Proteomic analysis of human brain microvascular endothelial cells reveals differential protein expression in response to enterovirus 71 infection.

Wenying Luo,Wei Zhao,Hong Cao,Wenxu Hong,Liang Peng,Jiayu Zhong,Sheng He Huang,Jianjun Liu,Renli Zhang

doi:10.1155/2015/864169

Abstract

2D DIGE technology was employed on proteins prepared from human brain microvascular endothelial cells (HBMEC), to study the differentially expressed proteins in cells at 0 h, 1 h, 16 h, and 24 h after infection. Proteins found to be differentially expressed were identified with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDITOF/TOF MS) analysis. We identified 43 spots showing changes of at least 2.5 fold up- or downregulated expressions in EV71-infected cells at different time when comparing to control, and 28 proteins could be successfully identified by MALDI TOF/TOF mass spectrometry analysis. 4 proteins were significantly upregulated, and 6 proteins were downregulated, another 18 proteins were different expression at different incubation time. We identified changes in the expression of 12 cellular metabolism-related proteins, 5 molecules involved in cytoskeleton, 3 molecules involved in energy metabolism, 2 molecules involved in signal transduction, 1 molecule involved in the ubiquitin-proteasome pathway, 1 molecule involved in cell cycle, 1 molecule involved in apoptosis-related protein, 1 molecular chaperone, and 2 unknown proteins. These findings build up a comprehensive profile of the HBMEC proteome and provide a useful basis for further analysis of the pathogenic mechanism that underlies EV71 infections to induce severe neural complications.

Highlights

Human enterovirus 71 (EV71) was first described during an outbreak with central nervous system complications in 1974 [1], which is a small, nonenveloped positive- stranded RNA virus that belonging to the enterovirus genus of the Picornaviridae family [2]
EV71 is a major pathogen of hand-foot-andmouth disease (HFMD); there were greater numbers of fatal cases of HFMD with symptoms of central nervous system (CNS) occurred in Taiwan [3], western Australia [4], Malaysia [5], Japan [6], Singapore [7], and South Korea [8] during the last decade
The four upregulated proteins were identified as protein expression at 1 hpi, 16 hpi, and 24 hpi upregulated after EV71 infection, which included splicing factor arginine/serinerich 2, ubiquinol-cytochrome c reductase core I protein, glutathione synthetase, and actin cytoplasmic 1

Summary

Introduction

Human enterovirus 71 (EV71) was first described during an outbreak with central nervous system complications in 1974 [1], which is a small, nonenveloped positive- stranded RNA virus that belonging to the enterovirus genus of the Picornaviridae family [2]. Many data show that EV71 infection has a complex pathogenesis and that the central nervous system (CNS) is likely the main target of the EV71 virus [10]. The human brain microvascular endothelial cells (HBMEC) are a special type of cell that constitute the BBB [17]. We found EV71-infected HBMEC can induce 28 differently expressed protein spots compared with control. These proteins might affect BBB dysfunction and lead to the development of EV71 CNS disease

Material and Methods

Comparative Proteome Analyses of HBMEC

Results and Discussion