Mansuy ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Human Cytomegalovirus ◽  
Predictive Biomarkers ◽  
Congenital Infection ◽  
First Trimester ◽  
Surface Markers ◽  
Human Cytomegalovirus Infection ◽  
Adverse Pregnancy

Extracellular vesicles (EVs) have increasingly been recognized as key players in a wide variety of physiological and pathological contexts, including during pregnancy. Notably, EVs appear both as possible biomarkers and as mediators involved in the communication of the placenta with the maternal and fetal sides. A better understanding of the physiological and pathological roles of EVs strongly depends on the development of adequate and reliable study models, specifically at the beginning of pregnancy where many adverse pregnancy outcomes have their origin. In this study, we describe the isolation of small EVs from a histoculture model of first trimester placental explants in normal conditions as well as upon infection by human cytomegalovirus. Using bead-based multiplex cytometry and electron microscopy combined with biochemical approaches, we characterized these small EVs and defined their associated markers and ultrastructure. We observed that infection led to changes in the expression level of several surface markers, without affecting the secretion and integrity of small EVs. Our findings lay the foundation for studying the functional role of EVs during early pregnancy, along with the identification of new predictive biomarkers for the severity and outcome of this congenital infection, which are still sorely lacking.

scholarly journals Prenatal Management of Congenital Human Cytomegalovirus Infection in Seropositive Pregnant Patients Treated with Azathioprine

Diagnostics ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 542
Author(s):  
Paolo Ivo Cavoretto ◽  
Chiara Fornara ◽  
Cristina Baldoli ◽  
Alessia Arossa ◽  
Milena Furione ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Human Cytomegalovirus ◽  
Lupus Erythematosus ◽  
Immunosuppressive Agents ◽  
Congenital Infection ◽  
Infectious Agent ◽  
Prenatal Management ◽  
Human Cytomegalovirus Infection ◽  
Low Levels ◽  
Congenital Disabilities

Human cytomegalovirus (HCMV) is the leading infectious agent causing congenital disabilities. The risk of HCMV transmission to the fetus in pregnant women receiving immunosuppressive agents is unknown. We describe two cases of pregnant women with evidence of pre-conception HCMV protective immunity receiving azathioprine for ulcerative colitis or systemic lupus erythematosus. Both women reactivated the HCMV and transmitted the infection to the fetuses. One newborn showed unilateral hearing deficits and brain abnormalities while the other was asymptomatic. The mother of the symptomatic newborn had low levels of total and HCMV-specific blood CD4+ T cells. Women receiving immunosuppressive agents deserve information about the risk of HCMV congenital infection and should be monitored for HCMV infection during pregnancy. Their newborns should be screened for HCMV congenital infection.

Preconceptional Primary Human Cytomegalovirus Infection and Risk of Congenital Infection

2006 ◽  
Vol 193 (6) ◽  
pp. 783-787 ◽  
Author(s):  
Maria Grazia Revello ◽  
Maurizio Zavattoni ◽  
Milena Furione ◽  
Elisa Fabbri ◽  
Giuseppe Gerna
Keyword(s):  
Human Cytomegalovirus ◽  
Cytomegalovirus Infection ◽  
Congenital Infection ◽  
Human Cytomegalovirus Infection

scholarly journals Human Cytomegalovirus modifies placental small extracellular vesicle secretion and composition towards a proviral phenotype to enhance infection of fetal recipient cells

2021 ◽  
Author(s):  
Mathilde Bergamelli ◽  
Hélène Martin ◽  
Yann Aubert ◽  
Jean-Michel Mansuy ◽  
Marlène Marcellin ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Hcmv Infection ◽  
Ex Vivo ◽  
Imaging Techniques ◽  
Fetal Brain ◽  
Congenital Infection ◽  
Protein Composition ◽  
First Trimester ◽  
Ex Vivo Model ◽  
Cytotrophoblast Cell

