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HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study

BACKGROUND: How HIV-1 enter into the eyes remains obscure. We postulated that HIV-1 Tat protein can alter the expression of specific tight-junction proteins and disturb the blood retinal barrier, and contributes to HIV trafficking into the eyes. This study is to determine the effects of HIV-1 Tat pr...

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Autores principales: Bai, Ling, Zhang, Zhenping, Zhang, Hui, Li, Xiumei, Yu, Qiurong, Lin, Haotian, Yang, Wenhui
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2430207/
https://www.ncbi.nlm.nih.gov/pubmed/18538010
http://dx.doi.org/10.1186/1471-2334-8-77
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author Bai, Ling
Zhang, Zhenping
Zhang, Hui
Li, Xiumei
Yu, Qiurong
Lin, Haotian
Yang, Wenhui
author_facet Bai, Ling
Zhang, Zhenping
Zhang, Hui
Li, Xiumei
Yu, Qiurong
Lin, Haotian
Yang, Wenhui
author_sort Bai, Ling
collection PubMed
description BACKGROUND: How HIV-1 enter into the eyes remains obscure. We postulated that HIV-1 Tat protein can alter the expression of specific tight-junction proteins and disturb the blood retinal barrier, and contributes to HIV trafficking into the eyes. This study is to determine the effects of HIV-1 Tat proteins on the barrier function and tight-junction protein expression of retinal pigment epithelial cell (RPE). METHODS: A human RPE cell line (D407) cultured on microporous filter-supports was used. After treating with HIV-1 Tat protein, transepithelial electrical resistance (TER) of confluent RPE cells was measured by epithelial voltmeter. The permeability of the RPE cells to sodium fluorescein was measured. The expressions of the occludin and claudins were determined by real-time polymerase chain reaction, immunofluorescence, and Western blot analysis. Activation of ERK1/2 was detected by Western blot analysis with specific antiphospho protein antibodies. NF-κB DNA binding activity was determined by transcription factor assay. Specific pharmacologic inhibitors directed against the MAPKs were used to analyze the signaling involved in barrier destruction of RPE cells exposed to HIV-1 Tat. RESULTS: Treating cultured human retinal pigment epithelial cells with 100 nM Tat for 24 hours increased the permeability and decreased the TER of the epithelial monolayer. HIV-1 Tat also disrupted and downregulated the tight-junction proteins claudin-1, claudin-3, and claudin-4 in these cells, whereas claudin-2 was upregulated, and the expression of occludin was unaffected. HIV-1 Tat protein also induced activation of ERK1/2 and NF-κB. HIV-1 Tat protein induced barrier destruction, changes in expression of TJs, and activation of ERK1/2 and NF-κB were abrogated by inhibitor of ERK1/2 and NF-κB. CONCLUSION: HIV-1 Tat protein causes increases in the paracellular permeability of RPE cells in vitro concomitant with changes in expression of certain transmembrane proteins associated with the tight junction. The effects of HIV-1 Tat on barrier function of the RPE may be mediated by ERK MAPK and NF-κB activation, which may represent potential targets for novel therapeutic approaches for the retinopathy induced by HIV infection.
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spelling pubmed-24302072008-06-17 HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study Bai, Ling Zhang, Zhenping Zhang, Hui Li, Xiumei Yu, Qiurong Lin, Haotian Yang, Wenhui BMC Infect Dis Research Article BACKGROUND: How HIV-1 enter into the eyes remains obscure. We postulated that HIV-1 Tat protein can alter the expression of specific tight-junction proteins and disturb the blood retinal barrier, and contributes to HIV trafficking into the eyes. This study is to determine the effects of HIV-1 Tat proteins on the barrier function and tight-junction protein expression of retinal pigment epithelial cell (RPE). METHODS: A human RPE cell line (D407) cultured on microporous filter-supports was used. After treating with HIV-1 Tat protein, transepithelial electrical resistance (TER) of confluent RPE cells was measured by epithelial voltmeter. The permeability of the RPE cells to sodium fluorescein was measured. The expressions of the occludin and claudins were determined by real-time polymerase chain reaction, immunofluorescence, and Western blot analysis. Activation of ERK1/2 was detected by Western blot analysis with specific antiphospho protein antibodies. NF-κB DNA binding activity was determined by transcription factor assay. Specific pharmacologic inhibitors directed against the MAPKs were used to analyze the signaling involved in barrier destruction of RPE cells exposed to HIV-1 Tat. RESULTS: Treating cultured human retinal pigment epithelial cells with 100 nM Tat for 24 hours increased the permeability and decreased the TER of the epithelial monolayer. HIV-1 Tat also disrupted and downregulated the tight-junction proteins claudin-1, claudin-3, and claudin-4 in these cells, whereas claudin-2 was upregulated, and the expression of occludin was unaffected. HIV-1 Tat protein also induced activation of ERK1/2 and NF-κB. HIV-1 Tat protein induced barrier destruction, changes in expression of TJs, and activation of ERK1/2 and NF-κB were abrogated by inhibitor of ERK1/2 and NF-κB. CONCLUSION: HIV-1 Tat protein causes increases in the paracellular permeability of RPE cells in vitro concomitant with changes in expression of certain transmembrane proteins associated with the tight junction. The effects of HIV-1 Tat on barrier function of the RPE may be mediated by ERK MAPK and NF-κB activation, which may represent potential targets for novel therapeutic approaches for the retinopathy induced by HIV infection. BioMed Central 2008-06-06 /pmc/articles/PMC2430207/ /pubmed/18538010 http://dx.doi.org/10.1186/1471-2334-8-77 Text en Copyright © 2008 Bai et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Bai, Ling
Zhang, Zhenping
Zhang, Hui
Li, Xiumei
Yu, Qiurong
Lin, Haotian
Yang, Wenhui
HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title_full HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title_fullStr HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title_full_unstemmed HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title_short HIV-1 Tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
title_sort hiv-1 tat protein alter the tight junction integrity and function of retinal pigment epithelium: an in vitro study
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2430207/
https://www.ncbi.nlm.nih.gov/pubmed/18538010
http://dx.doi.org/10.1186/1471-2334-8-77
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