Varicella Zoster Virus Induces Nuclear Translocation of the Neurokinin-1 Receptor, Promoting Lamellipodia Formation and Viral Spread in Spinal Astrocytes

Bubak, Andrew N.; Como, Christina N.; Blackmon, Anna; Frietze, Seth; Mescher, Teresa; Jones, Dallas; Cohrs, Randall J.; Pauček, Petr; Baird, Nicholas L.; Nagel, Maria A.

doi:10.1093/infdis/jiy297

Cited by 12 publications

(11 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…HFLs were propagated as described above in an ibidi 24-well μ-Plate (ibidi) and analyzed for immunofluorescence as previously described [ 47 ]. Briefly, HFLs were fixed with 4% paraformaldehyde for 20 min at room temperature, followed by permeabilization with Triton-X (0.1%) for 10 min and blocked with normal donkey serum (10%) for 1 hour, then stained against ORF63 (1:1000), VZV gB (1:500) or respective isotype control for 16 h at 4°C, while secondary antibodies were incubated for 1 hour at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Varicella zoster virus productively infects human peripheral blood mononuclear cells to modulate expression of immunoinhibitory proteins and blocking PD-L1 enhances virus-specific CD8+ T cell effector function

et al. 2019

Self Cite

View full text Add to dashboard Cite

Varicella zoster virus (VZV) is a lymphotropic alpha-herpesvirinae subfamily member that produces varicella on primary infection and causes zoster, vascular disease and vision loss upon reactivation from latency. VZV-infected peripheral blood mononuclear cells (PBMCs) disseminate virus to distal organs to produce clinical disease. To assess immune evasion strategies elicited by VZV that may contribute to dissemination of infection, human PBMCs and VZV-specific CD8+ T cells (V-CD8+) were mock- or VZV-infected and analyzed for immunoinhibitory protein PD-1, PD-L1, PD-L2, CTLA-4, LAG-3 and TIM-3 expression using flow cytometry. All VZV-infected PBMCs (monocytes, NK, NKT, B cells, CD4+ and CD8+ T cells) and V-CD8+ showed significant elevations in PD-L1 expression compared to uninfected cells. VZV induced PD-L2 expression in B cells and V-CD8+. Only VZV-infected CD8+ T cells, NKT cells and V-CD8+ upregulated PD-1 expression, the immunoinhibitory receptor for PD-L1/PD-L2. VZV induced CTLA-4 expression only in V-CD8+ and no significant changes in LAG-3 or TIM-3 expression were observed in V-CD8+ or PBMC T cells. To test whether PD-L1, PD-L2 or CTLA-4 regulates V-CD8+ effector function, autologous PBMCs were VZV-infected and co-cultured with V-CD8+ cells in the presence of blocking antibodies against PD-L1, PD-L2 or CTLA-4; ELISAs revealed significant elevations in IFNγ only upon blocking of PD-L1. Together, these results identified additional immune cells that are permissive to VZV infection (monocytes, B cells and NKT cells); along with a novel mechanism for inhibiting CD8+ T cell effector function through induction of PD-L1 expression.

show abstract

Section: Methodsmentioning

confidence: 99%

Varicella zoster virus productively infects human peripheral blood mononuclear cells to modulate expression of immunoinhibitory proteins and blocking PD-L1 enhances virus-specific CD8+ T cell effector function

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…Cell-associated infections were used in all experiments to more accurately represent infection in vivo as described [ 15 ]. HA-sps and HA-hps were seeded at 5000 cells/cm 2 in basal astrocyte medium containing 2% fetal bovine serum (FBS), 1% astrocyte growth supplement, and 1% 100× penicillin-streptomycin (ScienCell).…”

Section: Methodsmentioning

confidence: 99%

“…After 24 h, medium was changed to basal astrocyte medium containing 0.1% FBS and 1% 100× penicillin-streptomycin and replenished every 72 h for 7 days, establishing quiescence. Quiescent HA-sps (qHA-sps) were co-cultivated with stocks of VZV-infected HA-sps (0.008 multiplicity of infection (MOI)) or mock-infected HA-sps and analyzed at 3 days post-infection (DPI) as described [ 15 ]; parallel experiments using quiescent HA-hps (qHA-hps) were also completed at the same MOI. This same protocol was followed for culturing and infecting primary human perineurial cells (HPNCs; ScienCell), which comprise the nerve-extrafascicular barrier.…”

Section: Methodsmentioning

confidence: 99%

“…RNA was extracted using the Direct-zol RNA MiniPrep kit (Zymo Research, Irvine) and reverse transcribed using the Transcriptor High Fidelity cDNA Synthesis Kit (Roche, Basel, Switzerland). Complementary DNA were analyzed with qPCR primers corresponding to each cytokine (IL-1β, Hs.PT.58.1518186; IL-2, Hs.PT.58.1142676; IL-4, Hs.PT.58.46539563g; IL-6, Hs.PT.58.40226675; IL-8, Hs.PT.58.39926886g; IL-10, Hs.PT.58.2807216; IL-12p70, Hs.PT.58.1687020; IL-13, Hs.PT.58.40619905.g; IFN-γ, Hs.PT.58.3781960; and TNF-α, Hs.PT.58.45380900; Integrated DNA Technologies, Coralville, IA) and glyceraldehyde-3-phosphate-dehydrogenase (GAPdH) as previously described [ 15 ]. Data were normalized to GAPdH and analyzed using the ∆∆ threshold cycle method.…”

