HT-29 cells: a new substrate for rotavirus growth

Superti, Fabiana; Tinari, Antonella; Baldassarri, Lucilla; Donelli, Gianfranco

doi:10.1007/bf01319239

Cited by 22 publications

(18 citation statements)

References 42 publications

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“…Many integrin-using and integrin-independent rotaviruses replicate to high titer in both Caco-2 and HT-29 cells (43,66). It is noteworthy that ␣V␤3 is absent from HT-29 cells, indicating that ␣V␤3 is not necessary for RRV entry.…”

Section: Discussionmentioning

confidence: 96%

Rotavirus Replication in Intestinal Cells Differentially Regulates Integrin Expression by a Phosphatidylinositol 3-Kinase-Dependent Pathway, Resulting in Increased Cell Adhesion and Virus Yield

Halász

Holloway

Turner

et al. 2008

J Virol

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Changes in the interactions between intestinal cells and their surrounding environment during virus infection have not been well documented. The growth and survival of intestinal epithelial cells, the main targets of rotavirus infection, are largely dependent on the interaction of cell surface integrins with the extracellular matrix. In this study, we detected alterations in cellular integrin expression following rotavirus infection, identified the signaling components required, and analyzed the subsequent effects on cell binding to the matrix component collagen. After rotavirus infection of intestinal cells, expression of ␣2␤1 and ␤2 integrins was up-regulated, whereas that of ␣V␤3, ␣V␤5, and ␣5␤1 integrins, if present, was down-regulated. This differential regulation of integrins was reflected at the transcriptional level. It was unrelated to the use of integrins as rotavirus receptors, as both integrin-using and integrin-independent viruses induced integrin regulation. Using pharmacological agents that inhibit kinase activity, integrin regulation was shown to be dependent on phosphatidylinositol 3-kinase (PI3K) but independent of the activities of the mitogen-activated protein kinases p38 and ERK1/2, and cyclooxygenase-2. Replication-dependent activation of the PI3K/Akt pathway was observed following infection of intestinal and nonintestinal cell lines. Rotavirus activation of PI3K was important for regulation of ␣2␤1 expression. Blockade of integrin regulation by PI3K inhibition led to decreased adherence of infected intestinal cells to collagen and a concomitant decrease in virus titer. These findings indicate that rotavirus-induced PI3K activation causes regulation of integrin expression in intestinal cells, leading to prolonged adherence of infected cells to collagen and increased virus production.Rotavirus is a major cause of dehydrating gastroenteritis in infant humans and animals throughout the world. Differentiated epithelial cells on villi in the small intestine (enterocytes) are the main targets of infection, leading to cell death, a reduction in villus epithelium area, loss of absorptive capacity, and osmotic dysregulation (3,7,48,53). Enterocytes are attached to the basement membrane through interactions between the basement matrix and cell-surface adhesion molecules, mainly members of the integrin family (4,6,49,64). Increased enterocyte loss is a feature of rotavirus disease, suggesting that reduced enterocyte adhesion occurs (9). However, the molecules involved in changes in attachment of enterocytes to the matrix during rotavirus infection and how this process relates to viral replication and pathogenesis have not been analyzed.Integrins provide a means for cells to respond to their environment through intracellular signaling networks controlling crucial cellular processes such as adhesion, proliferation, migration, differentiation, and survival (1,20,38,55,69). These ␣␤ heterodimeric glycoproteins have been identified as cellular receptors or entry cofactors for many viruses, including rotavi...

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Section: Discussionmentioning

confidence: 96%

Rotavirus Replication in Intestinal Cells Differentially Regulates Integrin Expression by a Phosphatidylinositol 3-Kinase-Dependent Pathway, Resulting in Increased Cell Adhesion and Virus Yield

Halász

Holloway

Turner

et al. 2008

J Virol

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“…We initially tested the antiviral activity of these proteins in HT-29 cells which, in spite of their colonic derivation, express the differentiation phenotype of mature enterocytes [30,31] and are highly susceptible to rotavirus infection [30], thus representing a suitable in vitro model for investigating the infectious pathways of this pathogen. In this cell system, viral c.p.e.…”

Section: Discussionmentioning

confidence: 99%

Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29

Superti

Ammendolia

Valenti

et al. 1997

Medical Microbiology and Immunology

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169

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Different milk proteins were analyzed for their inhibitory effect on either rotavirus-mediated agglutination of human erythrocytes or rotavirus infection of the human enterocyte-like cell line HT-29. Proteins investigated were alpha-lactalbumin, beta-lactoglobulin, apo-lactoferrin, and Fe(3+)-lactoferrin, and their antiviral action was compared with the activity of mucin, a milk glycoprotein known to affect rotavirus infection. Results obtained demonstrated that beta-lactoglobulin, apo- and Fe(3+)-lactoferrin are able to inhibit the replication of rotavirus in a dose-dependent manner, apo-lactoferrin being the most active. It was shown that apo-lactoferrin hinders virus attachment to cell receptors since it is able to bind the viral particles and to prevent both rotavirus haemagglutination and viral binding to susceptible cells. Moreover, this protein markedly inhibited rotavirus antigen synthesis and yield in HT-29 cells when added during the viral adsorption step or when it was present in the first hours of infection, suggesting that this protein interferes with the early phases of rotavirus infection.

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“…Likewise, HEp2 cells are epithelial cells that are highly permissive for picornaviruses but relatively resistant to rotavirus infection. HT29 human intestinal epithelial cells have been previously reported to be useful for the isolation of human rotaviruses ( 14).…”

Section: Methodsmentioning

confidence: 99%

Liposome-mediated transfection of intact viral particles reveals that plasma membrane penetration determines permissivity of tissue culture cells to rotavirus.

et al. 1992

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Rotaviruses are an important cause of gastroenteritis in human infants. In vivo, rotavirus displays striking cell tropism with viral replication generally restricted to the villus tip enterocytes of the small intestine. We studied a panel of cell lines that vary significantly in their permissivity to rotavirus infection. L cells and HEp2 cells were relatively resistant to rotavirus infection compared with permissive MalO4 cells and HT29 cells. RNA transcription among the cell lines was proportional to antigen synthesis making a translational or posttranslational block an unlikely source of observed differences in susceptibility. All of the cell lines bound and internalized radiolabeled virus equally well, as measured by escape from surface protease treatment. Analysis of the escape of cell bound virus from neutralizing monoclonal antibody revealed that rotavirus did not immediately enter an eclipse phase in nonpermissive cells, but was internalized in an infectious form for several hours, possibly sequestered within endocytic vacuoles. L cells and HEp2 cells were as permissive as MalO4 and HT29 cells when rotavirus infection was mediated by transfection of single-or doubleshelled rotavirus particles with cationic liposomes (LipofectinTM). Rotavirus cell tropism in tissue culture cells is determined by the ability of infecting virions to traverse the plasma membrane of the cells into the cytoplasmic compartment. (J.

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HT-29 cells: a new substrate for rotavirus growth

Cited by 22 publications

References 42 publications

Rotavirus Replication in Intestinal Cells Differentially Regulates Integrin Expression by a Phosphatidylinositol 3-Kinase-Dependent Pathway, Resulting in Increased Cell Adhesion and Virus Yield

Rotavirus Replication in Intestinal Cells Differentially Regulates Integrin Expression by a Phosphatidylinositol 3-Kinase-Dependent Pathway, Resulting in Increased Cell Adhesion and Virus Yield

Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29

Liposome-mediated transfection of intact viral particles reveals that plasma membrane penetration determines permissivity of tissue culture cells to rotavirus.

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