Structural Analysis of a Human Intestinal Epithelial Cell Line

Madara, James L.; Stafford, J; Dharmsathaphorn, Kiertisin; Carlson, Susan

doi:10.1016/s0016-5085(87)91069-9

Cited by 186 publications

(135 citation statements)

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“…Of special interest in this context are polar compounds that may be subject to significant diffusion restriction by the formation of a 'tight' epithelial barrier (Madara et al, 1987). Thus inulin possesses an extremely low permeability in comparison to vinblastine, whose transepithelial permeability is likely to be predominantly transcellular in such highly resistive layers.…”

Section: Immunoprecipation Of P-glycoproteinmentioning

confidence: 99%

“…Calcium antagonists (e.g., verapamil), calmodulin inhibitors (e.g., trifluoperazine) and other agents (e.g., reserpine) inhibit the active drug efflux and restore drug sensitivity in multidrug resistant cells (Tsuruo, 1983;Kessel & Wilberding, 1984;Kessel & Wilberding, 1985;Bellamy et al, 1988;Twentyman et al, 1986). (Madara et al, 1987;Chan et al, 1989 Dharmsathaphorn et al (1984) and Simmons (1990). Cells were seeded at high density (1 x 106 cells ml-', 3 ml well) into rat-tail collagen coated filtercups, Transwell 24.5 mm diam.…”

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confidence: 99%

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Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Hunter

Hirst²,

Simmons³

1991

Br J Cancer

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Summary P-glycoprotein expression was demonstrated in two human intestinal adenocarcinoma cell-lines ileocaecal and T84, colonic) by immunoprecipitation of a 170-180 kDa protein with monoclonal antibody JSB-1. Both HCT-8 and T84 formed functional epithelial cell layers of high transepithelial electrical resistance (>700 Q cm2) when grown on permeable matrices. These epithelial layers demonstrated vectorial secretion (net vinblastine fluxes in the basal-to-apical direction of 0.135 and 0.452 pmol h-' cm-2 in HCT-8 and T84 cell layers, respectively, from bathing solutions containing 10 nM vinblastine). These vectorial vinblastine secretions were sensitive to inhibition by verapamil. Passive transepithelial vinblastine permeation was limited by the presence of intercellular (tight) junctions, as demonstrated by the high transepithelial electrical resistance, and verapamil increased this passive vinblastine permeation concomitant with a reduction in the electrical resistance. Cellular vinblastine loading was significantly greater from the basal side, and this was also susceptible to inhibition by basal verapamil. The demonstration of vectorial transport of vinblastine in human intestinal colonic adenocarcinoma cell layers is direct evidence in favour of the hypothesis that the function of mdrl in epithelia from the gastrointestinal tract is to promote detoxification by a process of epithelial secretion. This study also highlights that cellular vinblastine accumulation depends not only upon P-glycoprotein function, but also upon differential apparent membrane permeabilities and the presence of intercellular (tight) junctions that may restrict drug permeation and cellular accumulation to apical or basal membrane domains.

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Section: Immunoprecipation Of P-glycoproteinmentioning

confidence: 99%

mentioning

confidence: 99%

Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Hunter

Hirst²,

Simmons³

1991

Br J Cancer

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“…We have utilised T84 cells to model the epithelial barrier. 29 T84 cells have structural similarities to intestinal crypt cells 39 and increase their tight junction strand number with time in monolayer culture that correlates with increased resistance. 40 Since intestinal crypt cells are believed to be leaky relative to surface cells [41][42][43] it is unclear how the developing monolayer culture model relates to the differentiation of cells in vivo, but it may be representative of the more differentiated epithelium in relation to tight junction assembly.…”

