scholarly journals ENUMERATION OF CELL-INFECTING PARTICLES OF NEWCASTLE DISEASE VIRUS BY THE FLUORESCENT ANTIBODY TECHNIQUE

1961 ◽  
Vol 113 (2) ◽  
pp. 301-316 ◽  
Author(s):  
E. Frederick Wheelock ◽  
Igor Tamm
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Hela Cells ◽  
Newcastle Disease ◽  
Fluorescent Antibody ◽  
Cell Counting ◽  
Virus Concentration ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Counting Procedure

A procedure has been developed for the determination of the concentration of infective Newcastle disease virus (NDV) based on the enumeration of singly infected and distributed HeLa cells which are visualized by staining with fluorescent antibody. Infective virus assayed by the fluorescent cell-counting procedure is expressed in terms of cell-infecting units (CIU). Adsorption of NDV to HeLa cell monolayers reached a plateau 1 to 1.5 hours after inoculation of coverslip cultures, and 12 per cent of the infective particles inoculated failed to adsorb. The half-life of NDV in protein-free Eagle's medium at 37°C. was 2.1 hours. There was a linear relationship between virus concentration and the number of infected cells. The coefficient of variation of the mean of replicate determinations of infective NDV was 8.2 per cent. The distribution of single infected HeLa cells in the monolayer corresponded to the Poisson distribution. With NDV the cell-infecting unit (CIU) determined in HeLa cells is equivalent to the plaque-forming unit in chick embryo cells and the egg infective dose. In experiments on the mechanism of dissemination of NDV in monolayer cultures of HeLa cells, NDV was found to spread from cell to cell through the extracellular milieu.

scholarly journals Newcastle disease virus may enter cells by caveolae-mediated endocytosis

2007 ◽  
Vol 88 (2) ◽  
pp. 559-569 ◽  
Author(s):  
Celia Cantín ◽  
Javier Holguera ◽  
Laura Ferreira ◽  
Enrique Villar ◽  
Isabel Muñoz-Barroso
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Inhibitory Effect ◽  
Endocytic Pathway ◽  
Host Cells ◽  
Virus Concentration ◽  
Double Labelling ◽  
Virus Adsorption ◽  
Labelling Immunofluorescence

The entry into cells of Newcastle disease virus (NDV), a prototype member of the paramyxoviruses, is believed to occur by direct fusion at the plasma membrane through a pH-independent mechanism. In addition, NDV may enter host cells by an endocytic pathway. Treatment of cells with drugs that block caveolae-dependent endocytosis reduced NDV fusion and infectivity, the degree of inhibition being dependent on virus concentration. The inhibitory effect was reduced greatly when drugs were added after virus adsorption. Cells treated with methyl β-cyclodextrin, a drug that sequesters cholesterol from membranes, reduced the extent of fusion, infectivity and virus–cell binding; this indicates that cholesterol plays a role in NDV entry. Double-labelling immunofluorescence assays performed with anti-NDV monoclonal antibodies and antibodies against the early endosome marker EEA1 revealed the localization of the virus in these intracellular structures. Using fluorescence microscopy, it was found that cell–cell fusion was enhanced at low pH. It is concluded that NDV may infect cells through a caveolae-dependent endocytic pathway, suggesting that this pathway could be an alternative route for virus entry into cells.

Inactivation of Aerosolized Newcastle Disease Virus with Non-thermal Plasma

2020 ◽  
Vol 36 (1) ◽  
pp. 55-60
Author(s):  
Charles Schiappacasse ◽  
Peng Peng ◽  
Nan Zhou ◽  
Xiaoying Liu ◽  
Jie Zhai ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Thermal Plasma ◽  
Limit Of Detection ◽  
Virus Concentration ◽  
List Type ◽  
Airborne Transmission ◽  
Flow Rates ◽  
Non Thermal Plasma

Abstract.HighlightsNon-thermal plasma treatment completely inactivated Newcastle disease virus (NDV).Higher airborne virus concentrations at lower humidity levels & higher flow rates.Nebulization (20 min) did not inactivate NDV .Abstract.The airborne transmission of poultry viruses, such as Newcastle disease virus, is major health and economic concern. The poultry industry currently lacks a cost-effective solution to prevent airborne transmission. The present study explored non-thermal plasma’s ability to inactivate poultry viruses, by challenging a laboratory-scale non-thermal plasma reactor with aerosolized Newcastle disease virus at increasing flow rates (i.e., decreased direct treatment time). Viruses were inactivated below the green fluorescent protein (GFP) focus-forming units per mL (GFU/mL) limit of detection at the flow rates 18, 23, and 28 liters per minute (LPM). However, this study did not differentiate between inactivation effects caused by direct NTP treatment and indirect NTP treatment (viruses exposed to ozone after collection on gelatin membrane filters). A strong relationship (R2 =.99) was observed between decreasing relative humidity and increasing airborne virus concentrations. Twenty minutes of nebulization did not significantly change liquid virus concentration in the nebulizer. Keywords: Aerosol, Airborne, Newcastle disease, Non-thermal, Pathogen, Plasma, Virus.

scholarly journals Exosomes Carry microRNAs into Neighboring Cells to Promote Diffusive Infection of Newcastle Disease Virus

Viruses ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 527 ◽  
Author(s):  
Changluan Zhou ◽  
Lei Tan ◽  
Yingjie Sun ◽  
Xusheng Qiu ◽  
Chunchun Meng ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Tumor Cells ◽  
Disease Virus ◽  
Hela Cells ◽  
Newcastle Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Bioactive Substances ◽  
Cytopathic Effects

Newcastle disease virus (NDV), an avian paramyxovirus, was shown to prefer to replicate in tumor cells instead of normal cells; however, this mechanism has not been fully elucidated. Exosomes play a crucial role in intercellular communication due to the bioactive substances they carry. Several studies have shown that exosomes are involved in virus infections. However, the effect that exosomes have on NDV-infected tumor cells is not known. In this study, we focus on the role of exosomes secreted by NDV-infected HeLa cells in promoting NDV replication. Three miRNA candidates (miR-1273f, miR-1184, and miR-198) embraced by exosomes were associated with enhancing NDV-induced cytopathic effects on HeLa cells. Furthermore, luciferase assays, RT-qPCR, and enzyme-linked immunosorbent assay (ELISA) all demonstrated that these miRNAs could suppress interferon (IFN)-β gene expression. Enhanced NDV replication in HeLa cells was identified by Western blot and plaque assays. Based on these results, we speculate that NDV employed exosomes entry into neighboring cells, which carry miRNAs, resulting in inhibition of the IFN pathway and promotion of viral infection. To our knowledge, this is the first report on the involvement of NDV-employed exosomes in tumor cells, and as such, it provides new insights into the development of anti-tumor therapies.

HN protein of Newcastle disease virus sensitizes HeLa cells to TNF-α-induced apoptosis by downregulating NF-κB expression

2016 ◽  
Vol 161 (9) ◽  
pp. 2395-2405 ◽  
Author(s):  
R. S. Rajmani ◽  
Shishir Kumar Gupta ◽  
Prafull Kumar Singh ◽  
Ravi Kumar Gandham ◽  
A. P. Sahoo ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Hela Cells ◽  
Newcastle Disease ◽  
Tnf Α ◽  
Induced Apoptosis
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