ef39182984f8498fd569b8d84fd19c8b pmc_9265332.pdf 353ff8ab75b6df87d4d7b45d759c50d54e98a24f pmc_9265332.pdf fb3534277266d6275cf4bb26f9e362fbe653c6b02b19959d713143a6427382c3 pmc_9265332.pdf Title: Mucosal Plasma Cell Activation and Proximity to Nerve Fibres Are Associated with Glycocalyx Reduction in Diarrhoea-Predominant Irritable Bowel Syndrome: Jejunal Barrier Alterations Underlying Clinical Manifestations Subject: Irritable bowel syndrome (IBS) is a disorder of brain-gut interaction characterised by abdominal pain and changes in bowel habits. In the diarrhoea subtype (IBS-D), altered epithelial barrier and mucosal immune activation are associated with clinical manifestations. We aimed to further evaluate plasma cells and epithelial integrity to gain understanding of IBS-D pathophysiology. One mucosal jejunal biopsy and one stool sample were obtained from healthy controls and IBS-D patients. Gastrointestinal symptoms, stress, and depression scores were recorded. In the jejunal mucosa, RNAseq and gene set enrichment analyses were performed. A morphometric analysis by electron microscopy quantified plasma cell activation and proximity to enteric nerves and glycocalyx thickness. Immunoglobulins concentration was assessed in the stool. IBS-D patients showed differential expression of humoral pathways compared to controls. Activation and proximity of plasma cells to nerves and IgG concentration were also higher in IBS-D. Glycocalyx thickness was lower in IBS-D compared to controls, and this reduction correlated with plasma cell activation, proximity to nerves, and clinical symptoms. These results support humoral activity and loss of epithelial integrity as important contributors to gut dysfunction and clinical manifestations in IBS-D. Additional studies are needed to identify the triggers of these alterations to better define IBS-D pathophysiology. Keywords: intestinal plasma cells; intestinal glycocalyx; IBS-D; mucosal ultrastructure; intestinal barrier dysfunction; mucosal nerve fibres Author: Cristina Pardo-Camacho, John-Peter Ganda Mall, Cristina Martínez, Marc Pigrau, Elba Expósito, Mercé Albert-Bayo, Elisa Melón-Ardanaz, Adoración Nieto, Bruno Rodiño-Janeiro, Marina Fortea, Danila Guagnozzi, Amanda Rodriguez-Urrutia, Inés de Torres, Ignacio Santos-Briones, Fernando Azpiroz, Beatriz Lobo, Carmen Alonso-Cotoner, Javier Santos, Ana M. González-Castro and Maria Vicario Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Tue Jun 28 12:55:07 2022 CEST ModDate: Tue Jun 28 13:02:21 2022 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2124486 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 MWHIBA+VnURWPalladioL Type 1 Custom yes yes no 15 0 CKQKMF+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 WZXFBQ+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 QHGNSK+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 PFKOHU+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 46 0 JHXMEH+CMSY10 Type 1 Builtin yes yes no 71 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 78 0 OVNEXW+EURM10 Type 1 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Assessment Destination: subsection.2.1 Item: Title: Experimental Design and Procedures Destination: subsection.2.2 Item: Title: Collection of Biological Samples Destination: subsection.2.3 Item: Title: Analytical Procedures Destination: subsection.2.4 Children: Item: Title: Gene Expression Analysis by RNAseq and Q-RT-PCR Destination: subsubsection.2.4.1 Item: Title: Histological Assessment Destination: subsubsection.2.4.2 Item: Title: Ultrastructural Analysis Destination: subsubsection.2.4.3 Item: Title: Immunoglobulin Quantification Destination: subsubsection.2.4.4 Item: Title: Statistical Analysis Destination: subsection.2.5 Item: Title: Results Destination: section.3 Children: Item: Title: Study Population Destination: subsection.3.1 Item: Title: The Jejunal Mucosa Shows Higher Representation of Humoral Activity Pathways in IBS-D Compared to Health Destination: subsection.3.2 Item: Title: Ultrastructural Assessment Shows Differential Plasma Cell Activation in the Jejunal Mucosa in IBS-D Destination: subsection.3.3 Item: Title: Reduced Glycocalyx in IBS-D in Association with Plasma Cell Activation Destination: subsection.3.4 Item: Title: Increased Mucosal Plasma Cell Proximity to Nerve Fibres in IBS-D, in Association with Plasma cell Activation and Glycocalyx Reduction Destination: subsection.3.5 Item: Title: Higher Concentration of IgG in the Intestinal Lumen in IBS-D Destination: subsection.3.6 Item: Title: Associations between Clinical, Histological, and Biological Variables Destination: subsection.3.7 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Mucosal Plasma Cell Activation and Proximity to Nerve Fibres Are Associated with Glycocalyx Reduction in Diarrhoea-Predominant Irritable Bowel Syndrome: Jejunal Barrier Alterations Underlying Clinical Manifestations Author: Cristina Pardo-Camacho, John-Peter Ganda Mall, Cristina Martínez, Marc Pigrau, Elba Expósito, Mercé Albert-Bayo, Elisa Melón-Ardanaz, Adoración Nieto, Bruno Rodiño-Janeiro, Marina Fortea, Danila Guagnozzi, Amanda Rodriguez-Urrutia, Inés de Torres, Ignacio Santos-Briones, Fernando Azpiroz, Beatriz Lobo, Carmen Alonso-Cotoner, Javier Santos, Ana M. González-Castro and Maria Vicario Subject: Irritable bowel syndrome (IBS) is a disorder of brain-gut interaction characterised by abdominal pain and changes in bowel habits. In the diarrhoea subtype (IBS-D), altered epithelial barrier and mucosal immune activation are associated with clinical manifestations. We aimed to further evaluate plasma cells and epithelial integrity to gain understanding of IBS-D pathophysiology. One mucosal jejunal biopsy and one stool sample were obtained from healthy controls and IBS-D patients. Gastrointestinal symptoms, stress, and depression scores were recorded. In the jejunal mucosa, RNAseq and gene set enrichment analyses were performed. A morphometric analysis by electron microscopy quantified plasma cell activation and proximity to enteric nerves and glycocalyx thickness. Immunoglobulins concentration was assessed in the stool. IBS-D patients showed differential expression of humoral pathways compared to controls. Activation and proximity of plasma cells to nerves and IgG concentration were also higher in IBS-D. Glycocalyx thickness was lower in IBS-D compared to controls, and this reduction correlated with plasma cell activation, proximity to nerves, and clinical symptoms. These results support humoral activity and loss of epithelial integrity as important contributors to gut dysfunction and clinical manifestations in IBS-D. Additional studies are needed to identify the triggers of these alterations to better define IBS-D pathophysiology. 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