scholarly journals Conjunctival epithelial cells resist productive SARS-CoV-2 infection

2021 ◽  
Author(s):  
Majlinda Lako ◽  
Robert M Jackson ◽  
Catherine F Hatton ◽  
Jarmila S Spegarova ◽  
Maria Georgiou ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Viral Entry ◽  
Ocular Surface ◽  
Cell Types ◽  
Productive Infection ◽  
Conjunctival Epithelium ◽  
Genome Expression ◽  
Open Question ◽  
Air Liquid Interface ◽  
Conjunctival Epithelial Cells

Although tropism of SARS-CoV-2 for respiratory tract epithelial cells is well established, an open question is whether the conjunctival epithelium is also a target for SARS-CoV-2. Conjunctival epithelial cells, which express viral entry receptors ACE2 and TMPRSS2, constitute the largest exposed epithelium of the ocular surface tissue, and may represent a relevant viral entry route. To address this question, we generated an organotypic air-liquid-interface model of conjunctival epithelium, composed of progenitor, basal and superficial epithelial cells and fibroblasts, which could be maintained successfully up to day 75 of differentiation. Using single-cell RNA Seq, with complementary imaging and virological assays, we observed that while all conjunctival cell types were permissive to SARS-CoV-2 genome expression, a productive infection did not ensue. The early innate immune response to SARS-CoV-2 infection in conjunctival cells was characterised by a robust autocrine and paracrine NF-Kβ activity, without activation of antiviral interferon signalling. Collectively, these data enrich our understanding of SARS-CoV-2 infection at the human ocular surface, with potential implications for the design of preventive strategies and conjunctival transplants.

scholarly journals Coculture of autologous limbal and conjunctival epithelial cells to treat severe ocular surface disorders: Long-term survival analysis

2013 ◽  
Vol 61 (5) ◽  
pp. 202 ◽  
Author(s):  
VirenderS Sangwan ◽  
Kunjal Sejpal ◽  
SandhyaV Subramaniam ◽  
GeetaK Vemuganti ◽  
Anees Fatima ◽  
...  
Keyword(s):  
Survival Analysis ◽  
Epithelial Cells ◽  
Ocular Surface ◽  
Term Survival ◽  
Long Term Survival ◽  
Conjunctival Epithelial Cells

Roles of human β-defensins in innate immune defense at the ocular surface: arming and alarming corneal and conjunctival epithelial cells

2010 ◽  
Vol 134 (1) ◽  
pp. 59-73 ◽  
Author(s):  
Fabian Garreis ◽  
Thomas Schlorf ◽  
Dieter Worlitzsch ◽  
Philipp Steven ◽  
Lars Bräuer ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Ocular Surface ◽  
Immune Defense ◽  
Innate Immune ◽  
Innate Immune Defense ◽  
Conjunctival Epithelial Cells

scholarly journals Material Characterisation and Stratification of Conjunctival Epithelial Cells on Electrospun Poly(ε-Caprolactone) Fibres Loaded with Decellularised Tissue Matrices

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 318
Author(s):  
Lucy A. Bosworth ◽  
Kyle G. Doherty ◽  
James D. Hsuan ◽  
Samuel P. Cray ◽  
Raechelle A. D’Sa ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Fibre Diameter ◽  
Advanced Therapy Medicinal Product ◽  
Urinary Bladder Matrix ◽  
Advanced Therapy ◽  
Cell Layers ◽  
Intestinal Submucosa ◽  
Tissue Grafts ◽  
Air Liquid Interface ◽  
Conjunctival Epithelial Cells

The conjunctiva, an under-researched yet incredibly important tissue, plays key roles in providing protection to the eye and maintaining homeostasis of its ocular surface. Multiple diseases can impair conjunctival function leading to severe consequences that require surgical intervention. Small conjunctival defects can be repaired relatively easily, but larger defects rely on tissue grafts which generally do not provide adequate healing. A tissue engineering approach involving a biomaterial substrate capable of supporting a stratified epithelium with embedded, mucin-secreting goblet cells offers a potential solution. As a first step, this study aimed to induce stratification of human conjunctival epithelial cells cultured on electrospun scaffolds composed from poly(ε-caprolactone) (PCL) and decellularised tissue matrix (small intestinal submucosa (SIS) or urinary bladder matrix (UBM)) and held at the air/liquid interface. Stratification, up to 5 cell layers, occurred more frequently on scaffolds containing PCL + UBM. Incorporation of these decellularised tissue matrices also impacted material properties, with significant changes occurring to their fibre diameter, tensile properties, and chemical composition throughout the scaffold structure compared to PCL alone. These matrix containing scaffolds warrant further long-term investigation as a potential advanced therapy medicinal product for conjunctiva repair and regeneration.

scholarly journals Cells of the human respiratory tract support the replication of pathogenic Old World orthohantavirus Puumala

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Stefan Hägele ◽  
Christian Nusshag ◽  
Alexander Müller ◽  
Alexandra Baumann ◽  
Martin Zeier ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cells ◽  
Respiratory Tract ◽  
Cell Types ◽  
Airway Epithelial Cells ◽  
Productive Infection ◽  
Human Respiratory Tract ◽  
Old World ◽  
Pulmonary Manifestations ◽  
Kidney Involvement

