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Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin.
Am J Physiol. 1999 05; 276(5):G1279-88.AJ

Abstract

The effects of interferon-gamma (IFN-gamma) on tight junctions in T84 human intestinal epithelial cells were investigated. Treatment of T84 cells with IFN-gamma caused a dose- and time-dependent increase in monolayer permeability as assessed by transepithelial electrical resistance measurements. Examination of specific proteins associated with tight junctions by immunoblotting and confocal microscopy revealed changes in the expression levels and localization of some of these proteins after exposure of the cells to IFN-gamma. Specifically, IFN-gamma treatment resulted in an almost total loss of zonula occludens (ZO)-1, whereas the levels of ZO-2 and occludin showed relatively modest decreases compared with untreated cells. Loss of ZO-1 was associated with the altered localization of ZO-2 and occludin. In IFN-gamma-treated cells, ZO-2 and occludin were diffusely distributed, whereas, in control cells, they, along with ZO-1, were predominantly localized to the tight junctions. Analysis of ZO-1 protein and RNA by pulse chase and RT-PCR, respectively, showed an increase in protein turnover, a decrease in protein synthesis, and a reduction in RNA levels following IFN-gamma treatment. In contrast to ZO-1, ZO-2 and occludin did not show any major changes in these parameters. In addition, the organization of actin in the apical and tight junction regions, but not in the mid- or basal regions, of the cells was also perturbed by IFN-gamma treatment of cells. Time-course analysis of IFN-gamma-induced alterations in ZO-1 expression and apical actin perturbation indicated that these two effects were intimately linked and could not be dissociated. These results suggest that IFN-gamma affects barrier function in T84 cells by decreasing the levels of ZO-1 and perturbing apical actin organization, which leads to a disorganization of the tight junction and an increase in paracellular permeability.

Authors+Show Affiliations

Department of Biochemistry, Immunex Corporation, Seattle, Washington 98101, USA. ayouakim@immunex.comNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10330020

Citation

Youakim, A, and M Ahdieh. "Interferon-gamma Decreases Barrier Function in T84 Cells By Reducing ZO-1 Levels and Disrupting Apical Actin." The American Journal of Physiology, vol. 276, no. 5, 1999, pp. G1279-88.
Youakim A, Ahdieh M. Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin. Am J Physiol. 1999;276(5):G1279-88.
Youakim, A., & Ahdieh, M. (1999). Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin. The American Journal of Physiology, 276(5), G1279-88. https://doi.org/10.1152/ajpgi.1999.276.5.G1279
Youakim A, Ahdieh M. Interferon-gamma Decreases Barrier Function in T84 Cells By Reducing ZO-1 Levels and Disrupting Apical Actin. Am J Physiol. 1999;276(5):G1279-88. PubMed PMID: 10330020.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Interferon-gamma decreases barrier function in T84 cells by reducing ZO-1 levels and disrupting apical actin. AU - Youakim,A, AU - Ahdieh,M, PY - 1999/5/18/pubmed PY - 1999/5/18/medline PY - 1999/5/18/entrez SP - G1279 EP - 88 JF - The American journal of physiology JO - Am J Physiol VL - 276 IS - 5 N2 - The effects of interferon-gamma (IFN-gamma) on tight junctions in T84 human intestinal epithelial cells were investigated. Treatment of T84 cells with IFN-gamma caused a dose- and time-dependent increase in monolayer permeability as assessed by transepithelial electrical resistance measurements. Examination of specific proteins associated with tight junctions by immunoblotting and confocal microscopy revealed changes in the expression levels and localization of some of these proteins after exposure of the cells to IFN-gamma. Specifically, IFN-gamma treatment resulted in an almost total loss of zonula occludens (ZO)-1, whereas the levels of ZO-2 and occludin showed relatively modest decreases compared with untreated cells. Loss of ZO-1 was associated with the altered localization of ZO-2 and occludin. In IFN-gamma-treated cells, ZO-2 and occludin were diffusely distributed, whereas, in control cells, they, along with ZO-1, were predominantly localized to the tight junctions. Analysis of ZO-1 protein and RNA by pulse chase and RT-PCR, respectively, showed an increase in protein turnover, a decrease in protein synthesis, and a reduction in RNA levels following IFN-gamma treatment. In contrast to ZO-1, ZO-2 and occludin did not show any major changes in these parameters. In addition, the organization of actin in the apical and tight junction regions, but not in the mid- or basal regions, of the cells was also perturbed by IFN-gamma treatment of cells. Time-course analysis of IFN-gamma-induced alterations in ZO-1 expression and apical actin perturbation indicated that these two effects were intimately linked and could not be dissociated. These results suggest that IFN-gamma affects barrier function in T84 cells by decreasing the levels of ZO-1 and perturbing apical actin organization, which leads to a disorganization of the tight junction and an increase in paracellular permeability. SN - 0002-9513 UR - https://www.unboundmedicine.com/medline/citation/10330020/Interferon_gamma_decreases_barrier_function_in_T84_cells_by_reducing_ZO_1_levels_and_disrupting_apical_actin_ L2 - https://journals.physiology.org/doi/10.1152/ajpgi.1999.276.5.G1279?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -