Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems

<p>Abstract</p> <p>Background</p> <p>Highly pathogenic Avian Influenza (HPAI) virus is able to infect many hosts and the virus replicates in high levels in the respiratory tract inducing severe lung lesions. The pathogenesis of the disease is actually the outcome of the...

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Main Authors: Noor Suriani M, Mohamed Maizan, Omar Abdul R, Hassan Sharifah S, Balasubramaniam Vinod RMT, Mohamed Ramlan, Othman Iekhsan
Format: Article
Language:English
Published: BMC 2011-04-01
Series:Virology Journal
Subjects:
Online Access:http://www.virologyj.com/content/8/1/196
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author Noor Suriani M
Mohamed Maizan
Omar Abdul R
Hassan Sharifah S
Balasubramaniam Vinod RMT
Mohamed Ramlan
Othman Iekhsan
author_facet Noor Suriani M
Mohamed Maizan
Omar Abdul R
Hassan Sharifah S
Balasubramaniam Vinod RMT
Mohamed Ramlan
Othman Iekhsan
author_sort Noor Suriani M
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Highly pathogenic Avian Influenza (HPAI) virus is able to infect many hosts and the virus replicates in high levels in the respiratory tract inducing severe lung lesions. The pathogenesis of the disease is actually the outcome of the infection as determined by complex host-virus interactions involving the functional kinetics of large numbers of participating genes. Understanding the genes and proteins involved in host cellular responses are therefore, critical for the elucidation of the mechanisms of infection.</p> <p>Methods</p> <p>Differentially expressed transcripts regulated in a H5N1 infections of whole lung organ of chicken, <it>in-vitro </it>chick embryo lung primary cell culture (CeLu) and a continuous Madin Darby Canine Kidney cell line was undertaken. An improved mRNA differential display technique (Gene Fishing™) using annealing control primers that generates reproducible, authentic and long PCR products that are detectable on agarose gels was used for the identification of differentially expressed genes (DEGs). Seven of the genes have been selected for validation using a TaqMan<sup>® </sup>based real time quantitative PCR assay.</p> <p>Results</p> <p>Thirty seven known and unique differentially expressed genes from lungs of chickens, CeLu and MDCK cells were isolated. Among the genes isolated and identified include heat shock proteins, Cyclin D2, Prenyl (decaprenyl) diphosphate synthase, IL-8 and many other unknown genes. The quantitative real time RT-PCR assay data showed that the transcription kinetics of the selected genes were clearly altered during infection by the Highly Pathogenic Avian Influenza virus.</p> <p>Conclusion</p> <p>The Gene Fishing™ technique has allowed for the first time, the isolation and identification of sequences of host cellular genes regulated during H5N1 virus infection. In this limited study, the differentially expressed genes in the three host systems were not identical, thus suggesting that their responses to the H5N1 infection may not share similar mechanisms and pathways.</p>
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spelling doaj.art-80486ebaa77342ba87db18c0f7e2e4ea2022-12-21T19:10:39ZengBMCVirology Journal1743-422X2011-04-018119610.1186/1743-422X-8-196Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systemsNoor Suriani MMohamed MaizanOmar Abdul RHassan Sharifah SBalasubramaniam Vinod RMTMohamed RamlanOthman Iekhsan<p>Abstract</p> <p>Background</p> <p>Highly pathogenic Avian Influenza (HPAI) virus is able to infect many hosts and the virus replicates in high levels in the respiratory tract inducing severe lung lesions. The pathogenesis of the disease is actually the outcome of the infection as determined by complex host-virus interactions involving the functional kinetics of large numbers of participating genes. Understanding the genes and proteins involved in host cellular responses are therefore, critical for the elucidation of the mechanisms of infection.</p> <p>Methods</p> <p>Differentially expressed transcripts regulated in a H5N1 infections of whole lung organ of chicken, <it>in-vitro </it>chick embryo lung primary cell culture (CeLu) and a continuous Madin Darby Canine Kidney cell line was undertaken. An improved mRNA differential display technique (Gene Fishing™) using annealing control primers that generates reproducible, authentic and long PCR products that are detectable on agarose gels was used for the identification of differentially expressed genes (DEGs). Seven of the genes have been selected for validation using a TaqMan<sup>® </sup>based real time quantitative PCR assay.</p> <p>Results</p> <p>Thirty seven known and unique differentially expressed genes from lungs of chickens, CeLu and MDCK cells were isolated. Among the genes isolated and identified include heat shock proteins, Cyclin D2, Prenyl (decaprenyl) diphosphate synthase, IL-8 and many other unknown genes. The quantitative real time RT-PCR assay data showed that the transcription kinetics of the selected genes were clearly altered during infection by the Highly Pathogenic Avian Influenza virus.</p> <p>Conclusion</p> <p>The Gene Fishing™ technique has allowed for the first time, the isolation and identification of sequences of host cellular genes regulated during H5N1 virus infection. In this limited study, the differentially expressed genes in the three host systems were not identical, thus suggesting that their responses to the H5N1 infection may not share similar mechanisms and pathways.</p>http://www.virologyj.com/content/8/1/196Avian Influenza virusmRNA differential displayannealing control primerhsp60 genecyclin D2 geneInterleukin 8 gene
spellingShingle Noor Suriani M
Mohamed Maizan
Omar Abdul R
Hassan Sharifah S
Balasubramaniam Vinod RMT
Mohamed Ramlan
Othman Iekhsan
Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
Virology Journal
Avian Influenza virus
mRNA differential display
annealing control primer
hsp60 gene
cyclin D2 gene
Interleukin 8 gene
title Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
title_full Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
title_fullStr Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
title_full_unstemmed Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
title_short Cellular transcripts regulated during infections with Highly Pathogenic H5N1 Avian Influenza virus in 3 host systems
title_sort cellular transcripts regulated during infections with highly pathogenic h5n1 avian influenza virus in 3 host systems
topic Avian Influenza virus
mRNA differential display
annealing control primer
hsp60 gene
cyclin D2 gene
Interleukin 8 gene
url http://www.virologyj.com/content/8/1/196
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