scholarly journals Delayed induction of type I and III interferons mediates nasal epithelial cell permissiveness to SARS-CoV-2

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Catherine F. Hatton ◽  
Rachel A. Botting ◽  
Maria Emilia Dueñas ◽  
Iram J. Haq ◽  
Bernard Verdon ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Host Response ◽  
Cell Model ◽  
Viral Gene Expression ◽  
Cell Types ◽  
Nasal Delivery ◽  
Nasal Epithelium ◽  
Viral Gene ◽  
Type I ◽  
Nasal Epithelial Cell

AbstractThe nasal epithelium is a plausible entry point for SARS-CoV-2, a site of pathogenesis and transmission, and may initiate the host response to SARS-CoV-2. Antiviral interferon (IFN) responses are critical to outcome of SARS-CoV-2. Yet little is known about the interaction between SARS-CoV-2 and innate immunity in this tissue. Here we apply single-cell RNA sequencing and proteomics to a primary cell model of human nasal epithelium differentiated at air-liquid interface. SARS-CoV-2 demonstrates widespread tropism for nasal epithelial cell types. The host response is dominated by type I and III IFNs and interferon-stimulated gene products. This response is notably delayed in onset relative to viral gene expression and compared to other respiratory viruses. Nevertheless, once established, the paracrine IFN response begins to impact on SARS-CoV-2 replication. When provided prior to infection, recombinant IFNβ or IFNλ1 induces an efficient antiviral state that potently restricts SARS-CoV-2 viral replication, preserving epithelial barrier integrity. These data imply that the IFN-I/III response to SARS-CoV-2 initiates in the nasal airway and suggest nasal delivery of recombinant IFNs to be a potential chemoprophylactic strategy.

scholarly journals Delayed induction of type I and III interferons and nasal epithelial cell permissiveness to SARS-CoV-2

2021 ◽  
Author(s):  
Catherine F Hatton ◽  
Rachel A Botting ◽  
Maria Emilia Dueñas ◽  
Iram J Haq ◽  
Bernard Verdon ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Host Response ◽  
Cell Model ◽  
Viral Gene Expression ◽  
Cell Types ◽  
Nasal Delivery ◽  
Nasal Epithelium ◽  
Viral Gene ◽  
Type I ◽  
Nasal Epithelial Cell

AbstractThe nasal epithelium is a plausible entry point for SARS-CoV-2, a site of pathogenesis and transmission, and may initiate the host response to SARS-CoV-2. Antiviral interferon responses are critical to outcome of SARS-CoV-2. Yet little is known about the interaction between SARS-CoV-2 and innate immunity in this tissue. Here we applied single-cell RNA sequencing and proteomics to a primary cell model of human primary nasal epithelium differentiated at air-liquid interface. SARS-CoV-2 demonstrated widespread tropism for nasal epithelial cell types. The host response was dominated by type I and III IFNs and interferon-stimulated gene products. Nevertheless, this response was notably delayed in onset compared to viral gene expression, and thus failed to impact substantially on SARS-CoV-2 replication. However, when provided prior to infection, recombinant IFNβ or IFNλ1 induced an efficient antiviral state that potently restricted SARS-CoV-2 viral replication, preserving epithelial barrier integrity. These data suggest nasal delivery of recombinant IFNs to be a potential chemoprophylactic strategy.

scholarly journals Development of a Novel ex vivo Nasal Epithelial Cell Model Supporting Colonization With Human Nasal Microbiota

2019 ◽  
Vol 9 ◽  
Author(s):  
Derald D. Charles ◽  
James R. Fisher ◽  
Sarah M. Hoskinson ◽  
Audrie A. Medina-Colorado ◽  
Yi C. Shen ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Model ◽  
Ex Vivo ◽  
Nasal Epithelial Cell ◽  
Nasal Microbiota

scholarly journals Human cytomegalovirus transcriptome activity differs during replication in human fibroblast, epithelial and astrocyte cell lines

2012 ◽  
Vol 93 (5) ◽  
pp. 1046-1058 ◽  
Author(s):  
James C. Towler ◽  
Bahram Ebrahimi ◽  
Brian Lane ◽  
Andrew J. Davison ◽  
Derrick J. Dargan
Keyword(s):  
Gene Expression ◽  
Human Cytomegalovirus ◽  
Growth Kinetics ◽  
Viral Gene Expression ◽  
Cell Types ◽  
Viral Gene ◽  
Genome Replication ◽  
Cell Type ◽  
Low Passage ◽  
Temporal Expression Pattern

