title: Identification of tuberculosis-associated proteins in whole blood supernatant.
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.
authors.affiliation: Molecular Enzymology, Graduate School of Agricultural Science, Tohoku University, 1-1 Amamiya-machi, Tsutsumidori, Aoba-ku, Sendai, Miyagi 981-8555, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tanaka
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Sakurada
authors.forename: Keiko
authors.initials: K
authors.lastname: Kano
authors.forename: Eri
authors.initials: E
authors.lastname: Takahashi
authors.forename: Kazuki
authors.initials: K
authors.lastname: Yasuda
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hirano
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kaburagi
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Nguyen Thi Le
authors.initials: NT
authors.lastname: Hang
authors.forename: Luu Thi
authors.initials: LT
authors.lastname: Lien
authors.forename: Ikumi
authors.initials: I
authors.lastname: Matsushita
authors.forename: Minako
authors.initials: M
authors.lastname: Hijikata
authors.forename: Takafumi
authors.initials: T
authors.lastname: Uchida
authors.forename: Naoto
authors.initials: N
authors.lastname: Keicho
mesh_terms: D000328:Adult; D000368:Aged; D000942:Antigens, Bacterial; D001798:Blood Proteins / Q000032:analysis*; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D015151:Immunoblotting; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D040901:Proteomics / Q000379:methods*; D054839:Retinol-Binding Proteins, Plasma / Q000032:analysis; D014397:Tuberculosis, Pulmonary / Q000097:blood*; D057825:Two-Dimensional Difference Gel Electrophoresis; D014809:Vitamin D-Binding Protein / Q000097:blood; D055815:Young Adult; D060749:alpha-2-HS-Glycoprotein
chemical_list: C556354:AHSG protein, human; D000942:Antigens, Bacterial; D001798:Blood Proteins; C101326:RBP4 protein, human; D054839:Retinol-Binding Proteins, Plasma; D014809:Vitamin D-Binding Protein; D060749:alpha-2-HS-Glycoprotein; C072681:Mycobacterium tuberculosis antigens