Early tissue transglutaminase–mediated response underlies K562(S)-cell gliadin-dependent agglutination

Silano, Marco; Vincentini, Olimpia; Luciani, Alessandro; Felli, Cristina; Caserta, Sergio; Esposito, Speranza; Villella, Valeria Rachela; Pettoello-Mantovani, Massimo; Guido, Stefano; Maiuri, Luigi

doi:10.1038/pr.2012.4

Cited by 32 publications

(34 citation statements)

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“…We have previously shown [27] that the agglutination of myelogenic leukemia cells K562(S) by toxic gliadin peptides relies upon the activity of cellular TG2, an enzyme involved in the very precocious events triggered by these peptides at the cell surface of duodenal epithelial cells. Briefly, the Ca 2+ -dependent TG2 activation triggered by p31–43 in K562 cells leads to cytoskeleton rearrangement, which is a key event in cell agglutination.…”

Section: Resultsmentioning

confidence: 99%

“…Human leukemic K562(S) and human colon adenocarcinoma T84 cell lines were obtained from American Type Culture Collection (commercialized in Italy by LGC Standard, Milan) and cultured as previously described [27]. …”

Section: Methodsmentioning

confidence: 99%

“…The K562(S) cell agglutination test was performed as previously described [27]. Briefly, K562(S) cells were harvested by centrifugation and washed twice with Ca 2+ and Mg 2+ -free PBS (GIBCO, Carlsbad, CA, USA) and resuspended at a concentration of 10 8 cells/mL in the same PBS.…”

Section: Methodsmentioning

confidence: 99%

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Diversity of oat varieties in eliciting the early inflammatory events in celiac disease

et al. 2013

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Purpose Celiac disease (CD) is an autoimmune enteropathy, triggered by dietary gluten. The only treatment is a strict gluten-free diet. Oats are included in the list of gluten-free ingredients by European Regulation, but the safety of oats in CD is still a matter of debate. The present study examined the capability of different oat cultivars of activating the gliadin-induced transglutaminase-2 (TG2)-dependent events in some in vitro models of CD. In addition, we compared this capability with the electrophoresis pattern of peptic–tryptic digests of the proteins of the oat cultivars.MethodsK562(S) cells agglutination, transepithelial electrical resistance of T84-cell monolayers, intracellular levels of TG2 and phosphorylated form of protein 42–44 in T84 cells were the early gliadin-dependent events studied.ResultsThe results showed that the Nave oat cultivar elicited these events, whereas Irina and Potenza varieties did not. The ability of a cultivar to activate the above-described events was associated with the electrophoretic pattern of oat proteins and their reactivity to anti-gliadin antibodies.ConclusionWe found significant differences among oat cultivars in eliciting the TG2-mediated events of CD inflammation. Therefore, the safety of an oat cultivar in CD might be screened in vitro by means of biochemical and biological assays, before starting a clinical trial to definitely assess its safety.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Diversity of oat varieties in eliciting the early inflammatory events in celiac disease

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“…[32][33][34] The microscope (Zeiss Axiovert 200, 103 and 203 objectives) is equipped with a high sensitivity cooled CCD camera (Hamamtsu Orca AG) and motorized stage, focus (Marzhauser) and filter wheels (Prior) for automated 6-D Time Lapse imaging. The stage repositioning error (repeatability: < 1 lm, resolution: 0.01 lm, accuracy: 6 3 lm, data from the manufacturer), is below the optical resolution of our experiments, due to the pixel size (0.5-1.0 lm, depending on the optic used), and we can consider the overall planar repositioning error to be around 1 lm.…”

Section: Time Lapse Microscopymentioning

confidence: 99%

A methodology to study chemotaxis in 3‐D collagen gels

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We present here an innovative experimental methodology for the quantitative investigation of chemotaxis in vitro by live imaging of cell movement in a reconstituted three-dimensional collagen gel. A well-defined chemoattractant gradient is generated by means of a novel direct viewing chamber having two compartments (separated by a membrane), one containing the chemoattractant solution, the other the cell-seeded collagen gel matrix. Cell migration is observed by means of a time-lapse motorized video-microscopy workstation equipped with an incubating system and quantified by image analysis techniques. Experimental results on three different cell lines (Jurkat, fibroblasts, and lymphocytes) are presented for the isotropic control case (no chemoattractant) and in presence of a concentration gradient. Cell motility data are in line with the concentration profile, both theoretically calculated from Fick’s law and experimentally measured by epifluorescence microscopy. In particular, a transient peak in cell response was found, possibly due to cell membrane receptor saturation

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“…14,29 The effect of TG-2 on cell adhesion has been reported in NIH mouse 3T3 fibroblasts, 19 human umbilical endothelial cells, 29 human corneal epithelial cells, 24 astrocytes, 30 and a myelogenous leukemia cell line. 31 Despite the possible multiplicity of extracellular and intrinsic factors that exist in different tissues, some aspects of the TG-2-paxillin relationship 24 may be evolutionarily conserved. If so, this commentary addresses a fundamental issue, which will likely impact adhesion and cellular behavior in biology.…”

Section: Transglutaminase-2 Affects Adhesion In Diverse Cell Typesmentioning

confidence: 99%

Transglutaminase-2 in cell adhesion

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Tong

2013

Cell Adhesion & Migration

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Early tissue transglutaminase–mediated response underlies K562(S)-cell gliadin-dependent agglutination

Cited by 32 publications

References 35 publications

Diversity of oat varieties in eliciting the early inflammatory events in celiac disease

Diversity of oat varieties in eliciting the early inflammatory events in celiac disease

A methodology to study chemotaxis in 3‐D collagen gels

Transglutaminase-2 in cell adhesion

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