Differential permissivity of human cerebrovascular endothelial cells to enterovirus infection and specificities of enterovirus 71 in crossing an in vitro model of human blood brain barrier

Human cerebral microvascular endothelial cells (hCMEC/D3 cell line) form a steady polarized barrier when cultured in vitro on a permeable membrane. Their susceptibility to enterovirus (EV) strains was analysed to investigate how these viruses may cross the blood-brain barrier. A sample of 88 virus strains was selected on phylogenetic features among 44 epidemiologically relevant types of the four EV species A-D. The EV-A71 genome was replicated at substantial rates while the infectious virus was released at extremely low but sustained rates at both barrier sides for at least 4 days. EV-A71 antigens were detected in a limited number of cells. The properties of the endothelial barrier (structure and permeability) remained intact throughout infection. The chronic EV-A71 infection was in sharp contrast with the productive infection of cytolytic EVs (e.g. echoviruses 6 and 30). The hCMEC/D3 barriers infected with the latter EVs exhibited elevated proportions of apoptotic and necrotic cells, which resulted in major injuries to the endothelial barriers with dramatic increase of paracellular permeability and virus crossing to the abluminal side. The following intracellular rearrangements were also seen: early destruction of the actin cytoskeleton, remodelling of intracellular membranes, and reorganization of the mitochondrion network in a small cluster near the perinuclear space

Recherche de virus de l'hépatite C recombinants chez des patients multi-exposés

Depuis 2002, 5 virus recombinants inter-génotypiques du virus de l'hépatite C (RF_2k/1b, RF_2b/1b, RF_2i/6p, RF-2/5 et RF_2b/6w) ont été mis en évidence chez des patients toxicomanes ou hémophiles. Le génome des virus recombinants inter-génotypiques appartient au génotype 2 en 5' et à un autre génotype en 3'. Le point de recombinaison entre les 2 génotypes se situe dans la région génomique NS2-NS3. La fréquence de survenue des recombinaisons n'est pas encore évaluée car les techniques de génotypage actuellement utilisées en pratique courante, basées l'analyse d'une seule région génomique, ne permettent pas de les détecter. Nous avons recherché des souches recombinantes de VHC dans une population de 63 patients atteints d'hépatite chronique C, usagers de drogues par voie intra-veineuse ou hémophiles. La recherche a été réalisée par séquençage puis analyse phylogénétique de 2 régions génomiques différentes, l'une en 5' du génome codant pour la protéine d'enveloppe E1, et l'autre en 3' codant pour la polymérase virale NS5B. La comparaison des analyses réalisées sur les 2 extrémités du génome n'a permis de mettre en évidence ni recombinants inter-génotypiques ni cas de co-infection dans la population étudiée. En dépit d'une prévalence qui parait actuellement faible, il parait nécessaire d'étudier les virus recombinants inter-génotypiques en recherche fondamentale et clinique afin de déterminer leur mécanisme de formation, leur épidémiologie mondiale, leur prévalence et surtout leur sensibilité aux traitements actuels et du futur. Leur diffusion pourrait amener à reconsidérer l'utilisation en pratique courante des techniques de génotypage basées sur l'analyse d'une seule région du génome.CLERMONT FD-BCIU-Santé (631132104) / SudocLYON1-BU Santé (693882101) / SudocSudocFranceF

Evolutionary history of hepatitis C virus genotype 5a in France, a multicenter ANRS study

The epidemic history of HCV genotype 5a is poorly documented in France, where its prevalence is very low, except in a small central area, where it accounts for 14.2% of chronic hepatitis C cases. A Bayesian coalescent phylogenetic investigation based on the E1 envelope gene and a non-structural genomic segment (NS3/4) was carried out to trace the origin of this epidemic using a large sample of genotype 5a isolates collected throughout France. The dates of documented transmissions by blood transfusion were used to calibrate five nodes in the phylogeny. The results of the E1 gene analysis showed that the best-fitting population dynamic model was the expansion growth model under a relaxed molecular clock. The rate of nucleotide substitutions and time to the most recent common ancestors (tMRCA) of genotype 5a isolates were estimated. The divergence of all the French HCV genotype 5a strains included in this study was dated to 1939 [95% HPD: 1921-1956], and the tMRCA of isolates from central France was dated to 1954 [1942-1967], which is in agreement with epidemiological data. NS3/4 analysis provided similar estimates with strongly overlapping HPD values. Phylodynamic analyses give a plausible reconstruction of the evolutionary history of HCV genotype 5a in France, suggesting the concomitant roles of transfusion, iatrogenic route and intra-familial transmission in viral diffusion. © 2011 Elsevier B.V.0SCOPUS: ar.jSCOPUS: ar.jinfo:eu-repo/semantics/publishe