Abstract Background Transmission of all known pathogenic orthohantaviruses (family Hantaviridae) usually occurs via inhalation of aerosols contaminated with viral particles derived from infected rodents and organ manifestation of infections is characterized by lung and kidney involvement. Orthohantaviruses found in Eurasia cause hemorrhagic fever with renal syndrome (HFRS) and New World orthohantaviruses cause hantavirus cardiopulmonary syndrome (HCPS). However, cases of infection with Old World orthohantaviruses with severe pulmonary manifestations have also been observed. Therefore, human airway cells may represent initial targets for orthohantavirus infection and may also play a role in the pathogenesis of infections with Eurasian orthohantaviruses. Methods We analyzed the permissiveness of primary endothelial cells of the human pulmonary microvasculature and of primary human epithelial cells derived from bronchi, bronchioles and alveoli for Old World orthohantavirus Puumala virus (PUUV) in vitro. In addition, we examined the expression of orthohantaviral receptors in these cell types. To minimize donor-specific effects, cells from two different donors were tested for each cell type. Results Productive infection with PUUV was observed for endothelial cells of the microvasculature and for the three tested epithelial cell types derived from different sites of the respiratory tract. Interestingly, infection and particle release were also detected in bronchial and bronchiolar epithelial cells although expression of the orthohantaviral receptor integrin β3 was not detectable in these cell types. In addition, replication kinetics and viral release demonstrate enormous donor-specific variations. Conclusions The human respiratory epithelium is among the first targets of orthohantaviral infection and may contribute to virus replication, dissemination and pathogenesis of HFRS-causing orthohantaviruses. Differences in initial pulmonary infection due to donor-specific factors may play a role in the observed broad variance of severity and symptoms of orthohantavirus disease in patients. The absence of detectable levels of integrin αVβ3 surface expression on bronchial and small airway epithelial cells indicates an alternate mode of orthohantaviral entry in these cells that is independent from integrin β3.

Differential regulation of centrin genes during ciliogenesis in human tracheal epithelial cells

1998 ◽  
Vol 275 (6) ◽  
pp. L1145-L1156 ◽  
Author(s):  
Michel LeDizet ◽  
James C. Beck ◽  
Walter E. Finkbeiner
Keyword(s):  
Epithelial Cells ◽  
Calcium Binding ◽  
Cell Types ◽  
Differential Regulation ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Tracheal Epithelial Cells ◽  
Tracheal Epithelial ◽  
Air Liquid Interface

Centrins are small calcium-binding proteins found in a variety of cell types, often in association with microtubule-organizing centers. Here we present results regarding the expression of centrins during the in vitro differentiation of human tracheal epithelial cells. When grown at an air-liquid interface, these cells differentiate into mucus-secreting cells or undergo ciliogenesis. In immunofluorescence and immunoelectron microscopy experiments, an anti-centrin antibody stained exclusively the basal bodies of the ciliated cells. There was no staining over the axonemes or the striated rootlets. Northern blots and RT-PCR analysis of the three known human centrin genes showed that these genes have distinct patterns of expression during the growth and differentiation of human tracheal epithelial cells. Centrin-1 is never transcribed. Centrin-2 mRNA is present at all times, and its concentration increases when ciliogenesis occurs. Centrin-3 mRNA is found at a constant level throughout the entire process. This differential regulation suggests that centrins are not interchangeable but instead have unique functions.

scholarly journals In Vitro Studies of Mechanisms of Lung Injury in the Rodent

1991 ◽  
Vol 19 (4_part_1) ◽  
pp. 419-427 ◽  
Author(s):  
Leah A. Cohn ◽  
Kenneth B. Adler
Keyword(s):  
Epithelial Cells ◽  
Cell Types ◽  
Lung Cells ◽  
Serum Free Medium ◽  
Collagen Gels ◽  
Respiratory Epithelial Cells ◽  
Respiratory Epithelial ◽  
Air Liquid Interface

In order to better define the responses of lung cells to potentially pathogenic insults, primary cell cultures of dissociated respiratory epithelial cells have been established. These epithelial cells have been obtained from various areas of the respiratory tract ranging from the trachea to the alveolus and the cultures have been demonstrated to mimic the differentiated state of these cell types as observed in situ. Several procedures which enhance the differentiated state have been evaluated, which include maintenance on more physiologically-relevant substrata, such as collagen gels, use of defined serum-free medium and use of air/liquid interface systems. These approaches have allowed intracellular responses of respiratory epithelium to toxic insult to be better defined.

scholarly journals Comparison of cytotoxicity effects induced by four different types of nanoparticles in human corneal and conjunctival epithelial cells

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Xiangzhe Li ◽  
Boram Kang ◽  
Youngsub Eom ◽  
Jingxiang Zhong ◽  
Hyung Keun Lee ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Ocular Surface ◽  
Protective Role ◽  
Toxic Effects ◽  
Chemical Components ◽  
Different Types ◽  
Cytotoxicity Effects ◽  
The Impact ◽  
Conjunctival Epithelial Cells

