Cargando…
Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis
A defective-interfering (DI) RNA of mouse hepatitis virus (MHV) was developed as a vector for expressing MHV hemagglutinin/esterase (HE) protein. The virus containing an expressed HE protein (A59-DE-HE) was generated by infecting cells with MHV-A59, which does not express HE, and transfecting thein...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academic Press.
1998
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131006/ https://www.ncbi.nlm.nih.gov/pubmed/9501044 http://dx.doi.org/10.1006/viro.1997.8993 |
_version_ | 1783517139250970624 |
---|---|
author | Zhang, Xuming Hinton, David R. Park, Sungmin Parra, Beatriz Liao, Ching-Len Lai, Michael M.C. Stohlman, Stephen A. |
author_facet | Zhang, Xuming Hinton, David R. Park, Sungmin Parra, Beatriz Liao, Ching-Len Lai, Michael M.C. Stohlman, Stephen A. |
author_sort | Zhang, Xuming |
collection | PubMed |
description | A defective-interfering (DI) RNA of mouse hepatitis virus (MHV) was developed as a vector for expressing MHV hemagglutinin/esterase (HE) protein. The virus containing an expressed HE protein (A59-DE-HE) was generated by infecting cells with MHV-A59, which does not express HE, and transfecting thein vitro-transcribed DI RNA containing the HE gene. A similar virus (A59-DE-CAT) expressing the chloramphenicol acetyltransferase (CAT) was used as a control. These viruses were inoculated intracerebrally into mice, and the role of the HE protein in viral pathogenesis was evaluated. Results showed that all mice infected with parental A59 or A59-DE-CAT succumbed to infection by 9 days postinfection (p.i.), demonstrating that inclusion of the DI did not by itself alter pathogenesis. In contrast, 60% of mice infected with A59-DE-HE survived infection. HE- or CAT-specific subgenomic mRNAs were detected in the brains at days 1 and 2 p.i. but not later, indicating that the genes in the DI vector were expressed only in the early stage of viral infection. No significant difference in virus titer or viral antigen expression in brains was observed between A59-DE-HE- and A59-DE-CAT-infected mice, suggesting that virus replication in brain was not affected by the expression of HE. However, at day 3 p.i. there was a slight increase in the extent of inflammatory cell infiltration in the brains of the A59-DE-HE-infected mice. Surprisingly, virus titers in the livers of A59-DE-HE-infected mice were 3 log(10)lower than that of the A59-DE-CAT-infected mice at day 6 p.i. Also, substantially less necrosis and viral antigen were detected in the livers of the A59-DE-HE-infected mice. This may account for the reduced mortality of these mice. The possible contribution of the host immune system to this difference in pathogenesis was analyzed by comparing the expression of four cytokines. Results showed that both tumor necrosis factor-α and interleukin-6 mRNAs increased in the brains of the A59-DE-HE-infected mice at day 2 p.i., whereas interferon-γ and interleukin-1α mRNAs were similar between A59-DE-HE- and A59-DE-CAT-infected mice. These data suggest that the transient expression of HE protein enhances an early innate immune response, possibly contributing to the eventual clearance of virus from the liver. This study indicates the feasibility of the DI expression system for studying roles of viral proteins during MHV infection. |
format | Online Article Text |
id | pubmed-7131006 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1998 |
publisher | Academic Press. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71310062020-04-08 Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis Zhang, Xuming Hinton, David R. Park, Sungmin Parra, Beatriz Liao, Ching-Len Lai, Michael M.C. Stohlman, Stephen A. Virology Regular Article A defective-interfering (DI) RNA of mouse hepatitis virus (MHV) was developed as a vector for expressing MHV hemagglutinin/esterase (HE) protein. The virus containing an expressed HE protein (A59-DE-HE) was generated by infecting cells with MHV-A59, which does not express HE, and transfecting thein vitro-transcribed DI RNA containing the HE gene. A similar virus (A59-DE-CAT) expressing the chloramphenicol acetyltransferase (CAT) was used as a control. These viruses were inoculated intracerebrally into mice, and the role of the HE protein in viral pathogenesis was evaluated. Results showed that all mice infected with parental A59 or A59-DE-CAT succumbed to infection by 9 days postinfection (p.i.), demonstrating that inclusion of the DI did not by itself alter pathogenesis. In contrast, 60% of mice infected with A59-DE-HE survived infection. HE- or CAT-specific subgenomic mRNAs were detected in the brains at days 1 and 2 p.i. but not later, indicating that the genes in the DI vector were expressed only in the early stage of viral infection. No significant difference in virus titer or viral antigen expression in brains was observed between A59-DE-HE- and A59-DE-CAT-infected mice, suggesting that virus replication in brain was not affected by the expression of HE. However, at day 3 p.i. there was a slight increase in the extent of inflammatory cell infiltration in the brains of the A59-DE-HE-infected mice. Surprisingly, virus titers in the livers of A59-DE-HE-infected mice were 3 log(10)lower than that of the A59-DE-CAT-infected mice at day 6 p.i. Also, substantially less necrosis and viral antigen were detected in the livers of the A59-DE-HE-infected mice. This may account for the reduced mortality of these mice. The possible contribution of the host immune system to this difference in pathogenesis was analyzed by comparing the expression of four cytokines. Results showed that both tumor necrosis factor-α and interleukin-6 mRNAs increased in the brains of the A59-DE-HE-infected mice at day 2 p.i., whereas interferon-γ and interleukin-1α mRNAs were similar between A59-DE-HE- and A59-DE-CAT-infected mice. These data suggest that the transient expression of HE protein enhances an early innate immune response, possibly contributing to the eventual clearance of virus from the liver. This study indicates the feasibility of the DI expression system for studying roles of viral proteins during MHV infection. Academic Press. 1998-03-01 2002-05-25 /pmc/articles/PMC7131006/ /pubmed/9501044 http://dx.doi.org/10.1006/viro.1997.8993 Text en Copyright © 1998 Academic Press. All rights reserved. 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 | Regular Article Zhang, Xuming Hinton, David R. Park, Sungmin Parra, Beatriz Liao, Ching-Len Lai, Michael M.C. Stohlman, Stephen A. Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title | Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title_full | Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title_fullStr | Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title_full_unstemmed | Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title_short | Expression of Hemagglutinin/Esterase by a Mouse Hepatitis Virus Coronavirus Defective–Interfering RNA Alters Viral Pathogenesis |
title_sort | expression of hemagglutinin/esterase by a mouse hepatitis virus coronavirus defective–interfering rna alters viral pathogenesis |
topic | Regular Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131006/ https://www.ncbi.nlm.nih.gov/pubmed/9501044 http://dx.doi.org/10.1006/viro.1997.8993 |
work_keys_str_mv | AT zhangxuming expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT hintondavidr expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT parksungmin expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT parrabeatriz expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT liaochinglen expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT laimichaelmc expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis AT stohlmanstephena expressionofhemagglutininesterasebyamousehepatitisviruscoronavirusdefectiveinterferingrnaaltersviralpathogenesis |