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

Author:

Zhang Fuquan1,Hopwood Paul2,Abrams Charles C.1,Downing Alison3,Murray Frazer3,Talbot Richard3,Archibald Alan3,Lowden Stewart2,Dixon Linda K.1

Affiliation:

1. Institute for Animal Health Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 ONF, United Kingdom

2. ARK-Genomics Roslin Institute, Roslin, Midlothian EH25 9PS, United Kingdom

3. Royal Dick Veterinary College, University of Edinburgh, Summerhall, Edinburgh EH9 1QH, United Kingdom

Abstract

ABSTRACT 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 1β 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.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

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