Recombinant Echovirus E3 VP1 Protein [His]

Background: Echovirus (E) 30 (E-30) meningitis is characterized by neuroinflammation involving immune cell pleocytosis at the protective barriers of the central nervous system (CNS). In this context, infection of the blood-cerebrospinal fluid barrier (BCSFB), which has been demonstrated to be involved in enteroviral CNS pathogenesis, may affect the tight junction (TJ) and adherens junction (AJ) function and morphology. Methods: We used an in vitro human choroid plexus epithelial (HIBCPP) cell model to investigate the effect of three clinical outbreak strains (13-311, 13-759, and 14-397) isolated in Germany in 2013, and compared them to E-30 Bastianni. Conducting transepithelial electrical resistance (TEER), paracellular dextran flux measurement, quantitative real-time polymerase chain reaction (qPCR), western blot, and immunofluorescence analysis, we investigated TJ and AJ function and morphology as well as strain-specific E-30 infection patterns. Additionally, transmission electron and focused ion beam microscopy electron microscopy (FIB-SEM) was used to evaluate the mode of leukocyte transmigration. Genome sequencing and phylogenetic analyses were performed to discriminate potential genetic differences among the outbreak strains. Results: We observed a significant strain-dependent decrease in TEER with strains E-30 Bastianni and 13-311, whereas paracellular dextran flux was only affected by E-30 Bastianni. Despite strong similarities among the outbreak strains in replication characteristics and particle distribution, strain 13-311 was the only outbreak isolate revealing comparable disruptive effects on TJ (Zonula Occludens (ZO) 1 and occludin) and AJ (E-cadherin) morphology to E-30 Bastianni. Notwithstanding significant junctional alterations upon E-30 infection, we observed both para-and transcellular leukocyte migration across HIBCPP cells. Complete genome sequencing revealed differences between the strains analyzed, but no explicit correlation with the observed strain-dependent effects on HIBCPP cells was possible. Conclusion: The findings revealed distinct E-30 strain-specific effects on barrier integrity and junctional morphology. Despite E-30-induced barrier alterations leukocyte trafficking did not exclusively occur via the paracellular route.

Objective: Evaluation of hand, foot, and mouth disease (HFMD) diagnostic strategies in pregnancy and the risk of HFMD-related fetopathy. Study design: Pregnant women consecutively evaluated between 2010 and 2016 at the Tuscany Reference Center for Infectious Diseases in Pregnancy for HFMD were enrolled. A descriptive analysis of infected patients/newborns data and literature review were carried out. Result: Of the 128 women evaluated, 52 (41%) were symptomatic: 32 (61.5%) developed HFM vesicles, 12 (23%) palmoplantar vesicles, and 8 (15.5%) oral aphthae. Serological positivity and direct Enterovirus detection on blood and vesicle were obtained in 1.9% (1/52), 9.1% (1/11), and 68.7% (11/16), respectively. Three miscarriage and few cases of fetal/neonatal anomalies were reported. Conclusion: HFMD diagnosis is primarily a clinical diagnosis. Direct viral detection is more sensitive than serology. Considering our series and literature review, data on embryo-fetal-neonatal outcomes are not conclusive. Although the role of EV as causative agents of congenital defects remains uncertain, the described cases of unfavorable outcome impose prudence and monitoring of pregnant women with HFMD throughout the gestation.