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Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties
Twenty 5-fluorouracil-induced temperature-sensitive (ts) mutants of mouse hepatitis virus strain A59 were isolated from 1284 virus clones. Mutants were preselected on the basis of their inability to induce syncytia in infected cells at the restrictive temperature (40°) vs the permissive temperature...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Published by Elsevier Inc.
1983
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126919/ https://www.ncbi.nlm.nih.gov/pubmed/6301146 http://dx.doi.org/10.1016/0042-6822(83)90211-8 |
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author | Koolen, Marck J.M. Osterhaus, Albert D.M.E. Van Steenis, Gijsbert Horzinek, Marian C. van der Zeijst, Bernard A.M. |
author_facet | Koolen, Marck J.M. Osterhaus, Albert D.M.E. Van Steenis, Gijsbert Horzinek, Marian C. van der Zeijst, Bernard A.M. |
author_sort | Koolen, Marck J.M. |
collection | PubMed |
description | Twenty 5-fluorouracil-induced temperature-sensitive (ts) mutants of mouse hepatitis virus strain A59 were isolated from 1284 virus clones. Mutants were preselected on the basis of their inability to induce syncytia in infected cells at the restrictive temperature (40°) vs the permissive temperature (31°). Of these mutants, only those with a relative plating efficiency 40°31° of 3 × 10(−3) or smaller were kept. Virus yields at 40° compared to 37° and 31° (leakiness) were determined. Most mutants (16) were RNA(−), i.e., unable to synthesize virus-specific RNA at the restrictive temperature. The other four were RNA(+). No qualitative differences were detected in the virus-specific RNAs in cells infected with RNA(+) ts-mutants, both at 31° and 40°. Virus-specific proteins present in cells infected with is-171 (RNA(−)) and the RNA(+)-mutants (ts-43, is-201, is-209, and is-279) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitates. No qualitative differences in the pattern of virus-specific cellular proteins were detected among the mutants except for an additional polypeptide of about 46,000 daltons in ts-209-infected cells. Finally, the neuropathogenic properties of eight of the mutants were investigated. Whereas 10(2) PFU of wild-type virus injected intracerebrally killed 50 to 100% of 4-week-old Balc/c mice within 1 week, the mutants were highly attenuated. A dose of 10(5) PFU lead to no or transient disease. However, 4 weeks after infection with ts 342, is-43, or is-201 obvious histological changes were observed in brain and spinal cord of clinically healthy mice. |
format | Online Article Text |
id | pubmed-7126919 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1983 |
publisher | Published by Elsevier Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71269192020-04-08 Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties Koolen, Marck J.M. Osterhaus, Albert D.M.E. Van Steenis, Gijsbert Horzinek, Marian C. van der Zeijst, Bernard A.M. Virology Article Twenty 5-fluorouracil-induced temperature-sensitive (ts) mutants of mouse hepatitis virus strain A59 were isolated from 1284 virus clones. Mutants were preselected on the basis of their inability to induce syncytia in infected cells at the restrictive temperature (40°) vs the permissive temperature (31°). Of these mutants, only those with a relative plating efficiency 40°31° of 3 × 10(−3) or smaller were kept. Virus yields at 40° compared to 37° and 31° (leakiness) were determined. Most mutants (16) were RNA(−), i.e., unable to synthesize virus-specific RNA at the restrictive temperature. The other four were RNA(+). No qualitative differences were detected in the virus-specific RNAs in cells infected with RNA(+) ts-mutants, both at 31° and 40°. Virus-specific proteins present in cells infected with is-171 (RNA(−)) and the RNA(+)-mutants (ts-43, is-201, is-209, and is-279) were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitates. No qualitative differences in the pattern of virus-specific cellular proteins were detected among the mutants except for an additional polypeptide of about 46,000 daltons in ts-209-infected cells. Finally, the neuropathogenic properties of eight of the mutants were investigated. Whereas 10(2) PFU of wild-type virus injected intracerebrally killed 50 to 100% of 4-week-old Balc/c mice within 1 week, the mutants were highly attenuated. A dose of 10(5) PFU lead to no or transient disease. However, 4 weeks after infection with ts 342, is-43, or is-201 obvious histological changes were observed in brain and spinal cord of clinically healthy mice. Published by Elsevier Inc. 1983-03 2004-02-06 /pmc/articles/PMC7126919/ /pubmed/6301146 http://dx.doi.org/10.1016/0042-6822(83)90211-8 Text en Copyright © 1983 Published by Elsevier Inc. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Koolen, Marck J.M. Osterhaus, Albert D.M.E. Van Steenis, Gijsbert Horzinek, Marian C. van der Zeijst, Bernard A.M. Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title | Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title_full | Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title_fullStr | Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title_full_unstemmed | Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title_short | Temperature-sensitive mutants of mouse hepatitis virus strain A59: Isolation, characterization and neuropathogenic properties |
title_sort | temperature-sensitive mutants of mouse hepatitis virus strain a59: isolation, characterization and neuropathogenic properties |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7126919/ https://www.ncbi.nlm.nih.gov/pubmed/6301146 http://dx.doi.org/10.1016/0042-6822(83)90211-8 |
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