Show simple item record

dc.contributor.authorLie, Shervi
dc.contributor.authorRochet, Elise
dc.contributor.authorSegerdell, Erik
dc.contributor.authorMa, Yuefang
dc.contributor.authorAshander, Liam M
dc.contributor.authorShadforth, Audra MA
dc.contributor.authorBlenkinsop, Timothy A
dc.contributor.authorMichael, Michael Z
dc.contributor.authorAppukuttan, Binoy
dc.contributor.authorWilmot, Beth
dc.contributor.authorSmith, Justine R
dc.date.accessioned2019-09-08T23:46:31Z
dc.date.available2019-09-08T23:46:31Z
dc.date.issued2019
dc.identifier.issn1664-3224
dc.identifier.doi10.3389/fimmu.2019.00708
dc.identifier.urihttp://hdl.handle.net/10072/387023
dc.description.abstractOcular toxoplasmosis is the commonest clinical manifestation of infection with obligate intracellular parasite, Toxoplasma gondii. Active ocular toxoplasmosis is characterized by replication of T. gondii tachyzoites in the retina, with reactive inflammation. The multifunctional retinal pigment epithelium is a key target cell population for T. gondii. Since the global gene expression profile is germane to understanding molecular involvements of retinal pigment epithelial cells in ocular toxoplasmosis, we performed RNA-Sequencing (RNA-Seq) of human cells following infection with T. gondii tachyzoites. Primary cell isolates from eyes of cadaveric donors (n = 3), and the ARPE-19 human retinal pigment epithelial cell line, were infected for 24 h with GT-1 strain T. gondii tachyzoites (multiplicity of infection = 5) or incubated uninfected as control. Total and small RNA were extracted from cells and sequenced on the Illumina NextSeq 500 platform; results were aligned to the human hg19 reference sequence. Multidimensional scaling showed good separation between transcriptomes of infected and uninfected primary cell isolates, which were compared in edgeR software. This differential expression analysis revealed a sizeable response in the total RNA transcriptome—with significantly differentially expressed genes totaling 7,234 (28.9% of assigned transcripts)—but very limited changes in the small RNA transcriptome—totaling 30 (0.35% of assigned transcripts) and including 8 microRNA. Gene ontology and pathway enrichment analyses of differentially expressed total RNA in CAMERA software, identified a strong immunologic transcriptomic signature. We conducted RT-qPCR for 26 immune response-related protein-coding and long non-coding transcripts in epithelial cell isolates from different cadaveric donors (n = 3), extracted by a different isolation protocol but similarly infected with T. gondii, to confirm immunological activity of infected cells. For microRNA, increases in miR-146b and miR-212 were detected by RT-qPCR in 2 and 3 of these independent cell isolates. Biological network analysis in the InnateDB platform, including 735 annotated differentially expressed genes plus 2,046 first-order interactors, identified 10 contextural hubs and 5 subnetworks in the transcriptomic immune response of cells to T. gondii. Our observations provide a solid base for future studies of molecular and cellular interactions between T. gondii and the human retinal pigment epithelium to illuminate mechanisms of ocular toxoplasmosis.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherFrontiers Media
dc.relation.ispartofjournalFrontiers in immunology
dc.relation.ispartofvolume10
dc.subject.fieldofresearchImmunology
dc.subject.fieldofresearchMedical microbiology
dc.subject.fieldofresearchcode3204
dc.subject.fieldofresearchcode3207
dc.subject.keywordsScience & Technology
dc.subject.keywordsLife Sciences & Biomedicine
dc.subject.keywordshuman
dc.subject.keywordsretinal pigment epithelium
dc.titleImmunological Molecular Responses of Human Retinal Pigment Epithelial Cells to Infection With Toxoplasma gondii
dc.typeJournal article
dc.type.descriptionC1 - Articles
dcterms.bibliographicCitationLie, S; Rochet, E; Segerdell, E; Ma, Y; Ashander, LM; Shadforth, AMA; Blenkinsop, TA; Michael, MZ; Appukuttan, B; Wilmot, B; Smith, JR, Immunological Molecular Responses of Human Retinal Pigment Epithelial Cells to Infection With Toxoplasma gondii, Frontiers in immunology, 2019, 10
dcterms.dateAccepted2019-03-15
dcterms.licensehttp://creativecommons.org/licenses/by/4.0/
dc.date.updated2019-09-08T23:36:40Z
dc.description.versionVersion of Record (VoR)
gro.rights.copyright© 2019 Lie, Rochet, Segerdell, Ma, Ashander, Shadforth, Blenkinsop, Michael, Appukuttan, Wilmot and Smith. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
gro.hasfulltextFull Text
gro.griffith.authorShadforth, Audra


Files in this item

This item appears in the following Collection(s)

  • Journal articles
    Contains articles published by Griffith authors in scholarly journals.

Show simple item record