title: Suppressor of cytokine signalling protein SOCS1 and UBP43 regulate the expression of type I interferon-stimulated genes in human microvascular endothelial cells infected with Rickettsia conorii.
abstract: Rickettsia conorii, the causative agent of Mediterranean spotted fever, preferentially infects human microvascular endothelium and activates pro-inflammatory innate immune responses as evidenced by enhanced expression and secretion of cytokines and chemokines. Our recent studies reveal that human microvascular endothelial cells (HMECs) infected with R. conorii also launch 'antiviral' host defence mechanisms typically governed by type I interferons. To summarize, infected HMECs secrete IFN-β to activate STAT1 in an autocrine/paracrine manner and display increased expression of IFN-stimulated genes, for example ISG15, which in turn activate innate responses to interfere with intracellular replication of rickettsiae. We now present evidence that UBP43 and SOCS1, known negative regulators of JAK/STAT signalling, are also induced in R. conorii-infected HMECs, of which UBP43 but not SOCS1 functions to negatively regulate STAT1 activation. Interestingly, UBP43 induction is almost completely abolished in the presence of IFN-β-neutralizing antibody, implicating an important role for UBP43 as a feedback inhibitor for IFN-β-mediated STAT1 activation. In contrast, SOCS1 expression is only partially affected by IFN-β neutralization, implicating potential involvement of as-yet-unidentified IFN-independent mechanism(s) in SOCS1 induction during R. conorii infection. A number of IFN-stimulated genes, including ISG15, OAS1, MX1, IRF1, IRF9 and TAP1 are also induced in an IFN-β-dependent manner, whereas GBP1 remains unaffected by IFN-β neutralization. Increased STAT1 phosphorylation in HMECs subjected to UBP43 knockdown led to transcriptional activation of OAS1, MX1 and GBP1, confirming the negative regulatory role of UBP43. Although IRF1, IRF9 and TAP1 were induced by IFN-β, siRNA-mediated silencing of UBP43 or SOCS1 did not significantly affect their transcriptional activation. Expression of ISG15 was, however, increased in HMECs transfected with siRNA for UBP43 and SOCS1. Thus, unique regulatory patterns of induced expression of UBP43, SOCS1 and IFN-stimulated genes represent pathogen-specific responses underlying IFN-β-mediated host endothelial signalling during the pathogenesis of spotted fever group rickettsiosis.
authors.affiliation: Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.|Department of Pathology and Laboratory Medicine, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.
authors.forename: Punsiri M
authors.initials: PM
authors.lastname: Colonne
authors.affiliation: Department of Pathology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA.
authors.forename: Abha
authors.initials: A
authors.lastname: Sahni
authors.affiliation: Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, TX 77555, USA.|Department of Pathology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA.
authors.forename: Sanjeev K
authors.initials: SK
authors.lastname: Sahni
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D010450:Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000382:microbiology*; D005786:Gene Expression Regulation / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D055785:Gene Knockdown Techniques; D006801:Humans; D007370:Interferon Type I / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020114:Rickettsia conorii / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D050826:Suppressor of Cytokine Signaling Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase; D014709:Vero Cells
chemical_list: D007370:Interferon Type I; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D050826:Suppressor of Cytokine Signaling Proteins; D010450:Endopeptidases; C485560:USP18 protein, human; D043222:Ubiquitin Thiolesterase
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