Proteome changes of lungs artificially infected with H-PRRSV and N-PRRSV by two-dimensional fluorescence difference gel electrophoresis

Abstract

BackgroundPorcine reproductive and respiratory syndrome with PRRS virus (PRRSV) infection, which causes significant economic losses annually, is one of the most economically important diseases affecting swine industry worldwide. In 2006 and 2007, a large-scale outbreak of highly pathogenic porcine reproductive and respiratory syndrome (PRRS) happened in China and Vietnam. However little data is available on global host response to PRRSV infection at the protein level, and similar approaches looking at mRNA is problematic since mRNA levels do not necessarily predict protein levels. In order to improve the knowledge of host response and viral pathogenesis of highly virulent Chinese-type PRRSV (H-PRRSV) and Non-high-pathogenic North American-type PRRSV strains (N-PRRSV), we analyzed the protein expression changes of H-PRRSV and N-PRRSV infected lungs compared with those of uninfected negative control, and identified a series of proteins related to host response and viral pathogenesis.ResultsAccording to differential proteomes of porcine lungs infected with H-PRRSV, N-PRRSV and uninfected negative control at different time points using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mass spectrometry identification, 45 differentially expressed proteins (DEPs) were identified. These proteins were mostly related to cytoskeleton, stress response and oxidation reduction or metabolism. In the protein interaction network constructed based on DEPs from lungs infected with H-PRRSV, HSPA8, ARHGAP29 and NDUFS1 belonged to the most central proteins, whereas DDAH2, HSPB1 and FLNA corresponded to the most central proteins in those of N-PRRSV infected.ConclusionsOur study is the first attempt to provide the complex picture of pulmonary protein expression during H-PRRSV and N-PRRSV infection under the in vivo environment using 2D-DIGE technology and bioinformatics tools, provides large scale valuable information for better understanding host proteins-virus interactions of these two PRRSV strains.

Identification of Differentially Expressed Proteins in Porcine Alveolar Macrophages Infected with Virulent/Attenuated Strains of Porcine Reproductive and Respiratory Syndrome Virus

“…Our study showed that HSPB1 in PAM cells was up- and down-regulated by the virulent HuN4 strain and the attenuated HuN4-F112 strain, respectively, which confirmed related previous studies [18] , [20] . In addition, infections involving many other viruses, such as the avian influenza H9N2 [38] , the Epstein-Barr virus [39] , the African swine fever virus [40] , IBDV [41] and CSFV [42] also induce upregulation of HSP27 expression.…”

“…Ding et al [19] utilized the JL/07/SW strain (HP-PRRSV) to infect PAM cells in vitro and detected 29 differentially expressed proteins. Xiao et al [20] identified 45 differentially expressed proteins using the virulent Chinese strain and the attenuated North American strain to infect pigs in vivo and then compared the proteomic changes induced by the virulent/attenuated strain in lung tissue. However, the specific proteins involved in viral replication in these infections remain unknown.…”

An interactome map of the nucleocapsid protein from a highly pathogenic North American porcine reproductive and respiratory syndrome virus strain generated using SILAC‐based quantitative proteomics

“…This dataset can also be compared to other proteomic approaches that have been applied to the study of PRRSV interactions with the host cell and virus particles, which include changes in the abundance of proteins involved in translation . To our knowledge, this is the first application of SILAC coupled to GFP‐Trap pull‐downs for the analysis of a viral protein and mapping an interactome.…”

Pig proteomics: A review of a species in the crossroad between biomedical and food sciences

“…A recent study from Xiao et al compared the proteome patterns of lung samples taken from pigs infected with either a highly virulent Chinese strain of PRRSV, a less virulent North American strain, or with no infection, using 2D-DIGE. Also these authors found mainly changes in the central metabolic pathways, and found very few strain specific changes, but including however the observation that different types of heat shock proteins were up regulated as a response to these two different strains of PRRSV [103]. In a more recent study these authors also presented a genome wide transcriptome analysis of the host response to the North American strain, using deep-sequencing technology which provides the so far most sensitive detection of gene regulation possible [104].…”