Macrophage transcriptional responses following in vitro infection with a highly virulent African swine fever virus isolate.
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 1beta 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.
Zhang
Fuquan
F
Institute for Animal Health Pirbright Laboratory, Ash Road, Pirbright, Woking, Surrey GU24 ONF, United Kingdom.
Hopwood
Paul
P
Abrams
Charles C
CC
Downing
Alison
A
Murray
Frazer
F
Talbot
Richard
R
Archibald
Alan
A
Lowden
Stewart
S
Dixon
Linda K
LK
D000358:African Swine Fever Virus / Q000302:isolation & purification / Q000472:pathogenicity*; D000818:Animals; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D053583:Interleukin-1beta / Q000096:biosynthesis / Q000235:genetics; D016676:Macrophages, Alveolar / Q000276:immunology / Q000378:metabolism* / Q000821:virology*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D034421:Sus scrofa; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics; D014774:Virulence
D053583:Interleukin-1beta; D014409:Tumor Necrosis Factor-alpha