J Vet Sci.  2014 Dec;15(4):511-517. 10.4142/jvs.2014.15.4.511.

Proteomic analysis of chicken peripheral blood mononuclear cells after infection by Newcastle disease virus

• Deng X
• Cong Y
• Yin R
• Yang G
• Ding C
• Yu S
• Liu X
• Wang C
• Ding Z

• KMID: 2070237
• DOI: http://doi.org/10.4142/jvs.2014.15.4.511

Abstract

Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.

Keyword

Newcastle disease virus; peripheral blood mononuclear cells; proteomics

MeSH Terms

Animals
Avian Proteins/*genetics/metabolism
*Chickens
*Gene Expression Regulation
Leukocytes, Mononuclear/enzymology/virology
Newcastle Disease/*genetics/virology
Newcastle disease virus/*physiology
Poultry Diseases/*genetics/virology
*Proteome
Specific Pathogen-Free Organisms
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/veterinary
Tandem Mass Spectrometry/veterinary
Avian Proteins
Proteome

Figure

• 

  Fig. 1 RT-PCR results for specific detection of partial Newcastle disease virus (NDV) M gene. Lane 1, infected group; Lane 2, mock-infected group; Lane M, DL2000 marker.

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  Fig. 2 Representative 2-dimensional electrophoresis (DE) gel images obtained for peripheral blood mononuclear cells (PBMCs) from chickens that were infected by NDV or mock-infected. (A) Mock-infected control group and (B) day 5 post-infected group. The protein spots were visualized by silver staining. MM denotes the molecular mass.

• 

  Fig. 3 Validation of altered lamin A and integrin-linked kinase (ILK) expression by Western blot analysis. (A) Magnified 2-DE images of lamin A and ILK for the mock-infected control and NDV-infected groups. (B) Western blot analysis of lamin A and ILK expression.

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