Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells
Abstract Background Currently, no FDA-approved vaccines or treatments are available for Ebola virus disease (EVD), and therapy remains largely supportive. Ebola virus (EBOV) has broad tissue tropism and can infect a variety of cells including epithelial cells. Epithelial cells differ from most other...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-08-01
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| Series: | Virology Journal |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12985-018-1045-0 |
| _version_ | 1811219607506124800 |
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| author | Manasi Tamhankar Dawn M. Gerhardt Richard S. Bennett Nicole Murphy Peter B. Jahrling Jean L. Patterson |
| author_facet | Manasi Tamhankar Dawn M. Gerhardt Richard S. Bennett Nicole Murphy Peter B. Jahrling Jean L. Patterson |
| author_sort | Manasi Tamhankar |
| collection | DOAJ |
| description | Abstract Background Currently, no FDA-approved vaccines or treatments are available for Ebola virus disease (EVD), and therapy remains largely supportive. Ebola virus (EBOV) has broad tissue tropism and can infect a variety of cells including epithelial cells. Epithelial cells differ from most other cell types by their polarized phenotype and barrier function. In polarized cells, the apical and basolateral membrane domains are demarcated by tight junctions, and specialized sorting machinery, which results in a difference in composition between the two membrane domains. These specialized sorting functions can have important consequences for viral infections. Differential localization of a viral receptor can restrict virus entry to a particular membrane while polarized sorting can lead to a vectorial virus release. The present study investigated the impact of cell polarity on EBOV infection. Methods Characteristics of EBOV infection in polarized cells were evaluated in the polarized Caco-2 model grown on semipermeable transwells. Transepithelial resistance (TEER), which is a function of tight junctions, was used to assess epithelial cell polarization. EBOV infection was assessed with immunofluorescence microscopy and qPCR. Statistical significance was calculated using one-way ANOVA and significance was set at p < 0.05. Results Our data indicate that EBOV preferentially infects cells from the basolateral route, and this preference may be influenced by the resistance across the Caco-2 monolayer. Infection occurs without changes in cellular permeability. Further, our data show that basolateral infection bias may be dependent on polarized distribution of heparan sulfate, a known viral attachment factor. Treatment with iota-carrageenan, or heparin lyase, which interrupts viral interaction with cellular heparan sulfate, significantly reduced cell susceptibility to basolateral infection, likely by inhibiting virus attachment. Conclusions Our results show cell polarity has an impact on EBOV infection. EBOV preferentially infects polarized cells through the basolateral route. Access to heparan sulfate is an important factor during basolateral infection and blocking interaction of cellular heparan sulfate with virus leads to significant inhibition of basolateral infection in the polarized Caco-2 cell model. |
| first_indexed | 2024-04-12T07:29:37Z |
| format | Article |
| id | doaj.art-c18d663e89c4459eb2269f7b17fcd15e |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-12T07:29:37Z |
| publishDate | 2018-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-c18d663e89c4459eb2269f7b17fcd15e2022-12-22T03:42:07ZengBMCVirology Journal1743-422X2018-08-0115111210.1186/s12985-018-1045-0Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cellsManasi Tamhankar0Dawn M. Gerhardt1Richard S. Bennett2Nicole Murphy3Peter B. Jahrling4Jean L. Patterson5Department of Virology and Immunology, Texas Biomedical Research InstituteIntegrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthIntegrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthIntegrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthIntegrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of HealthDepartment of Virology and Immunology, Texas Biomedical Research InstituteAbstract Background Currently, no FDA-approved vaccines or treatments are available for Ebola virus disease (EVD), and therapy remains largely supportive. Ebola virus (EBOV) has broad tissue tropism and can infect a variety of cells including epithelial cells. Epithelial cells differ from most other cell types by their polarized phenotype and barrier function. In polarized cells, the apical and basolateral membrane domains are demarcated by tight junctions, and specialized sorting machinery, which results in a difference in composition between the two membrane domains. These specialized sorting functions can have important consequences for viral infections. Differential localization of a viral receptor can restrict virus entry to a particular membrane while polarized sorting can lead to a vectorial virus release. The present study investigated the impact of cell polarity on EBOV infection. Methods Characteristics of EBOV infection in polarized cells were evaluated in the polarized Caco-2 model grown on semipermeable transwells. Transepithelial resistance (TEER), which is a function of tight junctions, was used to assess epithelial cell polarization. EBOV infection was assessed with immunofluorescence microscopy and qPCR. Statistical significance was calculated using one-way ANOVA and significance was set at p < 0.05. Results Our data indicate that EBOV preferentially infects cells from the basolateral route, and this preference may be influenced by the resistance across the Caco-2 monolayer. Infection occurs without changes in cellular permeability. Further, our data show that basolateral infection bias may be dependent on polarized distribution of heparan sulfate, a known viral attachment factor. Treatment with iota-carrageenan, or heparin lyase, which interrupts viral interaction with cellular heparan sulfate, significantly reduced cell susceptibility to basolateral infection, likely by inhibiting virus attachment. Conclusions Our results show cell polarity has an impact on EBOV infection. EBOV preferentially infects polarized cells through the basolateral route. Access to heparan sulfate is an important factor during basolateral infection and blocking interaction of cellular heparan sulfate with virus leads to significant inhibition of basolateral infection in the polarized Caco-2 cell model.http://link.springer.com/article/10.1186/s12985-018-1045-0Ebola virusCaco-2Polarized cellsHeparan sulfate |
| spellingShingle | Manasi Tamhankar Dawn M. Gerhardt Richard S. Bennett Nicole Murphy Peter B. Jahrling Jean L. Patterson Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells Virology Journal Ebola virus Caco-2 Polarized cells Heparan sulfate |
| title | Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells |
| title_full | Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells |
| title_fullStr | Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells |
| title_full_unstemmed | Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells |
| title_short | Heparan sulfate is an important mediator of Ebola virus infection in polarized epithelial cells |
| title_sort | heparan sulfate is an important mediator of ebola virus infection in polarized epithelial cells |
| topic | Ebola virus Caco-2 Polarized cells Heparan sulfate |
| url | http://link.springer.com/article/10.1186/s12985-018-1045-0 |
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