Nearly Complete Genome Sequence of Enterovirus Type A119 from Sewage in France in 2015

International audienceA nearly complete genome sequence of enterovirus type A119 was determined from an urban wastewater sample collected during a surveillance campaign in 2015 in Clermont-Ferrand, France. The partial VP1 sequence is a close relative of other partial enterovirus type A119 sequences detected in France and South Africa in the same year

Complete Genome Sequence of an Enterovirus Type C116 Strain Recovered from Urban Sewage and Determined by Deep Sequencing

International audienceWastewater surveillance allowed the detection of an enterovirus (EV) type rarely reported in clinical settings. We detected an EV-C116 strain in a wastewater sample in France and characterized its complete genome. This virus was genetically closely related to African strains but distantly related to the only complete genome previously described

Genomic Variations in Echovirus 30 Persistent Isolates Recovered from a Chronically Infected Immunodeficient Child and Comparison with the Reference Strain

Seven sequential isolates of echovirus type 30 (EV30) were recovered over 22 months from a child with severe combined immune deficiency syndrome. The nucleotide sequences of the 5′ halves of the genomes (4,400 nucleotides) of the first (S1) and last (S7) isolates were determined and compared with that of the EV30 Bastianni reference strain, also determined in this study. In genome regions P1 and P2, 101 variations were identified between the two isolates. Synonymous differences far outnumbered nonsynonymous differences. Amino acid changes affected both capsid and nonstructural polypeptides (particularly 2B). The VP1 nucleotide sequences of the seven isolates were determined to analyze genome evolution during the chronic infection. In the phylogenetic tree, the seven isolates were directly related to the prototype strain in an individual monophyletic group, strongly suggesting that the chronic infection in the child arose from a single persistent EV30 isolate. Four lineages were observed in the persistent isolates. Isolates S2, S4, S5, and S6 were close relatives of one another, whereas isolates S1 and S3 formed individual lineages. Isolate S7, distantly related to all other isolates, formed the fourth lineage. These findings suggest the quasispecies nature of the genomes of the seven sequential EV30 isolates. Grouping of persistent isolates on the basis of replicative capacities was consistent with phylogenetic relationships. Overall, the results indicate that genetically related EV30 variants with different replicative capacities coexisted in a carrier state, probably in the gastrointestinal tract, during the infection of the child

Monitoring of enterovirus diversity in wastewater by ultra-deep sequencing: An effective complementary tool for clinical enterovirus surveillance

International audienceIn a one-year (October 2014eOctober 2015) pilot study, we assessed wastewater monitoring with sustained sampling for analysis of global enterovirus (EV) infections in an urban community. Wastewater was analysed by ultra-deep sequencing (UDS) after PCR amplification of the partial VP1 capsid protein gene. The nucleotide sequence analysis showed an unprecedented diversity of 48 EV types within the community, which were assigned to the taxonomic species A (n ¼ 13), B (n ¼ 23), and C (n ¼ 12). During the same period, 26 EV types, of which 22 were detected in wastewater, were identified in patients referred to the teaching hospital serving the same urban population. Wastewater surveillance detected a silent circulation of 26 EV types including viruses reported in clinically rare respiratory diseases. Wastewater monitoring as a supplementary procedure can complement clinical surveillance of severe diseases related to non-polio EVs and contribute to the final stages of poliomyelitis eradication

Enterovirus meningitis in Mayotte French Comoros Island, March-June 2019

International audienc