scholarly journals Investigation of Cytotoxic Effects of Recombinant Human Interferon Lambda-1 and Its Pegylated Form on Human Conjunctival Epithelial Cells

2021 ◽  
Vol 9 (4) ◽  
pp. 200-208
Author(s):  
N. A. Kikhtenko ◽  
N. A. Bondarenko ◽  
N. P. Bgatova ◽  
L. A. Oleynik ◽  
O. V. Poveshchenko ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Proliferative Activity ◽  
Cytotoxic Effect ◽  
Ultrastructural Changes ◽  
Human Interferon ◽  
Interferon Lambda ◽  
Study Results ◽  
Cell Proliferative Activity ◽  
Pegylated Form

Currently, there are no efficacious, all-purpose antiviral medicines for the treatment of ocular surface infections caused by viruses. At the same time, type III interferons demonstrate high potency for histological barriers, such as the conjunctiva. Modification of protein molecules in native products can significantly improve their pharmacodynamic properties. Thus, it seems reasonable to develop antiviral medicines based on interferon lambda (IFN-λ1) and its pegylated form (PEG IFN-λ1).The aim of the study was to evaluate the in vitro cytotoxic effect of recombinant human IFN-λ1 and its pegylated form on Chang conjunctiva clone 1-5c-4 human conjunctival cells.Materials and methods: PEG IFN-λ1 was obtained by the electron beam immobilisation method. A normal human conjunctival cell line Chang conjunctiva clone 1-5c-4 was used for cell cultivation. The MTT test was used to assess the cytotoxic effect. Cell proliferative activity was studied by measuring microelectrode impedance. Ultrastructural changes were assessed by electron microscopy. Statistical processing was performed using the Statistica 10.0 software package.Results: IFN-λ1 (37 μg/mL) and PEG IFN-λ1 (42 μg/mL) had no significant cytotoxic effect on the human conjunctiva cell culture and the cell proliferative activity. The analysis of ultrastructural changes demonstrated that IFN-λ1 activated metabolic processes in the cells, and PEG IFN-λ1 promoted differentiation and keratinisation of epithelial cells and led to modification of the cell membrane. A ten-fold increase in IFN-λ1 and PEG IFN-λ1 concentration (to 370 μg/mL and 420 μg/mL, respectively) reduced the cell viability by 15–20% as compared to the intact control.Conclusions: the study results demonstrated that IFN-λ1 and PEG IFN-λ1 could be used as active pharmaceutical ingredients in the development of medicines for the treatment of conjunctival viral infections.

scholarly journals The effect of TGF-β1 and Smad7 gene transfer on the phenotypic changes of rat alveolar epithelial cells

2007 ◽  
Vol 12 (3) ◽  
Author(s):  
Guo-Ping Xu ◽  
Qing-Quan Li ◽  
Xi-Xi Cao ◽  
Qi Chen ◽  
Zhong-Hua Zhao ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Epithelial Cells ◽  
Transforming Growth Factor ◽  
Alveolar Epithelial Cells ◽  
Ultrastructural Changes ◽  
Type Ii ◽  
Mesenchymal Transition ◽  
Alveolar Epithelial ◽  
Tgf Β1

AbstractThe aim of this study was to investigate whether transforming growth factor-β1 (TGF-β1) could induce alveolar epithelial-mesenchymal transition (EMT) in vitro, and whether Smad7 gene transfer could block this transition. We also aimed to elucidate the possible mechanisms of these processes. The Smad7 gene was transfected to the rat type II alveolar epithelial cell line (RLE-6TN). Expression of the EMT-associated markers was assayed by Western Blot and Real-time PCR. Morphological alterations were examined via phase-contrast microscope and fluorescence microscope, while ultrastructural changes were examined via electron microscope. TGF-β1 treatment induced a fibrotic phenotype of RLE-6TN with increased expression of fibronectin (FN), α-smooth muscle actin (α-SMA) and vimentin, and decreased expression of E-cadherin (E-cad) and cytokeratin19 (CK19). After transfecting the RLE-6TN with the Smad7 gene, the expression of the mesenchymal markers was downregulated while that of the epithelial markers was upregulated. TGF-β1 treatment for 48 h resulted in the separation of RLE-6TN from one another and a change into elongated, myofibroblast-like cells. After the RLE-6TN had been transfected with the Smad7 gene, TGF-β1 treatment had no effect on the morphology of the RLE-6TN. TGF-β1 treatment for 48 h resulted in an abundant expression of α-SMA in the RLE-6TN. If the RLE-6TN were transfected with the Smad7 gene, TGF-β1 treatment for 48 h could only induce a low level of α-SMA expression. Furthermore, TGF-β1 treatment for 12 h resulted in the degeneration and swelling of the osmiophilic multilamellar bodies, which were the markers of type II alveolar epithelial cells. TGF-β1 can induce alveolar epithelialmesenchymal transition in vitro, which is dependent on the Smads signaling pathway to a certain extent. Overexpression of the Smad7 gene can partially block this process

scholarly journals Epidermal growth factor reduces autophagy in intestinal epithelium and in the rat model of necrotizing enterocolitis