AbstractThe impact of particulate matter (PM) on ocular surface health has attracted increased attention in recent years. Previous studies have reported that differences in the chemical composition of PM can affect the toxicological response. However, available information on the toxic effects of chemical components of PM on the ocular surface is insufficient. In this paper, we aimed to investigate the toxicity effects of chemical components of PM on the ocular surface, focusing on the effects of four different types of nanoparticles (NPs) in human corneal epithelial cells (HCECs) and human conjunctival epithelial cells (HCjECs), which include titanium dioxide (TiO2), carbon black (CB), zinc dioxide (ZnO), and silicon dioxide (SiO2). We found that the in vitro cytotoxic effects of CB, ZnO, and SiO2 NPs are dependent on particle properties and cell type as well as the exposure concentration and time. Here, the order of increasing toxicity was SiO2 → CB → ZnO, while TiO2 demonstrated no toxicity. Moreover, toxic effects appearing more severe in HCECs than HCjECs. Reactive oxygen species (ROS)-mediated oxidative stress plays a key role in the toxicity of these three NPs in HCECs and HCjECs, leading to apoptosis and mitochondrial damage, which are also important contributors to aging. Sirtuin1 (SIRT1) as an NAD+-dependent protein deacetylase that seems to play a potential protective role in this process. These findings implied that ROS and/or SIRT1 may become a potential target of clinical treatment of PM- or NP-related ocular surface diseases.

scholarly journals Calcium signals and calpain-dependent necrosis are essential for release of coxsackievirus B from polarized intestinal epithelial cells

2011 ◽  
Vol 22 (17) ◽  
pp. 3010-3021 ◽  
Author(s):  
Rebecca A. Bozym ◽  
Kunal Patel ◽  
Carl White ◽  
King-Ho Cheung ◽  
Jeffrey M. Bergelson ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Viral Entry ◽  
Calcium Release ◽  
Cell Types ◽  
Junctional Complex ◽  
Intestinal Epithelial ◽  
Viral Pathogens ◽  
Coxsackievirus B ◽  
Calpain 2 ◽  
Trisphosphate Receptor

Coxsackievirus B (CVB), a member of the enterovirus family, targets the polarized epithelial cells lining the intestinal tract early in infection. Although the polarized epithelium functions as a protective barrier, this barrier is likely exploited by CVB to promote viral entry and subsequent egress. Here we show that, in contrast to nonpolarized cells, CVB-infected polarized intestinal Caco-2 cells undergo nonapoptotic necrotic cell death triggered by inositol 1,4,5-trisphosphate receptor–dependent calcium release. We further show that CVB-induced cellular necrosis depends on the Ca2+-activated protease calpain-2 and that this protease is involved in CVB-induced disruption of the junctional complex and rearrangements of the actin cytoskeleton. Our study illustrates the cell signaling pathways hijacked by CVB, and perhaps other viral pathogens, to promote their replication and spread in polarized cell types.

scholarly journals Apricot Kernel Extract and Amygdalin Inhibit Urban Particulate Matter-Induced Keratoconjunctivitis Sicca

Molecules ◽  
2019 ◽  
Vol 24 (3) ◽  
pp. 650 ◽  
Author(s):  
Soo-Wang Hyun ◽  
Junghyun Kim ◽  
Bongkyun Park ◽  
Kyuhyung Jo ◽  
Tae Lee ◽  
...  
Keyword(s):  
Particulate Matter ◽  
Epithelial Cells ◽  
Ocular Surface ◽  
Keratoconjunctivitis Sicca ◽  
Bioactive Compound ◽  
Protective Effects ◽  
Apricot Kernel ◽  
Urban Particulate Matter ◽  
Tnf Α ◽  
Conjunctival Epithelial Cells

Exposure to particulate matter is a risk factor for various ocular surface diseases, including keratoconjunctivitis sicca (KCS). In this study, we investigated the protective effects of apricot kernel extract (AKE) and its bioactive compound, amygdalin, on KCS induced by exposure to urban particulate matter (UPM). In the in vivo experiments, eye drops containing 0.5 mg/mL AKE (AKE-0.5) or 1 mg/mL AKE (AKE-1) were administered directly into the eyes of female rats after UPM exposure. Additionally, the effect of AKE and amygdalin on matrix metalloproteinases (MMPs) activity and the expressions of inflammatory factors, including tumor necrosis factor (TNF)-α and interleukin (IL)-6, was investigated in conjunctival epithelial cells in vitro. Topical administration of AKE-1 attenuated UPM exposure-induced reduction of tear secretion. Both AKE-0.5 and AKE-1 inhibited UPM exposure-induced corneal epithelial damage and irregularity. AKE also protected against UPM exposure-induced disruption of the mucin-4 layer on the ocular surface. In addition, AKE and amygdalin prevented UPM-induced activation of MMPs and upregulation of TNF-α and IL-6 in conjunctival epithelial cells. Therefore, AKE may have protective effects against UPM exposure-induced KCS via the inhibition of MMPs and inflammation. The pharmacological activities of AKE may be in part due to its bioactive compound, amygdalin.

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