Celiac Disease

1st Edition, Volume 179 - August 1, 2023
Editors: Ainara Castellanos-Rubio, Lorenzo Galluzzi
Language: English
Hardback ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 2 0 2 - 9
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 1 9 2 0 3 - 6

Celiac Disease, Volume 179 in the Methods in Cell Biology series is composed of extensive protocols about novel and widely used techniques for celiac disease research. Topics… Read more

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Celiac Disease, Volume 179 in the Methods in Cell Biology series is composed of extensive protocols about novel and widely used techniques for celiac disease research. Topics covered in this volume include Pepsin trypsin digested gliadin treatment of intestinal cells in vitro, Ex vivo Gliadin Stimulation of Intestinal Cells, Measuring intestinal permeability in celiac disease ex vivo using Ussing chambers, Intestinal permeability assessment using lactulose and mannitol, In vivo sensitization to gliadin by oral administration, Separation of epithelial and immune cells from intestinal biopsy samples, Isolation and study of dendritic cells and macrophages from human intestinal samples, Gnobiotic mouse models to study gluten sensitivity, and more.

Other sections in this new release include In vitro differentiation of macrophages from peripheral blood cells of celiac patients, Assessment of activated gut-homing CD8+ T cells in blood by flow cytometry during a three-day gluten challenge, Isolation, cryopreservation and thawing of peripheral blood mononuclear cells for downstream applications, and more.

Cover image
Title page
Table of Contents
Series Page
Copyright
Contributors
Chapter 1: Pepsin-trypsin digested gliadin treatment in intestinal cells
Abstract
1: Introduction
2: Cell maintenance
3: PTG treatment
4: RNA and protein extraction and analysis
5: Concluding remarks
6: Notes
Acknowledgments
References
Chapter 2: Ex vivo gliadin stimulation of intestinal cells
Abstract
1: Introduction
2: Reagents
3: Mouse intestinal epithelial cell extraction
4: Gliadin stimulation
5: Concluding remarks
References
Chapter 3: Measuring intestinal permeability in celiac disease ex vivo, using Ussing chambers
Abstract
1: Introduction
2: Protocol
3: Additional notes
Acknowledgments
References
Chapter 4: Intestinal permeability assessment using lactulose and mannitol in celiac disease
Abstract
1: Introduction
2: The lactulose and mannitol (LM) test
3: Sugar analysis by ion chromatography using the technique High-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD)
4: Interpretation of results
5: Concluding remarks
6: Notes
Acknowledgments
References
Chapter 5: In vivo sensitization to gliadin by oral administration
Abstract
1: Introduction
2: Animals
3: Reagents
4: Gliadin sensitization process
5: Validation of the gliadin sensitization process
6: Notes
Acknowledgments
References
Chapter 6: Separation of epithelial and immune cells from biopsy samples
Abstract
1: Introduction
2: Biopsy sample collection and processing
3: Removal of dead cells
4: Leucocyte labeling
5: Magnetic separation
6: Sample storage
7: Labeling for flow cytometry
8: Concluding remarks
9: Notes
Acknowledgments
References
Chapter 7: Study and isolation of human intestinal dendritic cell and macrophage subsets
Abstract
1: Introduction
2: Study of DC and MΦ from intestinal biopsies
3: DC and MΦ isolation from intestinal resections
4: Notes
Acknowledgments
References
Chapter 8: Elucidating the role of microbes in celiac disease through gnotobiotic modeling
Abstract
1: Introduction
2: Animal models of celiac disease
3: Axenic and gnotobiotic models to study celiac disease
4: Generation, maintenance and monitoring of germ-free mice
5: Colonization of germ-free mice
6: Conclusions
References
Chapter 9: In vitro differentiation of macrophages from peripheral blood cells of celiac patients
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
References
Chapter 10: Assessment of activated gut-homing CD8+ T cells in blood by flow cytometry during a 3-day gluten challenge
Abstract
1: Introduction
2: Materials
3: Methods
4: Concluding remarks
5: Notes
Acknowledgments
Competing interests
References
Chapter 11: Isolation and cryopreservation of peripheral blood mononuclear cells
Abstract
1: Introduction
2: Materials (see Note 1)
3: Methods
4: Concluding remarks
Competing interests
References
Chapter 12: Flow cytometric analysis of duodenal intraepithelial lymphocytes (celiac lymphogram): A diagnostic test for celiac disease
Abstract
1: Introduction
2: Materials (see Note 1)
3: Methods
4: Interpretation
5: Notes
6: Concluding Remarks
Acknowledgments
References
Chapter 13: The HLA-DQ8 transgenic mouse: A model to study the immune and cytotoxic responses to wheat gliadin
Abstract
1: Introduction
2: The HLA-DQ8 transgenic mouse model
3: Induction of the enteropathy
4: Induction of the intestinal immune response
5: Cell culture
6: Analyses of selected cytokines (IFN-γ, IL-10, IL-17)
7: Morphology assessment
8: Immunostaining of cryostat sections
9: Concluding remarks
10: Notes
Acknowledgments
References
Chapter 14: Human intestinal organoid models for celiac disease research
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Troubleshooting
6: Concluding remarks
Acknowledgments
Competing interests
References
Chapter 15: Preparation of pepsin trypsin digested gliadin for stimulation experiments
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
References
Chapter 16: A fast, cheap, and easy protocol for celiac disease HLA haplotype typing using buccal swabs
Abstract
1: Introduction
2: Materials
3: Methods
4: HLA haplotype determination
Notes
5: Concluding remarks
References

