DPP4, the Middle East Respiratory Syndrome Coronavirus Receptor, is Upregulated in Lungs of Smokers and Chronic Obstructive Pulmonary Disease Patients

Leen J M Seys, W Widagdo, Fien M Verhamme, Alex Kleinjan, Wim Janssens, Guy F Joos, Ken R Bracke, Bart L Haagmans, Guy G Brusselle, Leen J M Seys, W Widagdo, Fien M Verhamme, Alex Kleinjan, Wim Janssens, Guy F Joos, Ken R Bracke, Bart L Haagmans, Guy G Brusselle

Abstract

Background: Middle East respiratory syndrome coronavirus (MERS-CoV) causes pneumonia with a relatively high case fatality rate in humans. Smokers and chronic obstructive pulmonary disease (COPD) patients have been reported to be more susceptible to MERS-CoV infection. Here, we determined the expression of MERS-CoV receptor, dipeptidyl peptidase IV (DPP4), in lung tissues of smokers without airflow limitation and COPD patients in comparison to nonsmoking individuals (never-smokers).

Methods: DPP4 expression was measured in lung tissue of lung resection specimens of never-smokers, smokers without airflow limitation, COPD GOLD stage II patients and in lung explants of end-stage COPD patients. Both control subjects and COPD patients were well phenotyped and age-matched. The mRNA expression was determined using qRT-PCR and protein expression was quantified using immunohistochemistry.

Results: In smokers and subjects with COPD, both DPP4 mRNA and protein expression were significantly higher compared to never-smokers. Additionally, we found that both DPP4 mRNA and protein expression were inversely correlated with lung function and diffusing capacity parameters.

Conclusions: We provide evidence that DPP4 is upregulated in the lungs of smokers and COPD patients, which could partially explain why these individuals are more susceptible to MERS-CoV infection. These data also highlight a possible role of DPP4 in COPD pathogenesis.

Keywords: Middle East respiratory syndrome coronavirus (MERS-CoV); chronic obstructive pulmonary disease (COPD); dipeptidyl peptidase 4 (DPP4); smoking.

© The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

Figures

Figure 1.
Figure 1.
DPP4 mRNA expression in the lung tissues of smokers and COPD patients. A, DPP4 mRNA expression was measured by qRT-PCR and normalized to three reference genes (GAPDH, HPRT-1, SDHA). DPP4 mRNA expression in the lungs of smokers and COPD patients is significantly higher in comparison to that of never smokers. B, Correlation of DPP4 mRNA expression with post-bronchodilator FEV1 values. C, Correlation of DPP4 mRNA expression with post-bronchodilator Tiffeneau index (FEV1/FVC). D, Correlation of DPP4 mRNA expression with DLCO (diffusing capacity or transfer factor of the lung for carbon monoxide). E, Correlation of DPP4 mRNA expression with KCO (carbon monoxide transfer coefficient). **P < .01, ***P < .001. Abbreviations: COPD, chronic obstructive pulmonary disease; DLCO, diffusing capacity of the lung for carbon monoxide; FEV/FVC, forced expiratory volume in 1 second/forced vital capacity; GOLD, global initiative for obstructive lung disease; KCO, transfer of carbon monoxide coefficient; mRNA, messenger RNA; qRT-PCR, quantitative reverse-transcription polymerase chain reaction.
Figure 2.
Figure 2.
DPP4 protein expression in the bronchiolar epithelium and the alveolar tissues of never smoker, smoker, and COPD patients. Representative images of DPP4 staining in the bronchiolar epithelium (top row) and alveoli (middle and bottom row) of A,E,I, never-smoker, B,F,J, smoker without airflow limitation, C,G,K, subject with COPD GOLD stage II and D,H,L, subject with COPD GOLD stage III–IV. I–-L, are immunohistochemical stainings of DPP4 (brown) and aquaporin 5 (marker of type I alveolar epithelial cells) and pro-surfactant C (marker of type II alveolar epithelial cells) (both in blue). Co-staining of DPP4 with either one of the alveolar epithelial cell types results in a dark brown stain. DPP4 was mainly expressed in the alveolar epithelial cells and expressed the most intense in the COPD GOLD stage III–IV group. A 400× magnification was used for all photomicrographs in this figure. Abbreviation: COPD, chronic obstructive pulmonary disease; GOLD, global initiative for chronic obstructive lung disease.
Figure 3.
Figure 3.
DPP4 staining in the proximal bronchi epithelium. Representative images of DPP4 staining in proximal bronchial epithelium and submucosal glands of the healthy control subject with COPD GOLD stage II. DPP4 was hardly detected in the apical surface of the proximal bronchi epithelium of both healthy control and COPD patients. Submucosal glands here served as positive control for DPP4 staining. Abbreviation: COPD, chronic obstructive pulmonary disease.
Figure 4.
Figure 4.
DPP4 protein expression in the lung tissues of smokers and COPD patients. , DPP4 protein expression was analyzed by using Axiovision software (Zeiss). The area of DPP4 positive signal was normalized to the total area of cells present in each analyzed image. DPP4 protein expression in the lungs of smokers and COPD patients is significantly higher in comparison to that of never smokers. B, Correlation of alveolar DPP4 protein expression with post-bronchodilator FEV1 values. C, Correlation of alveolar DPP4 protein expression with post-bronchodilator Tiffeneau index (FEV1/FVC). D, Correlation of alveolar DPP4 protein expression with DLCO (diffusing capacity or transfer factor of the lung for carbon monoxide). E, Correlation of alveolar DPP4 protein expression with KCO (carbon monoxide transfer coefficient). **P < .01, ***P < .001. Abbreviation: COPD, chronic obstructive pulmonary disease.

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