scholarly journals In vivo gluten challenge in coeliac disease

2001 ◽  
Vol 15 (4) ◽  
pp. 243-247 ◽  
Author(s):  
HJ Ellis ◽  
PJ Ciclitira
Keyword(s):  
Celiac Disease ◽  
Invasive Procedure ◽  
Intraepithelial Lymphocytes ◽  
Wheat Starch ◽  
Intestinal Biopsy ◽  
Jejunal Biopsy ◽  
Small Intestinal Biopsy ◽  
Immunohistochemical Techniques

In vivo gluten challenge has been used since the early 1950s to study the role of cereal fractions in celiac disease. While early studies relied on crude indicators of celiac toxicity, the advent of jejunal biopsy and sophisticated immunohistochemical techniques has allowed accurate studies to be performed. Studies to determine the nature of the cereal component that is toxic to patients with celiac disease have concentrated on wheat because of its nutritional importance. A number of in vitro studies indicated the presence of one or more celiac-activating epitopes with theN-terminus of the A-gliadin molecule. In vivo challenge with three synthetic peptides subsequently indicated the toxicity of a peptide corresponding to amino acids 31 to 49 of A-gliadin. In vivo gluten challenge is the gold standard for the assessment of celiac toxicity; however, jejunal biopsy is a relatively invasive procedure, thus, other methods have been investigated. Direct infusion of the rectum with gluten has been shown to result in an increase in mucosal intraepithelial lymphocytes, occurring only in celiac patients. This method has been used to study the celiac toxicity of gliadin subfractions. The in vitro technique of small intestinal biopsy organ culture is also a useful tool and appears to give the same results as in vivo challenge. The importance of tiny amounts of gliadin in the diet, such as that which occurs in wheat starch, has been studied by in vivo challenge; this technique has clarified the position of oats in the gluten-free diet. Several studies suggest that this cereal may be included in the diet of most adult celiac patients. Studies of the transport of gliadin across the enterocyte following ingestion or challenge suggest that gliadin may be metabolized by a different pathway in celiac disease. This could result in an abnormal presentation to the immune system, triggering a pathogenic rather than a tolerogenic response.

Measurements of the jejunal unstirred layer in normal subjects and patients with celiac disease

1996 ◽  
Vol 270 (3) ◽  
pp. G487-G491 ◽  
Author(s):  
A. Strocchi ◽  
G. Corazza ◽  
J. Furne ◽  
C. Fine ◽  
A. Di Sario ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Layer Thickness ◽  
Villous Atrophy ◽  
Normal Subjects ◽  
Unstirred Layer ◽  
Maximal Velocity ◽  
Normal Rat ◽  
Kinetics Of

Normal intestinal absorption of nutrients requires efficient luminal mixing to deliver solute to the brush border. Lacking such mixing, the buildup of thick unstirred layers over the mucosa markedly retards absorption of rapidly transported compounds. Using a technique based on the kinetics of maltose hydrolysis, we measured the unstirred layer thickness of the jejunum of normal subjects and patients with celiac disease, as well as that of the normal rat. The jejunum of humans and rats was perfused with varying maltose concentrations, and the apparent Michaelis constant (Km) and maximal velocity (Vmax) of maltose hydrolysis were determined from double-reciprocal plots. The true Km of intestinal maltase was determined on mucosal biopsies. Unstirred layer thickness was calculated from the in vivo Vmax and apparent Km and the in vitro Km of maltase. The average unstirred layer thickness of 11 celiac patients (170 micron) was seven times greater than that of 3 controls (25 micron). The unstirred layer of each celiac exceeded that of the controls. A variety of factors could account for the less efficient luminal stirring observed in celiacs. Although speculative, villous contractility could be an important stirring mechanism that would be absent in celiacs with villous atrophy. This speculation was supported by the finding of a relatively thick unstirred layer (mean: 106 micron) in rats, an animal that lacks villous contractility. Because any increase in unstirred layer slows transport of rapidly absorbed compounds, poor stirring appears to represent a previously unrecognized defect that could contribute to malabsorption in celiac disease and, perhaps, in other intestinal disorders.

scholarly journals Interleukin-15 May Be Responsible for Early Activation of Intestinal Intraepithelial Lymphocytes after Oral Infection with Listeria monocytogenes in Rats

1998 ◽  
Vol 66 (12) ◽  
pp. 5677-5683 ◽  
Author(s):  
Kenji Hirose ◽  
Hirohiko Suzuki ◽  
Hitoshi Nishimura ◽  
Akio Mitani ◽  
Junji Washizu ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Cell Receptor ◽  
Intraepithelial Lymphocytes ◽  
Oral Infection ◽  
Epithelial Cell Line ◽  
Intestinal Intraepithelial Lymphocytes ◽  
Interleukin 15 ◽  
Ifn Γ

