Quantifying Human Norovirus Virus-like Particles Binding to Commensal Bacteria Using Flow Cytometry

Madrigal, Jasmine L.; Jones, Melissa K.

doi:10.3791/61048

Cited by 8 publications

(8 citation statements)

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“…To generate stationary phase cultures bacteria were inoculated into appropriate medium and incubated overnight at 37 °C [ 20 ]. E. cloacae , E. coli , and P. aeruginosa were grown with shaking (200 rpm) while L acidophilus and L. gasseri were grown statically.…”

Section: Methodsmentioning

confidence: 99%

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Attach Me If You Can: Murine Norovirus Binds to Commensal Bacteria and Fungi

Madrigal

Bhar

Hackett

et al. 2020

Viruses

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The presence of commensal bacteria enhances both acute and persistent infection of murine noroviruses. For several enteric viral pathogens, mechanisms by which these bacteria enhance infection involve direct interactions between the virus and bacteria. While it has been demonstrated that human noroviruses bind to a variety of commensal bacteria, it is not known if this is also true for murine noroviruses. The goal of this study was to characterize interactions between murine noroviruses and commensal bacteria and determine the impact of bacterial growth conditions, incubation temperature and time, on murine norovirus attachment to microbes that comprise the mammalian microbiome. We show that murine noroviruses bind directly to commensal bacteria and show similar patterns of attachment as human norovirus VLPs examined under the same conditions. Furthermore, while binding levels are not impacted by the growth phase of the bacteria, they do change with time and incubation temperature. We also found that murine norovirus can bind to a commensal fungal species, Candida albicans.

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Section: Methodsmentioning

confidence: 99%

“…HNoV attachment was quantified by flow cytometry as previously described [ 20 ]. MNV attachment was quantified using RT-qPCR.…”

Section: Methodsmentioning

confidence: 99%

Attach Me If You Can: Murine Norovirus Binds to Commensal Bacteria and Fungi

Madrigal

Bhar

Hackett

et al. 2020

Viruses

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“…For detecting RV-binding bacteria, we followed a strategy similar to that recently described to quantify the in vitro binding of noroviral-virus-like particles to gut commensal bacteria [41]. Two-hundred milligrams of each stool sample was suspended in 1 mL of 0.9% saline solution (SS) by pipetting, followed by 10 s vortexing.…”

Section: Preparation Of Microbiota Samples From Stools and Cell Sortingmentioning

confidence: 99%

Interaction of Intestinal Bacteria with Human Rotavirus during Infection in Children

Gozalbo-Rovira

Rubio‐del‐Campo

Santiso-Bellón

et al. 2021

IJMS

184

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The gut microbiota has emerged as a key factor in the pathogenesis of intestinal viruses, including enteroviruses, noroviruses and rotaviruses (RVs), where stimulatory and inhibitory effects on infectivity have been reported. With the aim of determining whether members of the microbiota interact with RVs during infection, a combination of anti-RV antibody labeling, fluorescence-activated cell sorting and 16S rRNA amplicon sequencing was used to characterize the interaction between specific bacteria and RV in stool samples of children suffering from diarrhea produced by G1P[8] RV. The genera Ruminococcus and Oxalobacter were identified as RV binders in stools, displaying enrichments between 4.8- and 5.4-fold compared to samples nonlabeled with anti-RV antibodies. In vitro binding of the G1P[8] Wa human RV strain to two Ruminococcus gauvreauii human isolates was confirmed by fluorescence microscopy. Analysis in R. gauvreauii with antibodies directed to several histo-blood group antigens (HBGAs) indicated that these bacteria express HBGA-like substances on their surfaces, which can be the target for RV binding. Furthermore, in vitro infection of the Wa strain in differentiated Caco-2 cells was significantly reduced by incubation with R. gauvreauii. These data, together with previous findings showing a negative correlation between Ruminococcus levels and antibody titers to RV in healthy individuals, suggest a pivotal interaction between this bacterial group and human RV. These results reveal likely mechanisms of how specific bacterial taxa of the intestinal microbiota could negatively affect RV infection and open new possibilities for antiviral strategies.

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“…For detecting RV-binding bacteria we followed a strategy similar to that recently described to quantify the in vitro binding of noroviral virus-like particles to gut commensal bacteria [41]. Two-hundred mg of each stool sample were suspended in 1 mL of 0.9% saline solution (SS) by pipetting, followed by 10 seconds vortexing.…”

Section: Methodsmentioning

confidence: 99%

Interaction of intestinal bacteria with human rotavirus during infection in children

Gozalbo-Rovira

Rubio‐del‐Campo

Santiso-Bellón

et al. 2020

Preprint

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The gut microbiota has emerged as a key factor in the pathogenesis of intestinal viruses, including enteroviruses, noroviruses and rotaviruses (RV), where stimulatory and inhibitory effects on infectivity have been reported. With the aim of determining whether members of the microbiota interact with RV during infection, a combination of anti-RV antibody labelling, fluorescence-activated cell sorting and 16S rRNA amplicon sequencing was used to characterize the interaction between specific bacteria and RV in stool samples of children suffering diarrhea produced by G1P[8] RV. The genera Ruminococcus and Oxalobacter were identified as RV binders in stools, displaying enrichments between 4.8 to 5.4-fold compared to samples non-labelled with anti-RV antibodies. In vitro binding of the G1P[8] Wa human RV strain to two Ruminococcus gauvreauii human isolates was confirmed by fluorescence microscopy. Analysis in R. gauvreauii with antibodies directed to several histo-blood group antigens (HBGA) indicated that these bacteria express HBGA-like substances at their surfaces that can be the target for RV binding. Furthermore, in vitro infection of Wa strain in differentiated Caco-2 cells was significantly reduced by incubation with R. gauvreauii. These data, together with previous findings that had shown a negative correlation between Ruminococcus levels and antibody titers to RV in healthy individuals, suggest a pivotal interaction between this bacterial group and human RV. These results reveal likely mechanisms on how specific bacterial taxa of the intestinal microbiota could negatively affect RV infection and open new possibilities for anti-viral strategies.

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Quantifying Human Norovirus Virus-like Particles Binding to Commensal Bacteria Using Flow Cytometry

Cited by 8 publications

References 0 publications

Attach Me If You Can: Murine Norovirus Binds to Commensal Bacteria and Fungi

Attach Me If You Can: Murine Norovirus Binds to Commensal Bacteria and Fungi

Interaction of Intestinal Bacteria with Human Rotavirus during Infection in Children

Interaction of intestinal bacteria with human rotavirus during infection in children

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