Although placental small extracellular vesicles (sEVs) are extensively studied in the context of pregnancy, little is known about their role during human cytomegalovirus (hCMV) congenital infection, especially at the beginning of pregnancy. In this study, we examined the consequences of hCMV infection on sEVs production and composition using an immortalized human cytotrophoblast cell line derived from first trimester placenta. By combining complementary approaches of biochemistry, imaging techniques and quantitative proteomic analysis, we showed that hCMV infection increased the yield of sEVs produced by cytotrophoblasts and modified their protein composition towards a proviral phenotype. We further demonstrated that sEVs secreted by hCMV-infected cytotrophoblasts potentiated infection in naive recipient cells of fetal origin, including neural stem cells. Importantly, the enhancement of hCMV infection was also observed with sEVs prepared from either an ex vivo model of infected histocultures from early placenta or from the amniotic fluid of patients naturally infected by hCMV at the beginning of pregnancy. Based on these findings, we propose that placental sEVs could be key actors favoring viral dissemination to the fetal brain during hCMV congenital infection.

scholarly journals Fetal Central Nervous System Derived Extracellular Vesicles: Potential for Non-invasive Tracking of Viral Mediated Fetal Brain Injury

2021 ◽  
Vol 1 ◽  
Author(s):  
Laura Goetzl ◽  
Angela J. Stephens ◽  
Yechiel Schlesinger ◽  
Nune Darbinian ◽  
Nana Merabova ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Brain Injury ◽  
Viral Infection ◽  
Extracellular Vesicles ◽  
First Trimester ◽  
Neurologic Outcome ◽  
Clinical Population ◽  
Non Invasive ◽  
Fetal Infection

Introduction: Extracellular vesicles derived from the fetal central nervous system (FCNSEs) can be purified from maternal serum or plasma using the protein Contactin-2/TAG1that is expressed almost exclusively by developing neurons in the hippocampus, cerebral cortex and cerebellum. We hypothesized that fetal CNSEs could be used to non-invasively detect and quantify viral mediated in-utero brain injury in the first trimester.Materials and Methods: First trimester maternal samples were collected from a human clinical population infected with primary cytomegalovirus (CMV) and a non-human primate model of Zika (ZIKV) infection. In the CMV cohort, a nested case control study was performed comparing pregnancies with and without fetal infection. Cases of fetal infection were further subdivided into those with and without adverse neurologic outcome. ZIKV samples were collected serially following maternal inoculation or saline. All ZIKV cases had histopathologic findings on necropsy. Serum was precipitated with ExoQuick solution and FCEs were isolated with biotinylated anti-Contactin-2/TAG1 antibody-streptavidin matrix immunoabsorption. FCE Synaptopodin (SYNPO) and Neurogranin (NG) protein levels were measured using standard ELISA kits and normalized to the exosome marker CD81.Results: Fetal CNSE SYNPO and NG were significantly reduced in cases of first trimester fetal CMV infection compared to those with infection limited to the mother but could not discriminate between fetal infection with and without adverse neurologic outcome. Following ZIKV inoculation, fetal CNSE SYNPO was reduced by 48 h and significantly reduced by day 4.Discussion: These data are the first to suggest that first trimester non-invasive diagnosis of fetal viral infection is possible. Fetal CNSEs have the potential to augment clinical and pre-clinical studies of perinatal viral infection. Serial sampling may be needed to discriminate between fetuses that are responding to treatment and/or recovering due to innate defenses and those that have ongoing neuronal injury. If confirmed, this technology may advance the paradigm of first trimester prenatal diagnosis and change the calculus for the cost benefit of CMV surveillance programs in pregnancy.

scholarly journals Human Cytomegalovirus Infection of Placental Cytotrophoblasts In Vitro and In Utero: Implications for Transmission and Pathogenesis

