Plasma proteome of severe acute respiratory syndrome analyzed by two-dimensional gel electrophoresis and mass spectrometry

Abstract

We have investigated the plasma proteome by using 2D gel electrophoresis and MS from patients with severe acute respiratory syndrome (SARS). A complete proteomic analysis was performed on four patients with SARS in different time courses, and a total of 38 differential spots were selected for protein identification. Most of the proteins identified are acute phase proteins, and their presence represents the consequence of serial cascades initiated by SARS-coronavirus infection. There are several proteins that have never been identified in plasma before using 2D gel electrophoresis, among which peroxiredoxin II was chosen for further study by analyzing additional 20 plasma samples from patients with probable and suspected SARS and patients with fever, respectively. The results showed that the level of plasma peroxiredoxin II in patients with SARS is significantly high and could be secreted by T cells. Taken together, our findings indicate that active innate immune responses, along with the oxidation-associated injuries, may play a major role in the pathogenesis of SARS.proteomic techniques ͉ acute phase proteins ͉ peroxiredoxin II S evere acute respiratory syndrome (SARS) was first recognized at the end of 2002 in Guangdong, China, and since then the disease has spread to several countries. By late July 2003, Ͼ8,000 SARS cases and Ͼ700 SARS-related deaths were reported from Ͼ25 countries around the world, and no effective treatment has been established so far. Through extensive studies, the causative agent of SARS has been identified as a human SARS-coronavirus (SARS-CoV) (1-3). Although the complete genome sequence of SARS-CoV, the structure of the main protease (3CL protease), and the possible receptor have been determined (4-7), the pathogenesis of SARS is still not fully understood.Plasma proteins are useful targets for diagnostic, prognostic, and͞or therapeutic development. With proteomic tools available recently, profiling of human plasma proteome becomes more feasible in searching for disease-related markers (8). To explore the possible pathogenetic mechanisms involving the progression of SARS, we have analyzed the plasma proteins of 22 different plasma samples from four SARS patients in different time courses by using 2D gel electrophoresis (2DE) in combination with MS. The results showed that most of the plasma proteins found in patients with SARS are acute phase proteins (APPs) generated by inflammatory reactions during SARS-CoV-induced acute lung injuries. Materials and MethodsHuman Subjects. Four SARS patients with a total of 22 plasma samples were selected for complete proteomic analysis. These patients were infected by SARS-CoV through nosocomial transmission during one major SARS outbreak in one municipal hospital in Taipei. For comparison, plasma samples from six healthy individuals were used as controls. The protein concentration of each sample was determined by the standard Bradford method. The use of these samples was approved by the Review Board of the Tri-Service General Hospital, National Def...

Redox Regulation in the Extracellular Environment

“…In addition, Prx II, a known cytoplasmic form, has also been found in the plasma of multiple sclerosis patients with severe acute respiratory syndrome; T cells may be a source of secreted Prx II in plasma. 146 Another study also found Prx II in plasma and demonstrated it to be an interaction partner of Trx1. 147 Mouse Prx IV, which contains a signal peptide similar to that of human Prx IV, is apparently not secreted and appears to be a cytoplasmic signaling protein, 148 whereas rat Prx IV appears to function as a secreted protein.…”

Biomarkers: Mining the Biofluid Proteome

“…A 2D-PAGE approach was recently applied to compare plasma samples obtained from patients with severe acute respiratory syndrome (SARS) and healthy individuals (25). Twenty-two plasma samples from four different SARS patients were separated by 2D-PAGE using a narrow-range IPG strip (pH 4 -7), and the resulting profiles were compared with those obtained from six healthy plasma samples.…”

Recent advances in blood‐related proteomics

“…By the same technique, truncated forms of a1-antitrypsin were discovered in serum samples of patients presenting with severe acute respiratory syndrome [77]. Chen et al [78] studied four patients with severe acute respiratory syndrome. After 2-DE, a total of 38 differential spots were selected for protein identification, and most of them corresponded to acute phase proteins.…”