Article

Susceptibility of human and rat neural cell lines to infection by SARS-coronavirus.

Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
Biochemical and Biophysical Research Communications (impact factor: 2.48). 08/2005; 334(1):79-85. DOI:10.1016/j.bbrc.2005.06.061 pp.79-85
Source: PubMed

ABSTRACT Pathological characterization of autopsied tissues from patients with SARS revealed severe damage in restricted tissues, such as lung, with no apparent cell damage in other tissues, such as intestine and brain. Here, we examined the susceptibility of neural cell lines of human (OL) and rat (C6) origins to SARS-associated coronavirus. Both of the neural cell lines showed no apparent cytopathic effects (CPE) by infection but produced virus with infectivity of 10(2-5) per ml, in sharp contrast to the production by infected Vero E6 cells of >10(9) per ml that showed a lytic infection with characteristic rounding CPE. Interestingly, the infection of intestinal cell line CaCo-2 also induced no apparent CPE, with production of the virus at a slightly lower level as that of the Vero E6 cell culture. Notably, the cellular receptor for the virus, angiotensin-converting enzyme 2 was expressed at similar levels on Vero E6 and CaCo-2 cells, but at undetectable levels on OL and C6 cells.

0 0
 · 
0 Bookmarks
 · 
12 Views
  • Source
    Article: Plaque assay for human coronavirus NL63 using human colon carcinoma cells.
    Petra Herzog, Christian Drosten, Marcel A Müller
    [show abstract] [hide abstract]
    ABSTRACT: Coronaviruses cause a broad range of diseases in animals and humans. Human coronavirus (hCoV) NL63 is associated with up to 10% of common colds. Viral plaque assays enable the characterization of virus infectivity and allow for purifying virus stock solutions. They are essential for drug screening. Hitherto used cell cultures for hCoV-NL63 show low levels of virus replication and weak and diffuse cytopathogenic effects. It has not yet been possible to establish practicable plaque assays for this important human pathogen. 12 different cell cultures were tested for susceptibility to hCoV-NL63 infection. Human colon carcinoma cells (CaCo-2) replicated virus more than 100 fold more efficiently than commonly used African green monkey kidney cells (LLC-MK2). CaCo-2 cells showed cytopathogenic effects 4 days post infection. Avicel, agarose and carboxymethyl-cellulose overlays proved suitable for plaque assays. Best results were achieved with Avicel, which produced large and clear plaques from the 4th day of infection. The utility of plaque assays with agrose overlay was demonstrated for purifying virus, thereby increasing viral infectivity by 1 log 10 PFU/mL. CaCo-2 cells support hCoV-NL63 better than LLC-MK2 cells and enable cytopathogenic plaque assays. Avicel overlay is favourable for plaque quantification, and agarose overlay is preferred for plaque purification. HCoV-NL63 virus stock of increased infectivity will be beneficial in antiviral screening, animal modelling of disease, and other experimental tasks.
    Virology Journal 12/2008; 5:138. · 2.34 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

angiotensin-converting enzyme 2
 
apparent cell damage
 
apparent CPE
 
autopsied tissues
 
C6 cells
 
CaCo-2 cells
 
cellular receptor
 
Interestingly
 
intestinal cell line CaCo-2
 
lytic infection
 
neural cell lines
 
Pathological characterization
 
SARS-associated coronavirus
 
severe damage
 
sharp contrast
 
similar levels
 
undetectable levels
 
Vero E6
 
Vero E6 cell culture
 
Vero E6 cells
 

Makiko Yamashita