Broad cell tropism contributes to the pathogenesis of human cytomegalovirus (HCMV), but the extent to which cell type influences HCMV gene expression is unclear. A bespoke HCMV DNA microarray was used to monitor the transcriptome activity of the low passage Merlin strain of HCMV at 12, 24, 48 and 72 h post-infection, during a single round of replication in human fetal foreskin fibroblast cells (HFFF-2s), human retinal pigmented epithelial cells (RPE-1s) and human astrocytoma cells (U373MGs). In order to correlate transcriptome activity with concurrent biological responses, viral cytopathic effect, growth kinetics and genomic loads were examined in the three cell types. The temporal expression pattern of viral genes was broadly similar in HFFF-2s and RPE-1s, but dramatically different in U373MGs. Of the 165 known HCMV protein-coding genes, 41 and 48 were differentially regulated in RPE-1s and U373MGs, respectively, compared with HFFF-2s, and 22 of these were differentially regulated in both RPE-1s and U373MGs. In RPE-1s, all differentially regulated genes were downregulated, but, in U373MGs, some were down- and others upregulated. Differentially regulated genes were identified among the immediate-early, early, early late and true-late viral gene classes. Grouping of downregulated genes according to function at landmark stages of the replication cycle led to the identification of potential bottleneck stages (genome replication, virion assembly, and virion maturation and release) that may account for cell type-dependent viral growth kinetics. The possibility that cell type-specific differences in expressed cellular factors are responsible for modulation of viral gene expression is discussed.

scholarly journals Genome Size and Structure Determine Efficiency of Postinternalization Steps and Gene Transfer of Capsid-Modified Adenovirus Vectors in a Cell-Type-Specific Manner

2004 ◽  
Vol 78 (18) ◽  
pp. 10009-10022 ◽  
Author(s):  
Dmitry M. Shayakhmetov ◽  
Zong-Yi Li ◽  
Anuj Gaggar ◽  
Helen Gharwan ◽  
Vladimir Ternovoi ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Genome Size ◽  
First Generation ◽  
Viral Gene Expression ◽  
Protein Composition ◽  
Cell Types ◽  
Helper Virus ◽  
Viral Gene ◽  
Viral Genes ◽  
Size And Structure

ABSTRACT Adenovirus serotype 5 (Ad5) vectors containing Ad B-group fibers have become increasingly popular as gene transfer vectors because they efficiently transduce human cell types that are relatively refractory to Ad5 infection. So far, most B-group fiber-containing vectors have been first-generation vectors, deleted of E1 and/or E3 genes. Transduction with these vectors, however, results in viral gene expression and is associated with cytotoxicity and immune responses against transduced cells. To circumvent these problems, we developed fiber-chimeric Ad vectors devoid of all viral genes that were produced either by the homologous recombination of first-generation vectors or by using the Cre/lox-based helper virus system. In this study we compared early steps of infection between first-generation (35-kb genome) and Ad vectors devoid of all viral genes with genome sizes of 28 kb and 12.6 kb. All vectors possessed an Ad35-derived fiber knob domain, which uses CD46 as a primary attachment receptor. Using immortalized human hematopoietic cell lines and primary human CD34-positive hematopoietic cells, we found that the Ad genome size did not affect the efficiency of virus attachment to and internalization into cells. Furthermore, independently of the genome length and structure, all vectors migrated to the nucleus through late endosomal and lysosomal cellular compartments. However, the vector containing the short 12.6-kb genome was unable to efficiently escape from endosomes and deliver its DNA into the nucleus. Moreover, compared to other vectors, these Ad particles were less stable and had an abnormal capsid protein composition, including a lack of capsid-stabilizing protein IX. Our data indicate that the size and structure of the packaged viral genomes can affect the integrity of Ad particles, which in turn results in lower infectivity of Ad vectors.

scholarly journals Novel Nasal Epithelial Cell Markers of Parkinson’s Disease Identified Using Cells Treated with α-Synuclein Preformed Fibrils

2020 ◽  
Vol 9 (7) ◽  
pp. 2128 ◽  
Author(s):  
Hyojung Kim ◽  
Seok-Jae Kang ◽  
Young Mi Jo ◽  
Sanggyu Park ◽  
Seung Pil Yun ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Epithelial Cell ◽  
Cell Model ◽  
Sensory Perception ◽  
Olfactory Dysfunction ◽  
Clinical Samples ◽  
Chemical Stimulus ◽  
Nasal Epithelial Cell ◽  
Nasal Fluid

