Detection of rotavirus‐ and norovirus‐specific IgG memory B cells in tonsils

Sharma, Sumit; Hagbom, Marie; Nordgren, Johan; Frodlund, Jonas; Hinkula, Jorma; Ledin, Torbjörn; Svensson, Lennart

doi:10.1002/jmv.25247

Cited by 2 publications

(2 citation statements)

References 23 publications

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“…Of interest, two recent studies have detected diarrheal viruses, including HuNoV, in the nasopharynx of 11.4% [39] and 15.8% [40] of children with acute gastroenteritis, and a prevalence of 0.5% has also been reported for HuNoV RNA in respiratory secretions of children with influenza-like illness [41]. An independent study on subjects without acute gastroenteritis found HuNoV-specific memory B cells in tonsils [42], although they could not detect any marker of viral replication and they did not explore the origin or the role of these cells in that tissue.…”

Section: Discussionmentioning

confidence: 99%

Detection of Norovirus in Saliva Samples from Acute Gastroenteritis Cases and Asymptomatic Subjects: Association with Age and Higher Shedding in Stool

Anfruns-Estrada

Sabrià

Fuentes

et al. 2020

Viruses

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Norovirus infections are a leading cause of acute gastroenteritis outbreaks worldwide and across all age groups, with two main genogroups (GI and GII) infecting humans. The aim of our study was to investigate the occurrence of norovirus in saliva samples from individuals involved in outbreaks of acute gastroenteritis in closed and semiclosed institutions, and its relationship with the virus strain, virus shedding in stool, the occurrence of symptoms, age, and the secretor status of the individual. Epidemiological and clinical information was gathered from norovirus outbreaks occurring in Catalonia, Spain during 2017–2018, and stool and saliva samples were collected from affected and exposed resident individuals and workers. A total of 347 saliva specimens from 25 outbreaks were analyzed. Further, 84% of individuals also provided a paired stool sample. For GII infections, norovirus was detected in 17.9% of saliva samples from symptomatic cases and 5.2% of asymptomatic individuals. Positivity in saliva occurred in both secretors and nonsecretors. None of the individuals infected by norovirus GI was positive for the virus in saliva. Saliva positivity did not correlate with any of the studied symptoms but did correlate with age ≥ 65 years old. Individuals who were positive in saliva showed higher levels of virus shedding in stool. Mean viral load in positive saliva was 3.16 ± 1.08 log10 genome copies/mL, and the predominance of encapsidated genomes was confirmed by propidium monoazide (PMA)xx-viability RTqPCR assay. The detection of norovirus in saliva raises the possibility of oral-to-oral norovirus transmission during the symptomatic phase and, although to a lesser extent, even in cases of asymptomatic infections.

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Section: Discussionmentioning

confidence: 99%

Detection of Norovirus in Saliva Samples from Acute Gastroenteritis Cases and Asymptomatic Subjects: Association with Age and Higher Shedding in Stool

Anfruns-Estrada

Sabrià

Fuentes

et al. 2020

Viruses

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“…The in vitro stimulation strategy is well suited for analysis of lymphoid tissue−resident MBC populations, but limited work has been done to establish protocols that are optimal for unfractionated lymphoid tissue cells (Cao et al, 2010). Tonsillar cell suspensions have been stimulated with a PWM/SAC/CpG cocktail for IgG MBC analysis (Sharma et al, 2019), and our preliminary experiments suggest that R848/IL-2/IL-10 is effective for stimulating IgG MBCs in splenocyte preparations.…”

Section: Commentary Background Informationmentioning

confidence: 98%

Analysis of Antigen‐Specific Human Memory B Cell Populations Based on In Vitro Polyclonal Stimulation

et al. 2020

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Antigen-specific memory B cell (MBC) populations mediate the rapid, strong, and high-affinity secondary antibody responses that play a key role in combating infection and generating protective responses to vaccination. Recently, cell staining with fluorochrome-labeled antigens together with sequencing methods such as Drop-seq and CITE-seq have provided information on the specificity, phenotype, and transcriptome of single MBCs. However, characterization of MBCs at the level of antigen-reactive populations remains an important tool for assessing an individual's B cell immunity and responses to antigen exposure. This is readily performed using a long-established method based on in vitro polyclonal stimulation of MBCs to induce division and differentiation into antibody-secreting cells (ASCs). Post-stimulation antigen-specific measurement of the MBC-derived ASCs (or the secreted antibodies) indicates the size of precursor MBC populations. Additional information about the character of antigen-reactive MBC populations is provided by analysis of MBC-derived antibodies of particular specificities for binding avidity and functionality. This article outlines a simple and reliable strategy for efficient in vitro MBC stimulation and use of the ELISpot assay as a post-stimulation readout to determine the size of antigen-specific MBC populations. Other applications of the in vitro stimulation technique for MBC analysis are discussed. The following protocols are included.

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Detection of rotavirus‐ and norovirus‐specific IgG memory B cells in tonsils

Cited by 2 publications

References 23 publications

Detection of Norovirus in Saliva Samples from Acute Gastroenteritis Cases and Asymptomatic Subjects: Association with Age and Higher Shedding in Stool

Detection of Norovirus in Saliva Samples from Acute Gastroenteritis Cases and Asymptomatic Subjects: Association with Age and Higher Shedding in Stool

Analysis of Antigen‐Specific Human Memory B Cell Populations Based on In Vitro Polyclonal Stimulation

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