Release of Intracellular Calcium Stores Facilitates Coxsackievirus Entry into Polarized Endothelial Cells

Bozym, Rebecca A.; Morosky, Stefanie; Kim, Kwang S.; Cherry, Sara; Coyne, Carolyn B.

doi:10.1371/journal.ppat.1001135

Cited by 54 publications

(67 citation statements)

References 55 publications

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“…A wide range of cellular receptors has been observed in human EVs (reviewed by Merilahti et al, 2012). Although we did not examine the binding processes of EVs to the hCMEC/D3 cell surface, there is a large body of earlier experimental evidence to suggest that the intertypic variations in hCMEC/D3 susceptibility to EVs can be attributed to their propensity for using a wide range of receptors and internalization processes (Coyne et al, 2007;Bozym et al, 2010;Ylipaasto et al, 2010). For example, E-1 stands apart within the susceptibility spectrum of hCMEC/D3 cells to EV infection -a pattern that may be related to the fact that it is the only type known to bind integrin a 2 b 1 (Bergelson et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

“…These data suggest a role of infected leukocytes in EV dissemination to the CNS through a 'Trojan horse' process. In vitro studies showed the susceptibility to different EVs of human vascular endothelial cells of different tissue origins (Conaldi et al, 1997;Saijets et al, 2003;Zanone et al, 2003;Liang et al, 2004;Bozym et al, 2010;Ylipaasto et al, 2010). In addition, PV-1 and CV-B3 induced different cell signalling and endocytosis pathways in human brain microvascular endothelial cells, which was suggestive of possible variations in BBB crossing between EV types (Coyne et al, 2007;Bozym et al, 2010).…”

Section: Introductionmentioning

confidence: 97%

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Differential permissivity of human cerebrovascular endothelial cells to enterovirus infection and specificities of serotype EV-A71 in crossing an in vitro model of the human blood–brain barrier

Volle

Archimbaud

Couraud

et al. 2015

Journal of General Virology

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Human cerebral microvascular endothelial cells (hCMEC/D3 cell line) form a steady polarized barrier when cultured in vitro on a permeable membrane. Their susceptibility to enterovirus (EV) strains was analysed to investigate how these viruses may cross the blood-brain barrier. A sample of 88 virus strains was selected on phylogenetic features amongst 43 epidemiologically relevant types of the four EV species A-D. The EV-A71 genome was replicated at substantial rates, whilst the infectious virus was released at extremely low but sustained rates at both barrier sides for at least 4 days. EV-A71 antigens were detected in a limited number of cells. The properties of the endothelial barrier (structure and permeability) remained intact throughout infection. The chronic EV-A71 infection was in sharp contrast to the productive infection of cytolytic EVs (e.g. echoviruses E-6 and E-30). The hCMEC/D3 barriers infected with the latter EVs exhibited elevated proportions of apoptotic and necrotic cells, which resulted in major injuries to the endothelial barriers with a dramatic increase of paracellular permeability and virus crossing to the abluminal side. The following intracellular rearrangements were also seen: early destruction of the actin cytoskeleton, remodelling of intracellular membranes and reorganization of the mitochondrion network in a small cluster near the perinuclear space.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Differential permissivity of human cerebrovascular endothelial cells to enterovirus infection and specificities of serotype EV-A71 in crossing an in vitro model of the human blood–brain barrier

Volle

Archimbaud

Couraud

et al. 2015

Journal of General Virology

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“…In these cells, CVB enters by a lipid raft-dependent, dynamin-independent mechanism that combines features of caveolar endocytosis and macropinocytosis (6,11). In contrast, CVB entry into polarized endothelial cells requires caveolar endocytosis, dynamin II, and unidentified Src family tyrosine kinases (SFKs) (13). In nonpolarized HeLa cells, dynamin II and lipid rafts are necessary for CVB entry, while clathrin and caveolin-1 are not (14).…”

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confidence: 99%

Lipid Raft- and Src Family Kinase-Dependent Entry of Coxsackievirus B into Human Placental Trophoblasts