Parkinson’s disease (PD) is the most common neurodegenerative movement disorder, characterized by olfactory dysfunction in the early stages. α-Synuclein pathologies in the olfactory organs are shown to spread to the brain through the nose-brain axis. We first developed a nasal epithelial PD cellular model by treating RPMI-2650 cells with α-synuclein preformed fibrils (PFF). Upon uptake of PFF, RPMI-2650 cells showed mitochondrial proteome alteration and downregulation of parkin, which has previously been identified as a nasal biomarker of PD. Functional cluster analysis of differentially expressed genes in RPMI-2650 cells revealed various pathways affected by α-synuclein pathology, including the detection of chemical stimulus involved in sensory perception, olfactory receptor activity, and sensory perception of smell. Among genes that were most affected, we validated, by real-time quantitative PCR, the downregulation of MAP3K8, OR10A4, GRM2, OR51B6, and OR9A2, as well as upregulation of IFIT1B, EPN1, OR1D5, LCN, and OTOL1 in PFF-treated RPMI-2650 cells. Subsequent analyses of clinical samples showed a downregulation of OR10A4 and OR9A2 transcripts and an upregulation of IFIT1B in cells isolated from the nasal fluid of PD patients, as compared to those from the controls (cutoff value = 0.5689 for OR9A2, with 72.4% sensitivity and 75% specificity, and 1.4658 for IFIT1B, with 81.8% sensitivity and 77.8% specificity). Expression levels of these nasal PD markers were not altered in nasal fluid cells from SWEDD (scans without evidence of dopaminergic deficits) patients with PD-like motor symptoms. These nasal markers were significantly altered in patients of PD with hyposmia compared to the control hyposmic subjects. Our results validated the α-synuclein-treated nasal epithelial cell model to identify novel biomarkers for PD and suggest the utility of olfactory transcripts, along with olfactory dysfunction, in the diagnosis of PD.

Regulation of αIIbβ3-Signaling by Myomesin Family of Proteins.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3893-3893
Author(s):  
Kumar B. Reddy
Keyword(s):  
Cell Model ◽  
Cell Types ◽  
Positive Control ◽  
Variable Number ◽  
Model System ◽  
Type I ◽  
Integrin Receptors ◽  
Β3 Integrin ◽  
Cell Model System ◽  
Inside Out

Abstract Integrins are the type-I transmembrane proteins. They function as heterodimers of α and β subunits. The platelet integrin αIIbβ3 (GPIIb-IIIa) served as the model system in understanding the signaling across integrin receptors. αIIbβ3 exists in a resting state on circulating platelets and can be activated via several inside-out signaling pathways resulting in binding to its ligands, such as fibrinogen (Fg). The inside-out signaling appears to be regulated by several cytoskeletal and signaling proteins, which directly associate with the cytoplasmic tails of αIIbβ3. We have previously identified skelemin/myomesin-I as the cytoskeletal protein that interacted with the cytoplasmic tail of β3-integrins. Skelemin/myomesin-I is very closely related to two other members: myomesins-II and III. Myomesins I-III, along with other proteins such as titin and myosin light chain kinases, are members of a much larger superfamily of proteins. The hallmark of this family of proteins is the presence of a variable number of fibronectin (Fn)-like and immunoglobulin (Ig)-like motifs. Myomesin-I contains five Fn-motifs flanked by a total of seven Ig-like motifs. The interaction between β3-integrin and myomesin-I occurs via the C-terminal Ig motifs 3–7 of myomsin-I. In order to understand the functional consequence of this interaction between myomesin-I and β3-integrin, we measured the ligand binding properties of αIIbβ3 in a CHO cell model system. We found that expression of Ig motifs 3–7 increased Fg binding to the αIIbβ3, comparable to talin-head domain, used as the positive control. However, the expression of smaller and individual Ig motifs 4 and 5 resulted in still significantly higher Fg-binding to αIIbβ3. Ig motifs from myomesin-II also behaved in a similar fashion. We also found that myomesin-II is ubiquitously expressed in all the cell types tested, including platelets, suggesting myomesin-II is the likely candidate among various myomesin family members for the regulation of signaling across αIIbβ3. Our results suggest that the Ig motifs of the myomesin represent novel domains, which can regulate inside-out signaling across αIIbβ3.

scholarly journals Nuclear Localization of Tegument-Delivered pp71 in Human Cytomegalovirus-Infected Cells Is Facilitated by One or More Factors Present in Terminally Differentiated Fibroblasts

2010 ◽  
Vol 84 (19) ◽  
pp. 9853-9863 ◽  
Author(s):  
Rhiannon R. Penkert ◽  
Robert F. Kalejta
Keyword(s):  
Gene Expression ◽  
Human Cytomegalovirus ◽  
Viral Entry ◽  
Viral Gene Expression ◽  
Cell Types ◽  
Lytic Replication ◽  
Viral Gene ◽  
Differentiated Cells ◽  
Virion Envelope ◽  
Infected Cells