mentioning

confidence: 99%

Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells

Prasad

Mingrino

Kaukinen

et al. 2005

Laboratory Investigation

419

384

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Claudin proteins comprise a recently described family of tight junction proteins that differentially regulate paracellular permeability. Since other tight junction proteins show alterations in distribution or expression in inflammatory bowel disease (IBD) we assessed expression of claudins (CL) 2, 3 and 4 in IBD. CL 2 was strongly expressed along the inflamed crypt epithelium, whilst absent or barely detectable in normal colon. In contrast, CL 3 and 4 were present throughout normal colonic epithelium and were reduced or redistributed in the diseased surface epithelium. In a T84-cell culture model of the gut barrier, paracellular permeability decreased with time after plating and correlated with a marked decrease in the expression of CL 2. Addition of IFNc/TNFa led to further decreases in CL 2 and 3, the redistrbution of CL 4 and a marked increase in paracellular permeability. Conversely, IL-13 dramatically increased CL 2, with little effect on CL 3 or 4, but also resulted in increased paracellular permeability. Expression of CL 2 did not correlate with proliferation or junctional reorganisation after calcium ion depletion. Re-expression of CL 2 in response to IL-13 was inhibited by phophatidylinositol 3 kinase inhibitor, LY294002, which also restored the ion permeability to previous levels. CL 2 expression could be stimulated in the absence of IL-13 by activation of phospho-Akt in the phophatidylinositol 3 kinase pathway. These results suggest that INFc/TNFa and IL-13 have differential effects on CL 2, 3 and 4 in tight junctions, which may lead to increased permeability via different mechanisms.

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“…Such monolayers are composed of columnar epithelial cells with features similar to those of natural crypt epithelia (Madara and Dharmsathaphorn, 1985;Madara et al, 1987). These cells join neighbors through apical circumferential tight junctions which maintain structure-function characteristics found in fight junctions of natural epithelia (Madara and Dharmsathaphorn, 1985).…”

mentioning

confidence: 99%

Neutrophil migration across cultured intestinal epithelial monolayers is modulated by epithelial exposure to IFN-gamma in a highly polarized fashion.

Colgan

Parkos

Delp

et al. 1993

The Journal of Cell Biology

158

111

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Abstract. Neutrophil, or polymorphonuclear leukocyte (PMN), migration across intestinal epithelial barriers, such as occurs in many disease states, appears to result in modifications of epithelial barrier and ion transport functions (Nash, S., J. Stafford, and J. L. . Z Clin. Invest. 80:1104-1113; Madara, J. L., C. A. Parkos, S. P. Colgan, R. J. MacLeod, S. Nash, J. B. Matthews, C. Delp, and W. I. Lencer. 1992. J. Clin. Invest. 89:1938-1944). Here we investigate the effects of epithelial exposure to IFN-,), on PMN migration across cultured monolayers of the human intestinal epithelial cell line Tu. Transepithelial migration of PMN was initially assessed in the apicalto-basolateral direction, since previous studies indicate general qualitative similarities between PMN migration in the apical-to-basolateral and in the basolateralto-apical directions. In the apical-to-basolateral direction, epithelial exposure to IFN-.,/rnarkedly upregulated transepithelial migration of PMN in a dose-and timedependent fashion as measured by both electrical and myeloperoxidase assays. This IFN-~/-elicited effect on transmigration was specifically due to a IFN-,/effect on epithelial cells and was not secondary to IFN-,y effects on epithelial tight junction permeability. Moreover, this IFN-.y effect was dependent on epithelial protein synthesis, and involved a pathway in which CDllb/18, but not ICAM-1 or CDlla/18, appeared to play a crucial role in PMN-epithelial adhesion. IFN-~/also substantially modified PMN transepithelial migration in the natural, basolateral-to-apical direction. The IFN-'y effect on naturally directed transmigration was also specifically due to an IFN-3, effect on epithelial cells, showed comparable time and dose dependency to that of oppositely directed migration, was CDllb/18 dependent, and required epithelial protein synthesis. Additionally, however, important qualitative differences existed in how IFN-3, affected transmigration in the two directions. In contrast to apical-to-basolateral directed migration, IFN-,), markedly downregulated transepithelial migration of PMN in the natural direction. This downregulation of PMN migration in the natural direction, however, was not due to failure of PMN to move across filters and into monolayers. Indeed, IFN-3, exposure to epithelia increased the number of PMN which had moved into the basolateral space of the epithelium in naturally directed transmigration. These results represent the first detailed report of influences on PMN transepithelial migration by a cytokine, define conditions under which a qualitative difference in PMN transepithelial migration exists, and suggest that migration of PMN across epithelia in the natural direction may involve multiple steps which can be differentially regulated by cytokines. Specifically, it appears that in naturally directed migration, IFN-~/ may enhance the retention time of the recruited PMN in the paracellular space below tight junctions, and does so, at least in part, by downregulating a PMN-epithelial interactive event req...

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Structural Analysis of a Human Intestinal Epithelial Cell Line

Cited by 186 publications

References 10 publications

Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Epithelial secretion of vinblastine by human intestinal adenocarcinoma cell (HCT-8 and T84) layers expressing P-glycoprotein

Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells

Neutrophil migration across cultured intestinal epithelial monolayers is modulated by epithelial exposure to IFN-gamma in a highly polarized fashion.

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