scholarly journals Pro-Inflammatory Effect of Gliadins and Glutenins Extracted from Different Wheat Cultivars on an In Vitro 3D Intestinal Epithelium Model

2020 ◽  
Vol 22 (1) ◽  
pp. 172
Author(s):  
Francesca Truzzi ◽  
Camilla Tibaldi ◽  
Anne Whittaker ◽  
Silvia Dilloo ◽  
Enzo Spisni ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Innate Immune Responses ◽  
Wheat Proteins ◽  
Monocyte Migration ◽  
Junction Protein ◽  
Modern Wheat ◽  
In Vitro Effects ◽  
Landrace Wheat ◽  
Cluster Of Differentiation

There is a need to assess the relationship between improved rheological properties and the immunogenic potential of wheat proteins. The present study aimed to investigate the in vitro effects of total protein extracts from three modern and two landrace Triticum aestivum commercial flour mixes, with significant differences in gluten strength (GS), on cell lines. Cytotoxicity and innate immune responses induced by wheat proteins were investigated using Caco-2 monocultures, two dimensional (2D) Caco-2/U937 co-cultures, and three dimensional (3D) co-cultures simulating the intestinal mucosa with Caco-2 epithelial cells situated above an extra-cellular matrix containing U937 monocytes and L929 fibroblasts. Modern wheat proteins, with increased GS, significantly reduced Caco-2 cell proliferation and vitality in monoculture and 2D co-cultures than landrace proteins. Modern wheat proteins also augmented Caco-2 monolayer disruption and tight junction protein, occludin, redistribution in 3D co-cultures. Release of interleukin-8 into the cell medium and increased U937 monocyte migration in both 2D and 3D co-cultures were similarly apparent. Immuno-activation of migrating U937 cells was evidenced from cluster of differentiation 14 (CD14) staining and CD11b-related differentiation into macrophages. The modern wheat proteins, with gluten polymorphism relatedness and increased GS, were shown to be more cytotoxic and immunogenic than the landrace wheat proteins.

scholarly journals In vitro enteroid-derived three-dimensional tissue model of human small intestinal epithelium with innate immune responses

PLoS ONE ◽  
2017 ◽  
Vol 12 (11) ◽  
pp. e0187880 ◽  
Author(s):  
Ying Chen ◽  
Wenda Zhou ◽  
Terrence Roh ◽  
Mary K. Estes ◽  
David L. Kaplan
Keyword(s):  
Immune Responses ◽  
Intestinal Epithelium ◽  
Three Dimensional ◽  
Innate Immune ◽  
Innate Immune Responses ◽  
Tissue Model ◽  
Small Intestinal Epithelium ◽  
Small Intestinal

scholarly journals CD14 is a coreceptor of Toll-like receptors 7 and 9

2010 ◽  
Vol 207 (12) ◽  
pp. 2689-2701 ◽  
Author(s):  
Christoph L. Baumann ◽  
Irene M. Aspalter ◽  
Omar Sharif ◽  
Andreas Pichlmair ◽  
Stephan Blüml ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Innate Immune ◽  
Confocal Imaging ◽  
Acid Uptake ◽  
Innate Immune Responses ◽  
Toll Like Receptors ◽  
Molecular Patterns ◽  
Cluster Of Differentiation

Recognition of pathogens by the innate immune system requires proteins that detect conserved molecular patterns. Nucleic acids are recognized by cytoplasmic sensors as well as by endosomal Toll-like receptors (TLRs). It has become evident that TLRs require additional proteins to be activated by their respective ligands. In this study, we show that CD14 (cluster of differentiation 14) constitutively interacts with the MyD88-dependent TLR7 and TLR9. CD14 was necessary for TLR7- and TLR9-dependent induction of proinflammatory cytokines in vitro and for TLR9-dependent innate immune responses in mice. CD14 associated with TLR9 stimulatory DNA in precipitation experiments and confocal imaging. The absence of CD14 led to reduced nucleic acid uptake in macrophages. Additionally, CD14 played a role in the stimulation of TLRs by viruses. Using various types of vesicular stomatitis virus, we showed that CD14 is dispensable for viral uptake but is required for the triggering of TLR-dependent cytokine responses. These data show that CD14 has a dual role in nucleic acid–mediated TLR activation: it promotes the selective uptake of nucleic acids, and it acts as a coreceptor for endosomal TLR activation.

scholarly journals The Effect of Rutin and Extracts of Uncaria guianensis (Aubl.) J. F. Gmeland on Primary Endometriotic Cells: A 2D and 3D Study

Molecules ◽  
2020 ◽  
Vol 25 (6) ◽  
pp. 1325
Author(s):  
Camila Hernandes ◽  
Renata Nascimento de Oliveira ◽  
Artur Henrique de Souza Santos ◽  
Helena Malvezzi ◽  
Bruna Cestari de Azevedo ◽  
...  
Keyword(s):  
Growth Factor ◽  
Three Dimensional ◽  
Reproductive Age ◽  
Leaf Extracts ◽  
Epidermal Growth ◽  
In Vitro Effects ◽  
Factor Α ◽  
Gynecological Disorder ◽  
2D And 3D

There is increasing interest in the potential of natural compounds to treat diseases, such as endometriosis, a gynecological disorder that affects 10–15% of women of reproductive age, and it is related to severe pelvic pain and infertility. We have evaluated the in vitro effects of rutin and the aqueous bark, roots, and leaf extracts (ABE, ARE, and ALE, respectively) and isolated components of Uncaria guianensis on stromal cells from eutopic endometrium and lesions of patients with endometriosis. Two- and three-dimensional cultures were used to assess the cell death and production of reactive oxygen species (ROS), cytokines and growth factors of cells following exposure to these natural products. The applied treatments did not reduce cellular viability, but ROS production did increase. In addition, significant increases in the levels of interleukin (IL)-15, IL-17A, IL-4, IL-6, tumor necrosis factor-α, and vascular endothelium growth factor were observed when 2D-cells from endometrium of patients with endometriosis were treated with ABE, while exposure to ALE induced significant increases in epidermal growth factor in lesion cells.