Delorme-Axford

Sadovsky

Coyne

2013

J Virol

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bMaternal-fetal transmission of group B coxsackieviruses (CVB) during pregnancy has been associated with a number of diverse pathological outcomes, including hydrops fetalis, fetal myocarditis, meningoencephalitis, neurodevelopmental delays, congenital skin lesions, miscarriage, and/or stillbirth. Throughout pregnancy, the placenta forms a critical antimicrobial protective barrier at the maternal-fetal interface. Despite the severity of diseases accompanying fetal CVB infections, little is known regarding the strategies used by CVB to gain entry into placental trophoblasts. Here we used both a trophoblast cell line and primary human trophoblasts to demonstrate the mechanism by which CVB gains entry into polarized placental trophoblasts. Our studies revealed that the kinetics of CVB entry into placental trophoblasts are similar to those previously described for polarized intestinal epithelial cells. Likewise, CVB entry into placental trophoblasts requires decay-accelerating factor (DAF) binding and involves relocalization of the virus from the apical surface to intercellular tight junctions. In contrast, we have identified a divergent mechanism for CVB entry into polarized trophoblasts that is clathrin, caveolin-1, and dynamin II independent but requires intact lipid rafts. In addition, we found that members of the Src family of tyrosine kinases were required for CVB entry. Our studies highlight the complexity of viral entry into human placental trophoblasts and may serve as a model for mechanisms used by diverse pathogens to penetrate the placental barrier.

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“…These nonreceptor tyrosine kinases readily autophosphorylate in bacteria and in vitro (131,132), suggesting that such substrates could be prepared in high yield for Abl-, Fyn-, and Lck-focused phosphatase assays. CVB3 infection also mobilizes Ca 2ϩ stores (133), and the Ca 2ϩ -activated phosphatase calcineurin disinhibits NFAT, a transcription factor whose activation in T cells promotes myocarditis (134). A calcineurinfocused NFAT phosphatase assay would enable a further elaboration of the host-cell signaling networks perturbed by CVB3.…”

Section: Discussionmentioning

confidence: 99%

“…The activity of subcellular phosphatases is then quantified by phospho-ELISA using a panel of recognized full-length phosphoproteins. Building upon our past success with phosphorylated MAPKs (29), we add three new phosphosubstrates-phospho-MK2 (Thr 334 ), phospho-CREB (Ser 133 ), and phospho-STAT1 (Tyr 701 )-each with distinct patterns of localization and targeting by protein phosphatase enzymes (Fig. 1).…”

mentioning

confidence: 99%

Profiling Subcellular Protein Phosphatase Responses to Coxsackievirus B3 Infection of Cardiomyocytes

Shah¹,

Smolko²,

Kinicki³

et al. 2017

Molecular & Cellular Proteomics

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, and the nucleus. The extracts contain all major classes of protein phosphatases and catalyze dephosphorylation of plate-bound phosphosubstrates in a microtiter format, with cellular activity quantified at the end point by phosphospecific ELISA. The platform is optimized for six phosphosubstrates (ERK2, JNK1, p38␣, MK2, CREB, and STAT1) and measures specific activities from extracts of fewer than 50,000 cells. The assay was exploited to examine viral and antiviral signaling in AC16 cardiomyocytes, which we show can be engineered to serve as susceptible and permissive hosts for CVB3. Phosphatase responses were profiled in these cells by completing a full-factorial experiment for CVB3 infection and type I/II interferon signaling. Over 850 functional measurements revealed several independent, subcellular changes in specific phosphatase activities. During CVB3 infection, we found that type I interferon signaling increases subcellular JNK1 phosphatase activity, inhibiting nuclear JNK1 activity that otherwise promotes viral protein synthesis in the infected host cell. Our assay provides a high-throughput way to capture perturbations in important negative regulators of intracellular signaltransduction networks. Molecular & Cellular Proteomics 16:

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Release of Intracellular Calcium Stores Facilitates Coxsackievirus Entry into Polarized Endothelial Cells

Cited by 54 publications

References 55 publications

Differential permissivity of human cerebrovascular endothelial cells to enterovirus infection and specificities of serotype EV-A71 in crossing an in vitro model of the human blood–brain barrier

Differential permissivity of human cerebrovascular endothelial cells to enterovirus infection and specificities of serotype EV-A71 in crossing an in vitro model of the human blood–brain barrier

Lipid Raft- and Src Family Kinase-Dependent Entry of Coxsackievirus B into Human Placental Trophoblasts

Profiling Subcellular Protein Phosphatase Responses to Coxsackievirus B3 Infection of Cardiomyocytes

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