Blood Eosinophil Measurements Throughout the Day in Patients With Stable Chronic Obstructive Pulmonary Disease
Conditions: Chronic Obstructive Pulmonary Disease
Sponsor: Universitair Ziekenhuis Brussel

Protocol Clinical Trial  (21/11/2018) 
Title of project 
Blood eosinophil measurements in patients with Chronic Obstructive Pulmonary disease in stable 
state. 
Objective 
To determine the within-day variation of blood eosinophils in patients with Chronic Obstructive 
Pulmonary disease in stable state. 
To determine if there is a correlation between the blood eosinophils and certain clinical parameters. 
Investigators 
I. Van Rossem, E. Vanderhelst, S. Hanon, S. Vincken, K. Jochmans, J. Vandevoorde 
Departments involved in the trial 
Department of Family Medicine and Chronic Care, Vrije Universiteit Brussel 
Department of Internal Medicine, Division of Respiratory Disease, UZ Brussel 
Department of Clinical Biology, UZ Brussel 
Introduction 
Chronic Obstructive Pulmonary Disease (COPD) is a disease which leads to serious morbidity and 
mortality and it is currently the 4th leading cause of death in the world. (1) It is characterised by 
airflow limitation caused by a mixture of small airways disease and parenchymal destruction, the 
relative contributions of which vary from person to person. (2) It is a heterogeneous disease, 
resulting in different phenotypes with varying clinical and pathophysiological characteristics. One of 
these pathophysiological features is chronic airway inflammation and, although eosinophilic 
inflammation is thought to be a typical characteristic of asthma, there is mounting evidence that 
eosinophilic inflammation can also be present in COPD. (3)  
Blood eosinophils are an accessible biomarker for the eosinophilic inflammation in COPD (3). 
Literature suggests a potential theragnostic role for this biomarker, seeing that COPD patients with 
higher levels of blood eosinophils could exhibit a greater responsiveness to corticosteroid treatments 
(4-7) and probably also to anti–IL-5 therapy. (8) Evidence also suggests that this biomarker could 
have a prognostic use for establishing the risk of exacerbations although conflicting results are found. 
(9) 
But before being able to make decisions in the clinical management of patients based on blood 
eosinophils, more needs to be known about the different confounding factors and sources of within-
subject variability of this biomarker. (10) As described by Brusselle et al. eosinophil counts can 
fluctuate due to their short half-life in blood and due to a diurnal rhythm. (11) There is evidence that 
this diurnal variation exists in healthy individuals (12) but, to the best of our knowledge, this has 
never been investigated in COPD patients before. 
Study design 
Prospective interventional study 

 
 
Subjects 
Number and recruitment of subjects 
50 COPD patients (GOLD A+B+C+D). All eligible COPD patients will be recruited at the 
respiratory unit of the UZ Brussel (included the patients admitted for nocturnal oximetry) or 
from family practices.   
Inclusion criteria 
Participant is willing and able to give informed consent for participation in the study. Patients  
are 18 years or older, with the diagnosis of COPD according to GOLD (post-bronchodilator 
Tiffeneau index <0.7), in stable state of the disease and with a smoking history of >10 pack-
years. 
Exclusion criteria 
Clinical diagnosis of asthma 
Use of systemic corticosteroids (oral, intravenous or infiltration up to six weeks before 
inclusion).  
Pregnancy. 
A recent exacerbation of COPD (<4 weeks ago). 
Replacement of subjects 
Inclusion of patients will continue until 50 patients have gone through the entire study protocol. 
 
Restrictions and prohibitions for the subjects 
None. 
Procedures and collected variables 
Upon admission a questionnaire is filled in by the investigator or his representative: age, gender, 
smoking habits, weight, height, current medication, pulmonary function test with bronchodilation, 
mMRC (Modified Medical Research Council) Dyspnea Scale, exacerbation history in the previous 
year. 
Fractional Exhaled Nictric Oxide (FENO) measurement and blood sampling at 08h00, 12h00, 16h00 
(total red blood cell count, total white blood cell count with formula in absolute count and 
percentage of total white blood cells, and platelet count).  
Randomisation/blinding 
 
Not applicable. 
 
Prior and concomitant therapy 
 
Medication prohibited before and during the trial are systemic corticosteroids (oral, intravenous or 
infiltration, up to six weeks before inclusion). All other medication use is allowed. 
 

Study analysis 
 
Sample size calculation 
Due to the lack of previous research, a sample size calculation was impossible to perform and an 
arbitrary decision was made. 
 