ABSTRACT Exogenous interleukin-15 (IL-15) stimulates intestinal intraepithelial lymphocytes (i-IEL) from mice to proliferate and produce gamma interferon (IFN-γ) in vitro. To determine whether endogenous IL-15 is involved in activation of i-IEL during intestinal infection, we examined IL-15 synthesis by intestinal epithelial cells (i-EC) after infection with Listeria monocytogenes in rats. In in vitro experiments, invasion of L. monocytogenes into IEC-6 cells, a rat small intestine epithelial cell line, evidently induced IL-15 mRNA expression coincident with nuclear factor κB (NF-κB) activation, which is essential for IL-15 gene expression. IL-15 synthesis was detected in rat i-EC on day 1 after an oral inoculation of L. monocytogenes in vivo. The numbers of T-cell receptor (TCR) γδ+ T cells, NKR.P1+cells, and CD3+ CD8+ αα cells in i-IEL were significantly increased on day 1 after oral infection. The i-IEL from infected rats produced larger amounts of IFN-γ upon stimulation with immobilized anti-TCR γδ or anti-NKR.P1 monoclonal antibodies. These results suggest that IL-15 produced by i-EC may stimulate significant fractions of i-IEL to produce IFN-γ at an early phase of oral infection with L. monocytogenes.

scholarly journals Construction and Clinical Verification of Cytoplasm Protein GFAP as Circulating Tumor Cell Separation target for Pediatric Neuroepithelial Tumors

2020 ◽  
Author(s):  
Yang Zhao ◽  
Feng Jiang ◽  
Qinhua Wang ◽  
Baocheng Wang ◽  
Yipeng Han ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Detection System ◽  
Invasive Procedure ◽  
Circulating Tumor Cell ◽  
Immunofluorescence Assay ◽  
Good Choice ◽  
Neuroepithelial Tumors ◽  
Prediction Function

Abstract BACKGROUND: Pediatric Neuroepithelial Tumors (NT) are one of the most prevalent diseases among children. Developing a highly efficient cerebrospinal fluid (CSF) detection system with diagnosis and prediction function is very important. Circulating tumor cell (CTC) in CSF is a good choice. In contrast to the past use of epithelial EpCAM as CTC separation target, an cytoplasm protein of GFAP antibody was first selected to construct highly-sensitive immunomagnetic liposomes (IMLs). The validation and efficiency of this system in capturing CTCs for NT were measured both in vitro and in vivo. The associations between the numbers of CTCs in patients with their clinical characteristics were further analyzed. RESULTS: Our data show that CTCs can be successfully isolated from CSF and blood samples from 29 children with NT. The numbers of CTCs in CSF were significantly higher than those in blood. The level of CTCs in CSF was related to the type and location of the tumor rather than its stage. Genetic testing in GFAP CTC-DNA by sanger sequencing, q-PCR and NGS methods indicated that the isolated CTCs (GFAP+/EGFR+) are the related tumor cell. For example, the high expression of NPR3 gene in CSF CTC was consistant with tumor tissue. CONCLUSIONS: GFAP-IML isolation of CTCs, combined with an EGFR immunofluorescence assay of antitumor marker, can serve as a brand-new method for the identification of CTCs for brain tumors. Via lumbar puncture, a minimally invasive procedure, this technique can be clinically significant in diagnosis and efficacy assessments of pediatric NT.

scholarly journals Tools Used to Measure the Physical State of Women with Celiac Disease: A Review with a Systematic Approach

2020 ◽  
Vol 17 (2) ◽  
pp. 539
Author(s):  
Alejandro Martínez-Rodríguez ◽  
Daniela Alejandra Loaiza-Martínez ◽  
Javier Sánchez-Sánchez ◽  
Pablo J. Marcos-Pardo ◽  
Soledad Prats ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Selection Process ◽  
Immunoglobulin A ◽  
Clinical History ◽  
Diagnostic Tools ◽  
Intestinal Biopsy ◽  
Specific Protocol ◽  
Small Intestinal Biopsy ◽  
Immunological Disorder ◽  
Meta Analyses

Celiac disease (CD) is an immunological disorder that mainly affects the small intestine, generating an inflammatory process in response to the presence of gluten (a protein). Autoimmune diseases are part of a group of diseases that are difficult to diagnose without a specific protocol or consensus to detect them due to the number of symptoms and diseases with which it has a relationship. Therefore, the aim of this review was to analyze the diagnostic tools of CD used in middle-aged women, to compare the use and effectiveness of the different tools, and to propose a strategy for the use of the tools based on the results found in the literature. The present research followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline. The search was conducted in the following databases: Scielo, PubMed, Web of Science, and Worldwide Science org. In the initial literature search, 2004 titles and relevant abstracts were found. Among them, 687 were duplicates, leaving 1130 articles. Based on the inclusion criteria, only 41 articles passed the selection process; 4 main types of analyses appear in the studies: blood tests, questionnaires, clinical history, and biopsy. It can be said that none of the analyses have a 100% reliability since most of them can present false negatives; therefore, the best way to diagnose celiac disease up to now is through a combination of different tests (Immunoglobulin A and small intestinal biopsy).

