Detection of respiratory viruses in primary cholesteatoma tissues

Viana, Rosa Maria Mendes; Souza, Juliano P.; Jorge, Daniel Macedo de Melo; Martins, Ronaldo B.; Castro, Ítalo A.; Cardoso, Ricardo; Volpini, Lays Paula Bondi; Luna, Luciano Kleber de Souza; Spano, Liliana Cruz; Bellei, Nancy; Chahud, Fernando; Arruda, Eurico; Hyppolito, Miguel Ângelo

doi:10.1002/jmv.27107

Cited by 2 publications

(2 citation statements)

References 24 publications

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“…S1 ). Tissue sections (3 µm) were tested as previously published ( 9 ). Slides were stained with primary monoclonal rabbit antibody anti-SARS-CoV-2 nucleocapsid (Creative Diagnostics #CABT-RMJ1) or with a rabbit anti-SARS-CoV-2 NSP-16 (Abcam, ab284038).…”

Section: Methodsmentioning

confidence: 99%

Tonsils are major sites of persistence of SARS-CoV-2 in children

Lima,

Martins,

Miura

et al. 2023

Microbiol Spectr

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In the present study, we show that SARS-CoV-2 can infect palatine tonsils, adenoids, and secretions in children without symptoms of COVID-19, with no history of recent upper airway infection. We studied 48 children undergoing tonsillectomy due to snoring/OSA or recurrent tonsillitis between October 2020 and September 2021. Nasal cytobrushes, nasal washes, and tonsillar tissue fragments obtained at surgery were tested by RT-qPCR, immunohistochemistry (IHC), flow cytometry, and neutralization assay. We detected the presence of SARS-CoV-2 in at least one specimen tested in 27% of patients. IHC revealed the presence of the viral nucleoprotein in epithelial surface and in lymphoid cells in both extrafollicular and follicular regions, in adenoids and palatine tonsils. Also, IHC for the SARS-CoV-2 non-structural protein NSP-16 indicated the presence of viral replication in 53.8% of the SARS-CoV-2-infected tissues. Flow cytometry showed that CD20 + B lymphocytes were the most infected phenotypes, followed by CD4+ lymphocytes and CD123 dendritic cells, CD8+ T lymphocytes, and CD14+ macrophages. Additionally, IF indicated that infected tonsillar tissues had increased expression of ACE2 and TMPRSS2. NGS sequencing demonstrated the presence of different SARS-CoV-2 variants in tonsils from different tissues. SARS-CoV-2 antigen detection was not restricted to tonsils but was also detected in nasal cells from the olfactory region. Palatine tonsils and adenoids are sites of prolonged RNA presence by SARS-CoV-2 in children, even without COVID-19 symptoms. IMPORTANCE This study shows that SRS-CoV-2 of different lineages can infect tonsils and adenoids in one quarter of children undergoing tonsillectomy. These findings bring advancement to the area of SARS-CoV-2 pathogenesis, by showing that tonsils may be sites of prolonged infection, even without evidence of recent COVID-19 symptoms. SARS-CoV-2 infection of B and T lymphocytes, macrophages, and dendritic cells may interfere with the mounting of immune responses in these secondary lymphoid organs. Moreover, the shedding of SARS-CoV-2 RNA in respiratory secretions from silently infected children raises concern about possible diagnostic confusion in the presence of symptoms of acute respiratory infections caused by other etiologies.

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Section: Methodsmentioning

confidence: 99%

Tonsils are major sites of persistence of SARS-CoV-2 in children

Lima,

Martins,

Miura

et al. 2023

Microbiol Spectr

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show abstract

“…Tissue sections (3µm) were deparaffinized, rehydrated and subjected to the previously published protocols. 8 Red-purple signal was developed with AEC peroxidase system kit (SK-4800, Vector Laboratories, Burlingame, CA) and sections were counterstained with Harris hematoxylin (Vector) and scanned with a ScanScope VS120 (Olympus Life Sciences, Tokyo, Japan) in bright field, using 400× magnification.…”

Section: Immunohistochemistry For Sars-cov-2 Antigen In Adenotonsilla...mentioning

confidence: 99%

Tonsils Are Major Sites of Prolonged Sars-Cov-2 Infection in Children

Melquiades

Martins

Miura

et al. 2023

Preprint

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In the present study, we show that SARS-CoV-2 can infect palatine tonsils and adenoids in children without symptoms of COVID-19, with no history of recent upper airway infection. We studied 48 children undergoing tonsillectomy due to snoring/OSA or recurrent tonsillitis between October 2020 and September 2021. Briefly, nasal cytobrush (NC), nasal wash (NW) and tonsillar tissue fragments obtained at surgery were tested by RT-PCR, immunohistochemistry (IHC), flow cytometry and neutralization assay. We detected the presence of SARS-CoV-2 in at least one specimen tested in 25% of patients (20% in palatine tonsils and 16.27% in adenoids, 10.41% of NC and 6.25% of NW). Importantly, in 2 of the children there was evidence of laboratory-confirmed acute infection 2 and 5 months before surgery. IHC revealed the presence of SARS-CoV-2 nucleoprotein in epithelial surface and in lymphoid cells in both extrafollicular and follicular regions, in adenoids and palatine tonsils. Flow cytometry showed that CD20+B lymphocytes were the most infected phenotypes by SARS-CoV-2 NP, followed by CD4+ and CD8+ T lymphocytes, and CD14+ macrophages and dendritic cells. Additionally, IF indicated that SARS-CoV-2-infected tonsillar tissues had increased expression of ACE2 and TMPRSS2. NGS sequencing demonstrated the presence of different SARS CoV-2 variants in tonsils from different tissues. SARS-CoV-2 antigen detection was not restricted to tonsils, but was also detected in nasal cells from the olfactory region. In conclusion, palatine tonsils and adenoids are sites of prolonged infection by SARS-CoV-2 in children, even without COVID-19 symptoms.

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Detection of respiratory viruses in primary cholesteatoma tissues

Cited by 2 publications

References 24 publications

Tonsils are major sites of persistence of SARS-CoV-2 in children

Tonsils are major sites of persistence of SARS-CoV-2 in children

Tonsils Are Major Sites of Prolonged Sars-Cov-2 Infection in Children

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