Alpha‐1‐antitrypsin and complement component C7 are involved in asthma exacerbation

Abstract

Asthma is the most common chronic disorder in childhood and asthma exacerbation is an important cause of childhood morbidity and hospitalization. Allergic responses are known to be biased toward T-helper type 2 in asthmatics; however, the pathogenesis of asthma is not simple, and our understanding of the disease mechanism remains incomplete. The aim of the present study was to identify protein expression signatures that reflect acute exacerbation of asthma. Plasma was taken twice from pediatric asthmatic patients, once during asthma exacerbation and once during a stable period. Plasma was also taken from healthy children as a control. The protein profiles of plasma during asthma exacerbation were analyzed by 2-DE and 49 spots were differentially expressed during asthma exacerbation. Thirty-eight of the spots were successfully identified by MALDI-TOF MS. Proteins up- or down-regulated during asthma exacerbation were involved in responses to stress and pathogens, in the complement and coagulation cascades, and in acute-phase responses. Among the differentially expressed proteins, up-regulation of alpha-1-antitrypsin and complement component C7 was confirmed by nephelometry and ELISA. Our present results suggest that protease inhibitors and complement components may be involved in asthma exacerbation, and plasma level of alpha-1-antitrypsin may be a potential biomarker for asthma.

Apolipoprotein A-IV is a candidate target molecule for the treatment of seasonal allergic rhinitis

“…These excised gels were destained with destaining solution containing 15 mmol/L potassium hexacyanoferrate (III) (Wako, Osaka, Japan) and 50 mmol/L sodium thiosulfate (Sigma, St Louis, Mo) and digested with sequencing-grade modified trypsin (Promega, Madison, Wis), and using these peptide extracts, we performed nano-HPLC sample separation with a Paradigm MS4 LC system (Michrom BioResources, Auburn, Calif) 14 and MALDI-TOF/TOF mass spectrometry with ultraflex II (BRUKER Daltonics, Billerica, Mass). 13 Molecular mass information obtained by MALDI-TOF/ TOF mass spectrometry was searched against the Swiss-Prot protein database (version 56.5, http://www.expasy.org/sprot/) with the MASCOT search program (version 2.2; MatrixScience, Boston, Mass) automatically using the Warp-LC software (BURUKER Daltnics), which attached to the ultraflex II. The following search criteria were used: (1) the taxonomy was Homo sapiens (human being); (2) the specified enzyme was trypsin, with up to 1 missed cleavage permitted; (3) the fixed modifications were carbamidomethylation of cysteine residues and variable modifications were oxidation of methionine residues; and (4) the peptide tolerance and MS/MS tolerance were set at 100 ppm and 6 0.5 d, respectively.…”

“…13 Labeled proteins were visualized with a Typhoon 9400 Imager (GE Healthcare), and 24 images of paired samples were analyzed with the DeCyder Software Platform version 4.0 (GE Healthcare). The detailed Methods are available in this article's Online Repository at www.jacionline.org.…”

Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) – an Introduction for Biologists

“…A recent proteomic study has also shown that levels of haptoglobin are elevated in BAL fluid in patients with mild asthma, and reported that haptoglobin may play a role in the differentiation of fibroblast progenitor cells, suggesting a novel role for haptoglobin in airway remodeling in patients with asthma [20]. Conversely Nishioka et al showed that the serum concentration of haptoglobin was decreased during acute exacerbation in asthmatic children [21]. Haptoglobin isoforms and fragments are also elevated in plasma of severe acute respiratory syndrome (SARS) patients [22].…”

Plasma proteomic profile of sulfur mustard exposed lung diseases patients using 2-dimensional gel electrophoresis

“…Molecular weight (MW) and pI are theoretical and * indicates proteins present on the gel at a MW or pI different to these values novel role for haptoglobin in airway remodeling in patients with asthma [20]. Conversely Nishioka et al showed that the serum concentration of haptoglobin was decreased during acute exacerbation in asthmatic children [21]. Haptoglobin isoforms and fragments are also elevated in plasma of severe acute respiratory syndrome (SARS) patients [22].…”