2010 ◽  
Vol 299 (3) ◽  
pp. G614-G622 ◽  
Author(s):  
Andrew A. Maynard ◽  
Katerina Dvorak ◽  
Ludmila Khailova ◽  
Holly Dobrenen ◽  
Kelly M. Arganbright ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Epithelial Cells ◽  
Necrotizing Enterocolitis ◽  
Intestinal Epithelium ◽  
Beclin 1 ◽  
Ultrastructural Changes ◽  
Epidermal Growth

Necrotizing enterocolitis (NEC) is a devastating intestinal disease of premature infants. Epidermal growth factor (EGF) is one of the most promising candidates in NEC prophylaxis. Autophagy regulates cell homeostasis, but uncontrolled activation of autophagy may lead to cellular injury. The aim was to evaluate the effects of EGF on intestinal autophagy in epithelial cells and in the rat NEC model and measure autophagy in NEC patients. Intestinal epithelial cells (IEC-6) and the rat NEC model were used to study the effect of EGF on intestinal autophagy. Protein levels of Beclin 1 and LC3II were measured in the intestinal epithelium in both in vivo and in vitro models. Ultrastructural changes in intestinal epithelium were studied by electron microscopy. Expression of Beclin 1, LC3II, and p62 protein was evaluated in biopsies from NEC patients. Autophagy was induced in IEC-6 cells and inhibited by adding EGF into the culture. In the rat NEC model, EGF treatment of NEC reduced expression of Beclin 1 and LC3II in ileal epithelium. Morphologically, typical signs of autophagy were observed in the epithelium of the NEC group, but not in the EGF group. A strong signal for Beclin 1 and LC3II was detected in the intestine from patients with NEC. Autophagy is activated in the intestinal epithelium of NEC patients and in the ileum of NEC rats. Supplementation of EGF blocks intestinal autophagy in both in vivo and in vitro conditions. Results from this study indicate that EGF-mediated protection against NEC injury is associated with regulation of intestinal autophagy.

scholarly journals Epithelium of mouse yolk sac placenta lacks H-2 complex alloantigens.

1980 ◽  
Vol 152 (4) ◽  
pp. 945-955 ◽  
Author(s):  
E L Parr ◽  
R V Blanden ◽  
R S Tulsi
Keyword(s):  
Epithelial Cells ◽  
Cytotoxic Effect ◽  
Apical Membrane ◽  
Vascular Endothelial Cells ◽  
Yolk Sac ◽  
Mhc Antigens ◽  
Mhc Molecules ◽  
Apical Membranes ◽  
Yolk Sac Placenta

The only fetal cell membrane exposed to the mother in the mouse yolk sac placenta is the apical membrane of the endodermal epithelial cells. In yolk sac preparations in vitro, this apical membrane was exposed to reagents or cells in the incubation medium. By using several techniques we were not able to detect fetal major histocompatibility complex (MHC) antigens in this membrane. Immunoferritin labeling with and without prefixation and after neurominidase and trypsin digestion indicated that the apical membrane could contain no more than approximately 1% of the H-2 complex antigens that were present on peritoneal macrophages. Incubation of yolk sac preparations in anti-H-2 complex antiserum and complement had no cytotoxic effect on the endodermal epithelium, nor did incubation in an excess of alloreactive lymphocytes. Dissociated preparations of prefixed yolk sac contained endodermal epithelial cells and vascular endothelial cells whose entire surface membranes were exposed to the medium. H-2-complex antigens were not detected by immunoferritin labeling in either the apical or the laterobasal membrane of the yolk sac endoderm, but they were present in low density on the vascular endothelium. Also, incubation of unfixed, dissociated cells in anti-H-2-complex serum and complement had no detectable cytotoxic effect on endodermal epithelial cells. These observations indicate that H-2 antigens are sparse or absent in both the apical and laterobasal membranes of endodermal epithelial cells. The deficiency of MHC antigens in the apical membrane may account for the failure of sensitized females to reject the yolk sac, whereas the composition of the laterobasal membrane is probably less important to maternal-fetal relations. The present observations are consistent with labeling studies of adult-lining epithelial cells, which indicate that self-marker MHC molecules are absent from the apical membranes oriented toward the outside world and variably expressed in the laterobasal self-side membranes. It is suggested that the corresponding exclusion of fetal self-marker molecules from the apical membranes of some kinds of placental epithelia would deprive the mother of target sites for an alloimmune reaction at the maternal-fetal interface.

scholarly journals Interferon Lambda Delays the Emergence of Influenza Virus Resistance to Oseltamivir

2021 ◽  
Vol 9 (6) ◽  
pp. 1196
Author(s):  
Chiara Medaglia ◽  
Arnaud Charles-Antoine Zwygart ◽  
Paulo Jacob Silva ◽  
Samuel Constant ◽  
Song Huang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Neuraminidase Inhibitor ◽  
Influenza Viruses ◽  
Human Interferon ◽  
Infection Model ◽  
Single Mutation ◽  
Resistance Mutations ◽  
Species Barrier ◽  
Interferon Lambda

