Proteomic alteration of Marc-145 cells and PAMs after infection by porcine reproductive and respiratory syndrome virus

Abstract

Viral infections usually result in alterations in the host cell proteome, which determine the fate of infected cells and the progress of pathogenesis. To uncover cellular protein responses in porcine reproductive and respiratory syndrome virus (PRRSV), infected pulmonary alveolar macrophages (PAMs) and Marc-145 cells were subjected to proteomic analysis involving two-dimensional electrophoresis (2-DE) followed by MALDI-TOF-MS/MS identification. Altered expression of 44 protein spots in infected cells was identified in 2D gels, of which the 29 characterised by MALDI-TOF-MS/MS included 17 up-regulated and 12 down-regulated proteins. Some of these proteins were further confirmed at the mRNA level using real-time RT-PCR. Moreover, Western blot analysis confirmed the up-regulation of HSP27, vimentin and the down-regulation of galectin-1. Our study is the first attempt to analyze the cellular protein profile of PRRSV-infected Marc-145 cells using proteomics to provide valuable information about the effects of PRRSV-induced alterations on Marc-145 cell function. Further study of the affected proteins may facilitate our understanding of the mechanisms of PRRSV infection and pathogenesis.

Identification of serum proteomic biomarkers for early porcine reproductive and respiratory syndrome (PRRS) infection

“…In a similar study performed with MALDI-TOF [28], where serum samples were analyzed in the first weeks (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) Table 3). Furthermore, two peaks identified in this study (23.162 and 14.843 kDa) were similar to peaks identified elsewhere (corresponding to Heat shock 27 kDa protein 1 [29][30][31] and Galectin 1 [26,31], respectively). In accordance with [31], the identified peak corresponding to Heat shock 27 kDa protein 1 was upregulated, while the peak corresponding to Galectin 1 was down-regulated.…”

“…Furthermore, two peaks identified in this study (23.162 and 14.843 kDa) were similar to peaks identified elsewhere (corresponding to Heat shock 27 kDa protein 1 [29][30][31] and Galectin 1 [26,31], respectively). In accordance with [31], the identified peak corresponding to Heat shock 27 kDa protein 1 was upregulated, while the peak corresponding to Galectin 1 was down-regulated. Thus, these proteins seem to be very interesting and suitable candidates for future investigations.…”

Inhibition of HSP70 reduces porcine reproductive and respiratory syndrome virus replication in vitro

“…Recently, HSP70 was also found to be elevated after PRRSV infection based on transcriptome and proteome approaches [27,30,31]. In this study, we observed that PRRSV infection induced HSP70 expression in vitro (Figures 1 and 8C), implying that HSP70 may play a potential role in PRRSV infection.…”

Determination of the interactome of non-structural protein12 from highly pathogenic porcine reproductive and respiratory syndrome virus with host cellular proteins using high throughput proteomics and identification of HSP70 as a cellular factor for virus replication

“…Although several studies have been focused on the relationship between PRRSV viral proteins such as N [17,34], NSP1 [35,36], NSP2 [37] and whole virion [38,39] and the host cellular proteins, cellular interactome of PRRSV NSP12 has not been reported to the best of our knowledge. Here, high throughput proteomics coupled with immunoprecipitated strategy was used to identify and quantify cellular proteins which potentially interacted with PRRSV NSP12 and participated in viral replication,…”

“…HSP70 chaperones have been reported to be recruited by some other viruses such as respiratory syncytial virus [18,51], hepatitis C virus [52], porcine circovirus type 2 [53], flock house virus [54] and herpes simplex virus type 1 [55] to benefit their infection, and incorporation of HSP70 family in PRRSV virions has also been reported [38,39]. In this study, we discovered that HSP70 affected the stability of NSP12-EGFP ( Figure.6 7), and treatment with MG132, a proteasome inhibitor, could recover the protein level.…”