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Title: Human astrovirus MLB replication in vitro: persistence in extraintestinal cell lines
Author: Vu Cantero, Diem-Lan
Bosch, Albert
Pintó Solé, Rosa María
Ribes Mora, Enric
Guix Arnau, Susana
Keywords: Malalties intestinals
Intestinal diseases
Issue Date: 14-Jun-2019
Publisher: American Society for Microbiology
Abstract: MLB astroviruses were identified 10 years ago in feces from children with gastroenteritis of unknown etiology and have been unexpectedly detected in severe cases of meningitis/encephalitis, febrile illness of unknown etiology, and respiratory syndromes. The aim of this study was to establish a cell culture system supporting MLB astrovirus replication. We used two clinical strains to infect several cell lines, an MLB1 strain from a gastroenteritis case, and an MLB2 strain associated with a neurologic infection. Efforts to propagate the viruses in the Caco-2 cell line were unsuccessful. In contrast, we identified two human nonintestinal cell lines, Huh-7 and A549, permissive for both genotypes. After serial passages in the Huh-7.5 cell line, the adapted strains were able to establish persistent infections in the Huh-7.5, Huh-7AI, and A549 cell lines, with high viral loads (up to 10 log10 genome copies/ml) detected by quantitative reverse transcription-PCR (RT-qPCR) in the culture supernatant. Immunofluorescence assays demonstrated infection in about 10% of cells in persistently infected cultures. Electron microscopy revealed particles of 32 to 33 nm in diameter after negative staining of cell supernatants and capsid arrays in ultrathin sections with a particularly high production in Huh-7.5 cells. Interferon (IFN) expression by infected cells and effect of exogenous IFN varied depending on the type of infection and the cell line. The availability of a cell culture system to propagate MLB astroviruses represents a key step to better understand their replicative cycle, as well as a source of viruses to conduct a wide variety of basic virologic studies
Note: Versió postprint del document publicat a:
It is part of: Journal of Virology, 2019, vol. 93, num. 13
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ISSN: 0022-538X
Appears in Collections:Articles publicats en revistes (Genètica, Microbiologia i Estadística)

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