Analysis of the samples 
The analysis of the blood samples will be done at the Department of Clinical Biology, UZ Brussel. The 
pulmonary function test will be performed at the Department of Internal Medicine, Division of 
Respiratory Disease, UZ Brussel.  
Statistical analysis 
Statistical analyses will be performed by the Department of Statistics and Data-analyses of the Vrije 
Universiteit Brussel. 
 
Quality control and quality assurance 
 
Quality control and quality assurance will be done according to guidelines of the Association for the 
Accreditation of Human Research Protection Programs. 
 
Data Protection 
 
Responsible for processing of personal data 
Department of Family Medicine and Chronic Care, Vrije Universiteit Brussel 
Contact person: Inès Van Rossem, Vakgroep Huisartsgeneeskunde en Chronische Zorg, Vrije 
Universiteit Brussel, Laarbeeklaan 103, 1090 Jette, tel: +32 2 477 43 11 
 
Data Protection Officer (DPO) 
Audrey van Scharen, DPO Vrije Universiteit Brussel (DPO@vub.be) 
 
Goal of the processing 
The gathered data will be used for the academic scientific research purposes described in this protocol. 
It will be processed according to the principles imposed by the European General Data Protection 
Regulation (GDPR), which has been in force since 25 May 2018.  
The legal basis for the processing is permission. There is an explicit permission from the data subject 
for the processing of personal data through informed consent. Permission can be withdrawn by the 
participant at any time, for any reason and without having to state a reason. 
 
Recipients of data 
The prime investigator and the study nurse will be the only individuals with access to personnel data. 
Study data will not be transferred to other countries. 
 
Storage 
The study data will be stored in an excel file in a coded fashion with the encryption key held in a 
separate file. Both the data file and the encryption key will be password protected and saved in the 
one drive system of the Vrije Universiteit Brussel. 
The gathered study data will be saved for a period of minimum 20 years. This in accordance with the 
Belgian law on experiments on the human person (7 May 2004). 
 
 
Publication policy 
 

All authors will contribute to publication and hold all publication rights. 
 
Funding 
A request for funding was submitted to the “Belgische Vereniging voor Pneumologie”. 
 
References 
1. 
Lozano R, Naghavi M, Foreman K, et al. Global and regional mortality from 235 causes of 
death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease 
Study 2010. Lancet 2012; 380: 2095-2128. 
2. 
Global Strategy for the Diagnosis, Management and Prevention of COPD, Global Initiative for 
Chronic Obstructive Lung Disease (GOLD) 2018. Available from: http://goldcopd.org. 
3. 
Singh D, Kolsum U, Brightling CE et al. Eosinophilic inflammation in COPD: prevalence and 
clinical characteristics.  Eur Respir J 2014; 44: 1697–1700. 
4. 
Brightling CE, McKenna S, Hargadon B, et al. Sputum eosinophilia and the short term 
response to inhaled mometasone in chronic obstructive pulmonary disease. Thorax 2005; 60: 193–
198.  
5. 
Brightling CE, Monteiro W, Ward R, et al. Sputum eosinophilia and short-term response to 
prednisolone in chronic obstructive pulmonary disease: a randomised controlled trial. Lancet 2000; 
356: 1480–1485.  
6. 
Leigh R, Pizzichini MM, Morris MM, et al. Stable COPD: predicting benefit from high-dose 
inhaled corticosteroid treatment. Eur Respir J 2006; 27: 964–971.  
7. 
Pizzichini E, Pizzichini MM, Gibson P, et al. Sputum eosinophilia predicts benefit from 
prednisone in smokers with chronic obstructive bronchitis. Am J Respir Crit Care Med 1998; 158: 
1511–1517. 
8. 
Pavord ID, Chanez P, Criner GJ, et al. Mepolizumab for eosinophilic chronic obstructive 
pulmonary disease. N Engl J Med 2017; 377: 1613–1629. 
9. 
Vedel-Krogh S, Nielsen SF, Lange P, Vestbo J, Nordestgaard BG. Blood eosinophils and 
exacerbations in COPD: the Copenhagen General Populations Study. Am J Respir Crit Care Med 2016; 
193: 965-974. 
10. 
Gibson PG. Variability of blood eosinophils as a biomarker in asthma and COPD. Respirology 
2018; 23: 12 –13. 
11. 
Brusselle G, Pavord I, Landi S, et al. Blood eosinophil levels as a biomarker in COPD. Respir 
Med. 2018; 138: 21-31.   
12. 
Winkel P, Statland BE, Saunders AM, Osborn H, Kupperman H. Within-day physiologic 
variation of leukocyte types in healthy subjects as assayed by two automated leukocyte differential 
analyzers. Am J Clin Pathol. 1981; 75: 693-700. 
 

 
 
Dr. Inès Van Rossem