Identification of tuberculosis-associated proteins in whole blood supernatant

Takahiro Tanaka, Shinsaku Sakurada, Keiko Kano, Eri Takahashi, Kazuki Yasuda, Hisashi Hirano, Yasushi Kaburagi, Nobuyuki Kobayashi, Nguyen T. Le Hang, Luu T. Lien, Ikumi Matsushita, Minako Hijikata, Takafumi Uchida, Naoto Keicho

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

Background: Biological parameters are useful tools for understanding and monitoring complicated disease processes. In this study, we attempted to identify proteins associated with active pulmonary tuberculosis (TB) using a proteomic approach.Methods: To assess TB-associated changes in the composition of human proteins, whole blood supernatants were collected from patients with active TB and healthy control subjects. Two-dimensional difference gel electrophoresis (2D-DIGE) was performed to analyze proteins with high molecular weights (approximately >20 kDa). Baseline protein levels were initially compared between patients with active TB and control subjects. Possible changes of protein patterns in active TB were also compared ex vivo between whole blood samples incubated with Mycobacterium tuberculosis (Mtb)-specific antigens (stimulated condition) and under unstimulated conditions. Immunoblot and enzyme-linked immunosorbent assays (ELISA) were performed to confirm differences in identified proteins.Results: Under the baseline condition, we found that the levels of retinol-binding protein 4 (RBP4), fetuin-A (also called α-HS-glycoprotein), and vitamin D-binding protein differed between patients with active TB and control subjects on 2D gels. Immunoblotting results confirmed differential expression of RBP4 and fetuin-A. ELISA results further confirmed significantly lower levels of these two proteins in samples from patients with active TB than in control subjects (P < 0.0001). Mtb-specific antigen stimulation ex vivo altered clusterin expression in whole blood samples collected from patients with active TB.Conclusions: We identified TB-associated proteins in whole blood supernatants. The dynamics of protein expression during disease progression may improve our understanding of the pathogenesis of TB.

Original languageEnglish
Article number71
JournalBMC infectious diseases
Volume11
DOIs
Publication statusPublished - 2011 Mar 22

ASJC Scopus subject areas

  • Infectious Diseases

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