J Korean Med Sci.  2020 Nov;35(43):e362. 10.3346/jkms.2020.35.e362.

Serum Calprotectin Is a Potential Marker in Patients with Asthma

• Lee YG
• Hong Js
• Lee PH
• Lee Jh
• Park SW
• Kim DJ
• Jang AS

• KMID: 2508500
• DOI: http://doi.org/10.3346/jkms.2020.35.e362

Abstract

Background
Calprotectin is the major cytosolic protein in neutrophil granulocytes. Although asthma is known to cause eosinophilic inflammation, some patients with asthma have non-eosinophilic inflammation, which is characterized by local neutrophilic inflammation. The aim of this study was to assess calprotectin expression levels in a mouse model of asthma, and to observe the relationship of serum calprotectin level and clinical variables in patients with asthma.
Methods
Mice were sensitized and challenged with 10 μg and 20 μg of Aspergillus fumigatus, respectively; mice treated with saline were used as a control. The levels of calprotectin were determined using enzyme-linked immunosorbent assay, immunoblotting, and immunohistochemical analysis. The serum levels of calprotectin were also assessed in patients with asthma. The relationship between calprotectin and clinicopathological characteristics was determined.
Results
Calprotectin, S100A8, and S100A9 expression was elevated in the mouse lungs, Calprotectin levels were higher in the serum of patients with asthma (n = 33) compared with those of healthy individuals (n = 28). Calprotectin levels correlated with forced expiratory volume in one second/forced vital capacity (r = −0.215, P = 0.043), smoke amount (r = 0.413, P = 0.017), body mass index (r = −0.445,P= 0.000), and blood neutrophil percentage (r = 0.300, P = 0.004) in patients with asthma.
Conclusion
Our data suggest that calprotectin could potentially be used as a biomarker for asthma.

Keyword

Calprotectin; Asthma; Airway Inflammation

Figure

• 

  Fig. 1 AHR and inflammation in Aspergillus fumigatus sensitization/challenge-mice. (A) A. fumigatus sensitization and challenge protocol. (B) AHR was more increased in A. fumigatus-mice compared to control mice. (C) Total and differential cell count in BALF. BALF was collected on 25 day. Total and differential cell count was increased in A. fumigatus sensitization and challenge mice compared to control mice.AHR = airway hyperresponsiveness, BALF = bronchoalveolar lavage fluid, I.P. = intraperitoneal, I.N. = intranasal, BAL = bronchoalveolar lavage.*P < 0.05, A. fumigatus vs. Sham.

• 

  Fig. 2 H&E staining, PAS and IHC staining of mouse lung paraffin sections. (A) Top panel: H&E-stained lung tissue contained more inflammatory cell infiltrations in Aspergillus fumigatus-sensitization/challenge mice than control mice. Middle panel: PAS-stained airways contained increased goblet cells in the A. fumigatus-sensitization/challenge mice. Lower panel: IHC-stained lung tissue contained increased calprotectin in A. fumigatus sensitization/challenge mice. (B) The quantitation of goblet cells counts in mouse lung. (C) The quantitation of calprotectin in mouse lung.H&E = hematoxylin and eosin, PAS = periodic acid-Schiff, IHC = immunohistochemical.*P < 0.05, Fungus vs. Sham.

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  Fig. 3 Lung protein level of calprotectin as determined by Western blot. Densitometry was determined with 3 immunoblots and normalized to β-actin. Bar graphs are densitometric data (means ± standard deviation).*P < 0.05 compared to controls.

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  Fig. 4 Calprotectin level in control subjects, stable, and exacerbated asthmatic patients.*P < 0.05, stable and exacerbated asthma vs. control subjects.

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  Fig. 5 Relationship of calprotectin with WBC, smoke amount, neutrophil proportion, BMI, FEV1%pred. FEV1/FVC, and Log PC20.WBC = white blood cell, BMI = body mass index, FEV1 = forced expiratory volume in one second, FVC = forced vital capacity, %pred. = % predicted, PC20 = the concentration of methacholine required to decrease the forced expiratory volume in one second by 20%.

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