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Original Research: ASTHMA |

Protein Microarray Analysis in Patients With Asthma: Elevation of the Chemokine PARC/CCL18 in Sputum

Hyo-Bin Kim, MD, PhD; Chang-Keun Kim, MD, PhD; Koji Iijima, PhD; Takao Kobayashi, PhD; Hirohito Kita, MD
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*From the Department of Pediatrics (Drs. H-B Kim and C-K Kim), Asthma & Allergy Center, Inje University Sanggye Paik Hospital, Seoul, Korea; and the Department of Immunology (Drs. Iijima, Kobayashi, and Kita), Allergic Diseases Research Laboratory, Mayo Clinic and Foundation, Rochester, MN.

Correspondence to: Hirohito Kita, MD, Department of Immunology, Allergic Diseases Research Laboratory, Mayo Clinic, Guggenheim 301, 200 First St SW, Rochester, MN 55905; e-mail: kita.hirohito@mayo.edu

†These authors contributed equally to this work.


This work was supported by a grant from the National Institutes of Health (AI50494) and in part by the Korean Research Foundation grant funded by the Korean Government (MOEHRD) [KRF-2005-042-E00078].

The authors have reported to the ACCP that no significant conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article.

Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (www.chestjournal.org/misc/reprints.shtml).


Chest. 2009;135(2):295-302. doi:10.1378/chest.08-0962
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Background:  Microarray technology offers a new opportunity to gain insight into global gene and protein expression profiles in asthma. To identify novel factors produced in the asthmatic airway, we analyzed sputum samples by using a membrane-based human cytokine microarray technology in patients with bronchial asthma (BA).

Methods:  Induced sputum was obtained from 28 BA subjects, 20 nonasthmatic atopic control (AC) subjects, and 38 nonasthmatic nonatopic normal control (NC) subjects. The microarray samples of subjects were randomly selected from nine BA subjects, three AC subjects, and six NC subjects. Sputum supernatants were analyzed using a custom human cytokine array (RayBio Custom Human Cytokine Array; RayBiotech; Norcross, GA) designed to analyze 79 specific cytokines simultaneously. The levels of growth-regulated oncogene (GRO)-α, eotaxin-2, and pulmonary and activation-regulated chemokine (PARC)/CCL18 were measured by sandwich enzyme-linked immunosorbent assays (ELISAs), and eosinophil-derived neurotoxin (EDN) was measured by radioimmunoassay.

Results:  By microarray, the signal intensities for GRO-α, eotaxin-2, and PARC were significantly higher in BA subjects than in AC and NC subjects (p = 0.036, p = 0.042, and p = 0.033, respectively). By ELISA, the sputum PARC protein levels were significantly higher in BA subjects than in AC and NC subjects (p < 0.0001). Furthermore, PARC levels correlated significantly with sputum eosinophil percentages (r = 0.570, p < 0.0001) and the levels of EDN (r = 0.633, p < 0.0001), the regulated upon activation, normal T cell expressed and secreted cytokine (r = 0.440, p < 0.001), interleukin-4 (r = 0.415, p < 0.01), and interferon-γ (r = 0.491, p < 0.001).

Conclusions:  By a nonbiased screening approach, a chemokine, PARC, is elevated in sputum specimens from patients with asthma. PARC may play important roles in development of airway eosinophilic inflammation in asthma.

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