The zebrafish brain in research and teaching: a simple in vivo and in vitro model for the study of spontaneous neural activity

2011 ◽  
Vol 35 (2) ◽  
pp. 188-196 ◽  
Author(s):  
R. Vargas ◽  
I. þ. Jóhannesdóttir ◽  
B. Sigurgeirsson ◽  
H. þorsteinsson ◽  
K. Æ. Karlsson
Keyword(s):  
Neural Development ◽  
Neural Circuits ◽  
Brain Slices ◽  
Functional Roles ◽  
Zebrafish Brain ◽  
Vitro Model ◽  
Immunohistochemical Techniques ◽  
Neuronal Groups

Recently, the zebrafish ( Danio rerio ) has been established as a key animal model in neuroscience. Behavioral, genetic, and immunohistochemical techniques have been used to describe the connectivity of diverse neural circuits. However, few studies have used zebrafish to understand the function of cerebral structures or to study neural circuits. Information about the techniques used to obtain a workable preparation is not readily available. Here, we describe a complete protocol for obtaining in vitro and in vivo zebrafish brain preparations. In addition, we performed extracellular recordings in the whole brain, brain slices, and immobilized nonanesthetized larval zebrafish to evaluate the viability of the tissue. Each type of preparation can be used to detect spontaneous activity, to determine patterns of activity in specific brain areas with unknown functions, or to assess the functional roles of different neuronal groups during brain development in zebrafish. The technique described offers a guide that will provide innovative and broad opportunities to beginner students and researchers who are interested in the functional analysis of neuronal activity, plasticity, and neural development in the zebrafish brain.

scholarly journals The Broad Spectrum of Celiac Disease and Gluten Sensitive Enteropathy

2016 ◽  
Vol 89 (3) ◽  
pp. 335-342 ◽  
Author(s):  
Oana Mocan ◽  
Dan L. Dumitrașcu
Keyword(s):  
Celiac Disease ◽  
Viral Infections ◽  
Villous Atrophy ◽  
Intraepithelial Lymphocytes ◽  
Intestinal Biopsy ◽  
Gluten Free ◽  
Serological Tests ◽  
Genetic Transmission ◽  
Intestinal Involvement ◽  
Gluten Sensitive Enteropathy

The celiac disease is an immune chronic condition with genetic transmission, caused by the intolerance to gluten. Gluten is a protein from cereals containing the following soluble proteins: gliadine, which is the most toxic, and the prolamins. The average prevalence is about 1% in USA and Europe, but high in Africa: 5.6% in West Sahara. In the pathogenesis several factors are involved: gluten as external trigger, genetic predisposition (HLA, MYO9B), viral infections, abnormal immune reaction to gluten. Severity is correlated with the number of intraepithelial lymphocytes, cryptic hyperplasia and villous atrophy, as well as with the length of intestinal involvement. The severity is assessed according to the Marsh–Oberhuber staging. Diagnostic criteria are: positive serological tests, intestinal biopsy, the reversal after gluten free diet (GFD). Beside refractory forms, new conditions have been described, like the non celiac gluten intolerance. In a time when more and more people adhere to GFD for nonscientific reasons, practitioners should be updated with the progress in celiac disease knowledge.       

scholarly journals The Diverse Potential of Gluten from Different Durum Wheat Varieties in Triggering Celiac Disease: A Multilevel In Vitro, Ex Vivo and In Vivo Approach

Nutrients ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 3566
Author(s):  
Federica Gaiani ◽  
Sara Graziano ◽  
Fatma Boukid ◽  
Barbara Prandi ◽  
Lorena Bottarelli ◽  
...  
Keyword(s):  
Immune Response ◽  
Celiac Disease ◽  
Durum Wheat ◽  
Ex Vivo ◽  
Diet Group ◽  
Control Group ◽  
Wheat Varieties ◽  
Before And After

The reasons behind the increasing prevalence of celiac disease (CD) worldwide are still not fully understood. This study adopted a multilevel approach (in vitro, ex vivo, in vivo) to assess the potential of gluten from different wheat varieties in triggering CD. Peptides triggering CD were identified and quantified in mixtures generated from simulated gastrointestinal digestion of wheat varieties (n = 82). Multivariate statistics enabled the discrimination of varieties generating low impact on CD (e.g., Saragolla) and high impact (e.g., Cappelli). Enrolled subjects (n = 46) were: 19 healthy subjects included in the control group; 27 celiac patients enrolled for the in vivo phase. Celiacs were divided into a gluten-free diet group (CD-GFD), and a GFD with Saragolla-based pasta group (CD-Sar). The diet was followed for 3 months. Data were compared between CD-Sar and CD-GFD before and after the experimental diet, demonstrating a limited ability of Saragolla to trigger immunity, although not comparable to a GFD. Ex vivo studies showed that Saragolla and Cappelli activated immune responses, although with great variability among patients. The diverse potential of durum wheat varieties in triggering CD immune response was demonstrated. Saragolla is not indicated for celiacs, yet it has a limited potential to trigger adverse immune response.

Sign in / Sign up

Export Citation Format

Share Document