Ainara Castellanos-Rubio

Ainara Castellanos-Rubio obtained her PhD on Genetics from the University of the Basque Country (Spain) in 2010. During her PhD, she studied gene expression alterations and genetic polymorphisms associated to Celiac Disease. She did a short term research stay in the University of Tampere (Finland) under the supervision of Dr. Marku Makki and Dr. Katri Lindfors where she used three dimensional cell cultures to describe different pathways involved in Celiac Disease development. On 2011 she joined the Laboratory of Dr. Sankar Ghosh in Columbia University (NY, USA) where she carried out her postdoctoral studies. During her postdoctoral training, she studied the implication of long noncoding RNAs (lncRNAs) in the immune and inflammatory response and she discovered and functionally characterized a novel lncRNA involved in the susceptibility to Celiac Disease. Dr. Castellanos-Rubio is an Ikerbasque Associate in the University of the Basque Country, where she leads her own group. She is interested in the involvement of noncoding RNAs in the pathogenesis of autoimmune and inflammatory diseases and studies the influence of disease associated SNPs in the functional disturbance of these RNAs. More recently, she has become interested on the epitranscriptomic alterations involved in different aspects of RNA regulation and her group studies how SNPs and environmental factors can alter these epitranscriptomic signals influencing the inflammatory response that finally evolves in disease development.

Affiliations and expertise

University of the Basque Country and Ikerbasque, Basque Foundation for Science, Leioa, Spain

Lorenzo Galluzzi

Lorenzo Galluzzi is Assistant Professor of Cell Biology in Radiation Oncology at the Department of Radiation Oncology of the Weill Cornell Medical College, Honorary Assistant Professor Adjunct with the Department of Dermatology of the Yale School of Medicine, Honorary Associate Professor with the Faculty of Medicine of the University of Paris, and Faculty Member with the Graduate School of Biomedical Sciences and Biotechnology of the University of Ferrara, the Graduate School of Pharmacological Sciences of the University of Padova, and the Graduate School of Network Oncology and Precision Medicine of the University of Rome “La Sapienza”. Moreover, he is Associate Director of the European Academy for Tumor Immunology and Founding Member of the European Research Institute for Integrated Cellular Pathology. Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. He has published over 450 articles in international peer-reviewed journals and is the Editor-in-Chief of four journals: OncoImmunology (which he co-founded in 2011), International Review of Cell and Molecular Biology, Methods in Cell biology, and Molecular and Cellular Oncology (which he co-founded in 2013). Additionally, he serves as Founding Editor for Microbial Cell and Cell Stress, and Associate Editor for Cell Death and Disease, Pharmacological Research and iScience.

Affiliations and expertise

Assistant Professor of Cell Biology in Radiation Oncology, Department of Radiation Oncology, Weill Cornell Medical College, NY, USA