2000 ◽  
Vol 74 (15) ◽  
pp. 6808-6820 ◽  
Author(s):  
Susan Fisher ◽  
Olga Genbacev ◽  
Ekaterina Maidji ◽  
Lenore Pereira
Keyword(s):  
Human Cytomegalovirus ◽  
Viral Proteins ◽  
First Trimester ◽  
Maternal Blood ◽  
In Utero ◽  
Uterine Wall ◽  
Chorionic Villi ◽  
Human Cytomegalovirus Infection ◽  
Critical Aspects

ABSTRACT Human cytomegalovirus (CMV) is the leading cause of prenatal viral infection. Affected infants may suffer intrauterine growth retardation and serious neurologic impairment. Analysis of spontaneously aborted conceptuses shows that CMV infects the placenta before the embryo or fetus. In the human hemochorial placenta, maternal blood directly contacts syncytiotrophoblasts that cover chorionic villi and cytotrophoblasts that invade uterine vessels, suggesting possible routes for CMV transmission. To test this hypothesis, we exposed first-trimester chorionic villi and isolated cytotrophoblasts to CMV in vitro. In chorionic villi, syncytiotrophoblasts did not become infected, although clusters of underlying cytotrophoblasts expressed viral proteins. In chorionic villi that were infected with CMV in utero, syncytiotrophoblasts were often spared, whereas cytotrophoblasts and other cells of the villous core expressed viral proteins. Isolated cytotrophoblasts were also permissive for CMV replication in vitro; significantly, infection subsequently impaired the cytotrophoblasts' ability to differentiate and invade. These results suggest two possible routes of CMV transmission to the fetus: (i) across syncytiotrophoblasts with subsequent infection of the underlying cytotrophoblasts and (ii) via invasive cytotrophoblasts within the uterine wall. Furthermore, the observation that CMV infection impairs critical aspects of cytotrophoblast function offers testable hypotheses for explaining the deleterious effects of this virus on pregnancy outcome.

scholarly journals Use of extracellular vesicles from lymphatic drainage as surrogate markers of melanoma progression and BRAFV600E mutation

2019 ◽  
Vol 216 (5) ◽  
pp. 1061-1070 ◽  
Author(s):  
Susana García-Silva ◽  
Alberto Benito-Martín ◽  
Sara Sánchez-Redondo ◽  
Alberto Hernández-Barranco ◽  
Pilar Ximénez-Embún ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Lymphatic Drainage ◽  
Surrogate Markers ◽  
Brafv600e Mutation ◽  
Liquid Biopsies ◽  
Invasive Approach ◽  
Non Invasive ◽  
Melanoma Progression ◽  
Diagnosis And Prognosis ◽  
Patients At Risk

Liquid biopsies from cancer patients have the potential to improve diagnosis and prognosis. The assessment of surrogate markers of tumor progression in circulating extracellular vesicles could be a powerful non-invasive approach in this setting. We have characterized extracellular vesicles purified from the lymphatic drainage also known as exudative seroma (ES) of stage III melanoma patients obtained after lymphadenectomy. Proteomic analysis showed that seroma-derived exosomes are enriched in proteins resembling melanoma progression. In addition, we found that the BRAFV600E mutation can be detected in ES-derived extracellular vesicles and its detection correlated with patients at risk of relapse.

scholarly journals Congenital Human Cytomegalovirus Infection and the Enigma of Maternal Immunity

2017 ◽  
Vol 91 (15) ◽  
Author(s):  
William J. Britt
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Viral Infection ◽  
Human Cytomegalovirus ◽  
Human Fetus ◽  
Cytomegalovirus Infection ◽  
Vaccine Development ◽  
Natural Infection ◽  
Congenital Infection ◽  
Human Cytomegalovirus Infection

ABSTRACT Human cytomegalovirus (HCMV) is the most common viral infection acquired by the developing human fetus and can result in damage to the developing central nervous system. Although vaccine development to modify this congenital infection is ongoing, the unique epidemiology of maternal HCMV infections appears discordant with strategies for vaccine development. Several characteristics of congenital HCMV infections suggest that the efficacy of vaccines designed to induce responses similar to those that follow natural infection will be limited.

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