Algerian propolis extracts: Chemical composition, bactericidal activity and in vitro effects on gilthead seabream innate immune responses

2017 ◽  
Vol 62 ◽  
pp. 57-67 ◽  
Author(s):  
EL-Khamsa Soltani ◽  
Rebeca Cerezuela ◽  
Noureddine Charef ◽  
Samia Mezaache-Aichour ◽  
Maria Angeles Esteban ◽  
...  
Keyword(s):  
Chemical Composition ◽  
Immune Responses ◽  
Bactericidal Activity ◽  
Innate Immune ◽  
Gilthead Seabream ◽  
Innate Immune Responses ◽  
In Vitro Effects

scholarly journals Comparative Analysis of Cell–Cell Contact Abundance in Ovarian Carcinoma Cells Cultured in Two- and Three-Dimensional In Vitro Models

Biology ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 446
Author(s):  
Olga M. Kutova ◽  
Ludmila M. Sencha ◽  
Anton D. Pospelov ◽  
Olga E. Dobrynina ◽  
Anna A. Brilkina ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Connexin 43 ◽  
Three Dimensional ◽  
3D Models ◽  
In Vitro Models ◽  
Cell Contact ◽  
Tumor Resistance ◽  
Ovarian Adenocarcinoma ◽  
Junction Protein

Tumor resistance to therapy is associated with the 3D organization and peculiarities of the tumor microenvironment, of which intercellular adhesion is a key participant. In this work, the abundance of contact proteins was compared in SKOV-3 and SKOV-3.ip human ovarian adenocarcinoma cell lines, cultivated in monolayers, tumor spheroids and collagen hydrogels. Three-dimensional models were characterized by extremely low expression of basic molecules of adherens junctions E-cadherin and demonstrated a simultaneous decrease in desmosomal protein desmoglein-2, gap junction protein connexin-43 and tight junction proteins occludin and ZO-1. The reduction in the level of contact proteins was most pronounced in collagen hydrogel, accompanied by significantly increased resistance to treatment with doxorubicin and targeted anticancer toxin DARPin-LoPE. Thus, we suggest that 3D models of ovarian cancer, especially matrix-based models, tend to recapitulate tumor microenvironment and treatment responsiveness to a greater extent than monolayer culture, so they can be used as a highly relevant platform for drug efficiency evaluation.

scholarly journals Meprin A impairs epithelial barrier function, enhances monocyte migration, and cleaves the tight junction protein occludin

2013 ◽  
Vol 305 (5) ◽  
pp. F714-F726 ◽  
Author(s):  
Jialing Bao ◽  
Renee E. Yura ◽  
Gail L. Matters ◽  
S. Gaylen Bradley ◽  
Pan Shi ◽  
...  
Keyword(s):  
Tight Junction ◽  
Barrier Function ◽  
Fluorescein Isothiocyanate ◽  
Tight Junction Protein ◽  
Sodium Fluorescein ◽  
Monocyte Migration ◽  
Junction Protein ◽  
Epithelial Barriers

Meprin metalloproteases are highly expressed at the luminal interface of the intestine and kidney and in certain leukocytes. Meprins cleave a variety of substrates in vitro, including extracellular matrix proteins, adherens junction proteins, and cytokines, and have been implicated in a number of inflammatory diseases. The linkage between results in vitro and pathogenesis, however, has not been elucidated. The present study aimed to determine whether meprins are determinative factors in disrupting the barrier function of the epithelium. Active meprin A or meprin B applied to Madin-Darby canine kidney (MDCK) cell monolayers increased permeability to fluorescein isothiocyanate-dextran and disrupted immunostaining of the tight junction protein occludin but not claudin-4. Meprin A, but not meprin B, cleaved occludin in MDCK monolayers. Experiments with recombinant occludin demonstrated that meprin A cleaves the protein between Gly100 and Ser101 on the first extracellular loop. In vivo experiments demonstrated that meprin A infused into the mouse bladder increased the epithelium permeability to sodium fluorescein. Furthermore, monocytes from meprin knockout mice on a C57BL/6 background were less able to migrate through an MDCK monolayer than monocytes from their wild-type counterparts. These results demonstrate the capability of meprin A to disrupt epithelial barriers and implicate occludin as one of the important targets of meprin A that may modulate inflammation.

Ultrastructural Investigations of the Contractile Filaments of Amoebae

1974 ◽  
Vol 32 ◽  
pp. 188-189
Author(s):  
P.L. Moore
Keyword(s):  
Cytoplasmic Streaming ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Amoeboid Movement ◽  
Different Types ◽  
Chemical Conditions ◽  
Complete Interpretation

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.

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