Section: Methodsmentioning

confidence: 99%

“…In vitro studies have shown that astrocytes isolated from the human brain are permissive to VZV infection [ 13 ] and have decreased expression of glial fibrillary acidic protein (GFAP) [ 14 ]. Finally, a recent study of primary human spinal cord astrocytes revealed VZV induction of lamellipodia and filopodia that promotes viral spread in association with aberrant nuclear localization of the neurokinin-1 receptor [ 15 ]. In that study, a striking finding was the absence of morphological alterations indicative of astrocyte activation in the uninfected bystander astrocytes, suggesting the lack of “danger signals” associated with astrogliosis, cytokine production, and subsequent recruitment of inflammatory cells and microglia for viral clearance.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Varicella zoster virus differentially alters morphology and suppresses proinflammatory cytokines in primary human spinal cord and hippocampal astrocytes

Bubak

Como

Blackmon

et al. 2018

J Neuroinflammation

Self Cite

View full text Add to dashboard Cite

BackgroundVaricella zoster virus (VZV) is a ubiquitous alphaherpesvirus that produces varicella and zoster. VZV can infect multiple cell types in the spinal cord and brain, including astrocytes, producing myelopathy and encephalopathy. While studies of VZV-astrocyte interactions are sparse, a recent report showed that quiescent primary human spinal cord astrocytes (qHA-sps) did not appear activated morphologically during VZV infection. Since astrocytes play a critical role in host defenses during viral infections of the central nervous system, we examined the cytokine responses of qHA-sps and quiescent primary human hippocampal astrocytes (qHA-hps) to VZV infection in vitro, as well as the ability of conditioned supernatant to recruit immune cells.MethodsAt 3 days post-infection, mock- and VZV-infected qHA-sps and qHA-hps were examined for morphological changes by immunofluorescence antibody assay using antibodies directed against glial fibrillary acidic protein and VZV. Conditioned supernatants were analyzed for proinflammatory cytokines [interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, interferon-gamma, and tumor necrosis factor-α] using the Meso Scale Discovery multiplex ELISA platform. Finally, the ability of conditioned supernatants to attract peripheral blood mononuclear cells (PBMCs) was determined using a chemotaxis assay. Quiescent primary human perineurial cells (qHPNCs) served as a control for VZV-induced cytokine production and PBMC migration. To confirm that the astrocytes have the ability to increase cytokine secretion, qHA-sps and qHA-hps were treated with IL-1β and examined for morphological changes and IL-6 secretion.ResultsVZV-infected qHA-sps displayed extensive cellular processes, whereas VZV-infected qHA-hps became swollen and clustered together. Astrocytes had the capacity to secrete IL-6 in response to IL-1β. Compared to mock-infected cells, VZV-infected qHA-sps showed significantly reduced secretion of IL-2, IL-4, IL-6, IL-12p70, and IL-13, while VZV-infected qHA-hps showed significantly reduced IL-8 secretion. In contrast, levels of all 10 cytokines examined were significantly increased in VZV-infected qHPNCs. Consistent with these results, conditioned supernatant from VZV-infected qHPNCs, but not that from VZV-infected qHA-sps and qHA-hps, recruited PBMCs.ConclusionsVZV-infected qHA-sps and qHA-hps have distinct morphological alterations and patterns of proinflammatory cytokine suppression that could contribute to ineffective viral clearance in VZV myelopathy and encephalopathy, respectively.

show abstract

Trigeminal Postherpetic Neuralgia: From Pathophysiology to Treatment

Niemeyer,

Harlander-Locke,

Bubak

et al. 2024

Curr Pain Headache Rep

View full text Add to dashboard Cite

Purpose of Review Trigeminal postherpetic neuralgia (TG-PHN) is a neuropathic pain condition complicating herpes zoster (HZ) attributed to the trigeminal nerve. It poses significant challenges due to its persistent and debilitating nature. This review explores the clinical characteristics of TG-PHN, analyzes its pathophysiological underpinnings, and addresses existent and potential therapies. Recent Findings TG-PHN is one of the most common and complex PHN locations. It has distinguishing clinical and pathophysiological characteristics, starting with viral triggered injuries to the trigeminal ganglion (TG) and peripheral tissue and involving the ascending and descending brain modulation pathways. Current therapies include vaccines, oral and topical medications, and interventional approaches, like nerve blocks and neurostimulation. Summary This review covers TG-PHN’s clinical and physiological components, treatment options, and potential future targets for improved management. By exploring the complexities of this condition, we aim to contribute to developing more effective and targeted therapies for patients suffering from trigeminal PHN.

show abstract

Varicella Zoster Virus Induces Nuclear Translocation of the Neurokinin-1 Receptor, Promoting Lamellipodia Formation and Viral Spread in Spinal Astrocytes

Cited by 12 publications

References 43 publications

Varicella zoster virus productively infects human peripheral blood mononuclear cells to modulate expression of immunoinhibitory proteins and blocking PD-L1 enhances virus-specific CD8+ T cell effector function

Varicella zoster virus productively infects human peripheral blood mononuclear cells to modulate expression of immunoinhibitory proteins and blocking PD-L1 enhances virus-specific CD8+ T cell effector function

Varicella zoster virus differentially alters morphology and suppresses proinflammatory cytokines in primary human spinal cord and hippocampal astrocytes

Trigeminal Postherpetic Neuralgia: From Pathophysiology to Treatment

Contact Info

Product

Resources

About