Disrupted Peyer’s Patch Microanatomy in COVID-19 Including Germinal Centre Atrophy Independent of Local Virus
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Abstract
Confirmed SARS-coronavirus-2 infection with gastrointestinal symptoms and changes in microbiota associated with coronavirus disease 2019 (COVID-19) severity have been previously reported, but the disease impact on the architecture and cellularity of ileal Peyer’s patches (PP) remains unknown. Here we analysed post-mortem tissues from throughout the gastrointestinal (GI) tract of patients who died with COVID-19. When virus was detected by PCR in the GI tract, immunohistochemistry identified virus in epithelium and lamina propria macrophages, but not in lymphoid tissues. Immunohistochemistry and imaging mass cytometry (IMC) analysis of ileal PP revealed depletion of germinal centres (GC), disruption of B cell/T cell zonation and decreased potential B and T cell interaction and lower nuclear density in COVID-19 patients. This occurred independent of the local viral levels. The changes in PP demonstrate that the ability to mount an intestinal immune response is compromised in severe COVID-19, which could contribute to observed dysbiosis.
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SciScore for 10.1101/2021.12.17.473179: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis The tissues were visualized under a light microscope and after choosing a region of interest (where the lymphoid tissue was present), data was acquired on a Hyperion imaging system coupled to a Helios Mass Cytometer (Fluidigm) tuned with 3 element tuning slide, at a laser frequency mean of 200Hz and 6dB power. Table 2: Resources
Antibodies Sentences Resources The blocking, staining with primary and secondary antibodies were realized according to the manufactures of the following IHC kits: Mouse and rabbit specific HRP/DAB IHC detection Kit micro-polymer (Abcam, Cat.ab236466) and … SciScore for 10.1101/2021.12.17.473179: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis The tissues were visualized under a light microscope and after choosing a region of interest (where the lymphoid tissue was present), data was acquired on a Hyperion imaging system coupled to a Helios Mass Cytometer (Fluidigm) tuned with 3 element tuning slide, at a laser frequency mean of 200Hz and 6dB power. Table 2: Resources
Antibodies Sentences Resources The blocking, staining with primary and secondary antibodies were realized according to the manufactures of the following IHC kits: Mouse and rabbit specific HRP/DAB IHC detection Kit micro-polymer (Abcam, Cat.ab236466) and ImmPRESS[R] Duet double staining kit anti-rabbit AP/anti-mouse HRP (Vector laboratories, Cat.mp7724). anti-rabbit AP/anti-mouse HRPsuggested: NoneAnti-SARS-CoV-2 spike antibody, targeting the S2 subunit, and anti-SARS-CoV-2 nucleocapsid (1:300) antibody were acquired from Insight Biotechnology (Cat. GTX632604) and BioserverUK (Cat.BSV- COV-AB-13), respectively. Anti-SARS-CoV-2suggested: NoneS2 subunit,suggested: Noneanti-SARS-CoV-2 nucleocapsidsuggested: NoneSoftware and Algorithms Sentences Resources The data was normalized in GraphPad Prism software v9.0 GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)The % of area occupied by the follicle was determined in Image J (version 1.0 for Mac OS X) using the image selected in R studio. Image Jsuggested: (ImageJ, RRID:SCR_003070)(GraphPad Software, Inc., La Jolla, CA). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your code.
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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