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Journal of Virology, November 2006, p. 10514-10521, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.00485-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Macrophage Transcriptional Responses following In Vitro Infection with a Highly Virulent African Swine Fever Virus Isolate

Fuquan Zhang,1,# Paul Hopwood,2,# Charles C. Abrams,1 Alison Downing,3 Frazer Murray,3 Richard Talbot,3 Alan Archibald,3 Stewart Lowden,2 and Linda K. Dixon1*

Institute for Animal Health Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 ONF, United Kingdom,1 ARK-Genomics Roslin Institute, Roslin, Midlothian EH25 9PS, United Kingdom,2 Royal Dick Veterinary College, University of Edinburgh, Summerhall, Edinburgh EH9 1QH, United Kingdom3

Received 8 March 2006/ Accepted 31 July 2006

We used a porcine microarray containing 2,880 cDNAs to investigate the response of macrophages to infection by a virulent African swine fever virus (ASFV) isolate, Malawi LIL20/1. One hundred twenty-five targets were found to be significantly altered at either or both 4 h and 16 h postinfection compared with targets after mock infection. These targets were assigned into three groups according to their temporal expression profiles. Eighty-six targets showed increased expression levels at 4 h postinfection but returned to expression levels similar to those in mock-infected cells at 16 h postinfection. These encoded several proinflammatory cytokines and chemokines, surface proteins, and proteins involved in cell signaling and trafficking pathways. Thirty-four targets showed increased expression levels at 16 h postinfection compared to levels at 4 h postinfection and in mock-infected cells. One host gene showed increased expression levels at both 4 and 16 h postinfection compared to levels in mock-infected cells. The microarray results were validated for 12 selected genes by quantitative real-time PCR. Levels of protein expression and secretion were measured for two proinflammatory cytokines, interleukin 1ß and tumor necrosis factor alpha, during a time course of infection with either the virulent Malawi LIL20/1 isolate or the OUR T88/3 nonpathogenic isolate. The results revealed differences between these two ASFV isolates in the amounts of these cytokines secreted from infected cells.

* Corresponding author. Mailing address: Institute for Animal Health Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 ONF, United Kingdom. Phone: 44 1483 231062. Fax: 44 1483 232448. E-mail: linda.dixon{at}bbsrc.ac.uk.

# F. Zhang and P. Hopwood contributed equally to this work.

Journal of Virology, November 2006, p. 10514-10521, Vol. 80, No. 21
0022-538X/06/$08.00+0     doi:10.1128/JVI.00485-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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