ABSTRACT Herpesviral virions contain a tegument layer that consists primarily of viral proteins. The delivery of fully functional proteins to infected cells upon virion envelope fusion to the plasma membrane allows herpesviruses to modulate cellular activities prior to viral gene expression. Certain tegument proteins can also regulate viral processes. For example, the pp71 tegument protein encoded by the UL82 gene of human cytomegalovirus (HCMV) stimulates viral immediate early (IE) gene expression and thus acts to initiate the productive lytic infectious cycle. In terminally differentiated fibroblasts infected with HCMV, tegument-delivered pp71 traffics to the nucleus and degrades the cellular transcriptional corepressor Daxx to initiate viral IE gene expression and lytic replication. However, when HCMV infects incompletely differentiated cells, tegument-delivered pp71 remains in the cytoplasm, allowing the nucleus-localized Daxx protein to silence viral IE gene expression and promote the establishment of a latent infection in certain cell types. We sought to determine whether undifferentiated cells block the trafficking of tegument-delivered pp71 to the nucleus or whether differentiated cells facilitate the nuclear transport of tegument-delivered pp71. Heterogenous cell fusion experiments demonstrated that tegument-delivered pp71 found in the cytoplasm of undifferentiated NT2 cells could be driven into the nucleus by one or more factors provided by fully differentiated fibroblasts. Our data raise the intriguing possibility that latency is the default program launched by HCMV upon viral entry into cells and that lytic infection is initiated only in certain (differentiated) cells that can facilitate the delivery of incoming pp71 to the nucleus.

SCD2-mediated monounsaturated fatty acid metabolism regulates cGAS-STING-dependent type I IFN in CD4+ T cells

2021 ◽  
Author(s):  
Toshio Kanno ◽  
Takahiro Nakajima ◽  
Satoru Yokoyama ◽  
Hikari Asou ◽  
Shigemi Sasamoto ◽  
...  
Keyword(s):  
T Cells ◽  
Fatty Acid ◽  
Lipid Metabolism ◽  
Fatty Acid Biosynthesis ◽  
Viral Gene Expression ◽  
Monounsaturated Fatty Acid ◽  
Viral Gene ◽  
Type I ◽  
Type I Ifn ◽  
Viral Response

Abstract Intimate interactions exist between the host lipid metabolism and viral responses. However, how acquired immune systems adapt lipid metabolism to meet demands and whether or not the metabolic rewiring confers a selective advantage in host immunity remains unclear. We found that viral infection attenuated the expression of genes related to lipid metabolism in CD4+ T cells, which in turn increased the anti-viral gene expression. The inhibition of the fatty acid synthesis pathway substantially increased the basal expression of anti-viral genes via the spontaneous production of type I interferon. Using a combination of CRISPR/Cas9-mediated genome editing technology and a global lipidomics analysis, we found that the decreased monounsaturated fatty acid by genetic deletion of Scd2 was crucial for the induction of an anti-viral response through the activation of cGAS-STING pathway. These findings demonstrate the novel relationship between fatty acid biosynthesis and type I IFN responses that enhances the anti-viral response.

scholarly journals Interleukin-36γ Is Elevated in Cervicovaginal Epithelial Cells in Women With Bacterial Vaginosis and In Vitro After Infection With Microbes Associated With Bacterial Vaginosis

2019 ◽  
Vol 221 (6) ◽  
pp. 983-988 ◽  
Author(s):  
Jameson K Gardner ◽  
Paweł Łaniewski ◽  
Anna Knight ◽  
Lisa B Haddad ◽  
Alison Swaims-Kohlmeier ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Bacterial Vaginosis ◽  
Host Response ◽  
Cell Model ◽  
Bacterial Species ◽  
Clinical Samples ◽  
Sexually Transmitted ◽  
3 Dimensional

Abstract In recent studies, the interleukin (IL)-36 cytokines were shown to be elevated in women with non-Lactobacillus-dominated vaginal microbiomes. In this study, we evaluated IL36G expression in clinical samples from women with and without bacterial vaginosis (BV) and a human 3-dimensional cervical epithelial cell model. IL36G expression was significantly elevated in cervicovaginal epithelial cells isolated from BV-positive women and corresponded with increased neutrophil counts relative to BV-negative women. In addition, specific BV-associated bacterial species as well as a polymicrobial cocktail significantly induced IL36G expression in vitro. These findings suggest that IL-36γ may exhibit an important function in the host response to BV and other sexually transmitted infections.

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