Influenza viruses are a leading cause of morbidity and mortality worldwide. These air-borne pathogens are able to cross the species barrier, leading to regular seasonal epidemics and sporadic pandemics. Influenza viruses also possess a high genetic variability, which allows for the acquisition of resistance mutations to antivirals. Combination therapies with two or more drugs targeting different mechanisms of viral replication have been considered an advantageous option to not only enhance the effectiveness of the individual treatments, but also reduce the likelihood of resistance emergence. Using an in vitro infection model, we assessed the barrier to viral resistance of a combination therapy with the neuraminidase inhibitor oseltamivir and human interferon lambda against the pandemic H1N1 A/Netherlands/602/2009 (H1N1pdm09) virus. We serially passaged the virus in a cell line derived from human bronchial epithelial cells in the presence or absence of increasing concentrations of oseltamivir alone or oseltamivir plus interferon lambda. While the treatment with oseltamivir alone quickly induced the emergence of antiviral resistance through a single mutation in the neuraminidase gene, the co-administration of interferon lambda delayed the emergence of drug-resistant influenza virus variants. Our results suggest a possible clinical application of interferon lambda in combination with oseltamivir to treat influenza.

scholarly journals P 125 Lack of cytotoxic effect of ganciclovir and acyclovir on rabbit corneal epithelial cells in vitro

1995 ◽  
Vol 35 ◽  
pp. S173
Author(s):  
E. Khosravi ◽  
P.P. Elena ◽  
P. Goldschmidt ◽  
C. Baudoin ◽  
J. Luyckx
Keyword(s):  
Epithelial Cells ◽  
Cytotoxic Effect ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial

scholarly journals Influence of different concentrations of 6-benzylaminopurine and thidiazuron on the proliferative activity of apple varieties in in vitro culture

2021 ◽  
Vol 36 ◽  
pp. 03012
Author(s):  
L.V. Tashmatova ◽  
O.V. Matsneva ◽  
T.M. Khromova ◽  
V.V. Shakhov
Keyword(s):  
In Vitro Culture ◽  
Apical Dominance ◽  
Proliferative Activity ◽  
Essential Role ◽  
Cytokinin Activity ◽  
Multiplication Factor ◽  
Apple Varieties ◽  
Clonal Micropropagation ◽  
Study Results

An essential role in the process of clonal micropropagation is played by cytokinins, with the help of which apical dominance is removed and the formation of additional shoots and buds is activated. The article presents the study results of microshoots’ proliferative activity of apple varieties on media with 6-benzylaminopurine at a concentration of 1.0 mg/l, 2.0 mg/l and thidiazuron at a concentration of 0.1 mg/l, 0.2 mg/l and 0.3 mg/l. The objects of research were apple varieties of the VNIISPK selection - Bolotovskoe, Imrus, Veteran, Kandil orlovsky, Girlyanda, Priokskoe. The aim of the research was to study the effect of substances with cytokinin activity on the morphogenesis of apple varieties. As a result of the research, it was revealed that both cytokinins and varietal characteristics influence the multiplication factor. The highest multiplication factor was observed when using 6-BAP. Thidiazuron reduced the proliferation rate. When using 6-BAP, Bolotovskoe, Girlyanda and Priokskoe varieties had the highest multiplication factor among the varieties. When using TDZ - varieties Bolotovskoe and Kandil Orlovsky.

scholarly journals Response of cultured normal canine mammary epithelial cells to deracoxib—doxorubicin combination

2017 ◽  
Vol 65 (3) ◽  
pp. 366-381
Author(s):  
Tulay Bakirel ◽  
Fulya Ustun Alkan ◽  
Oya Ustuner ◽  
Suzan Çinar ◽  
Ceren Anlas ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Cytotoxic Action ◽  
Current Standard ◽  
Griess Reaction ◽  
Study Results

Currently, there is a growing interest in combining anticancer drugs with the aim to improve outcome in patients suffering from tumours and reduce the long-term toxicity associated with the current standard of treatment. In this study, we evaluated the possible role of deracoxib against the toxicity of doxorubicin on normal canine mammary epithelial cells. The effect of deracoxib and doxorubicin combination on cell viability was determined by MTT assay. Apoptosis was characterised by flow cytometry. Cell nitrite concentrations were measured with the Griess reaction. Deracoxib (50 and 100 μM) treatment decreased the cytotoxic action of doxorubicin at 0.9 μM in the cells, from 33.63% to 13.4% and 25.82%, respectively. Our results also showed that the reverse effect of deracoxib on doxorubicin-induced cytotoxic activity in the cells was associated with a marked (3.04- to 3.57-fold) decrease in apoptosis. In additional studies identifying the mechanism of the observed effect, deracoxib exhibited an activity to prevent doxorubicin-mediated overproduction of nitric oxide in the cells. Our in vitro study results indicate that deracoxib (50 and 100 μM) can be beneficial in protecting normal cells from the toxic effect of doxorubicin in conjunction with apoptosis by the modulation of nitric oxide production.

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