Article

Cytokines in exhaled breath condensate of children with asthma and cystic fibrosis

Department of Paediatric Pulmonology, University Hospital Maastricht, Maastricht, The Netherlands.
Annals of allergy, asthma & immunology: official publication of the American College of Allergy, Asthma, & Immunology (Impact Factor: 2.6). 03/2006; 96(2):349-55. DOI: 10.1016/S1081-1206(10)61247-1
Source: PubMed

ABSTRACT

Inflammatory mediators in exhaled breath condensate (EBC) indicate ongoing inflammation in the lungs and might differentiate between asthma and cystic fibrosis (CF).
To evaluate the presence, concentration, and short-term variability of TH1- and TH2-mediated cytokines (interferon-gamma [IFN-gamma], tumor necrosis factor alpha [TNF-alpha], interleukin 10 [IL-10], IL-5, IL-4, and IL-2) in EBC of children with asthma or CF and in controls and to analyze the discriminating ability of inflammatory markers in EBC between children with asthma or CF and controls.
Expired air was conducted through a double-jacketed glass tube cooled by circulating ice water. In 33 asthmatic children, 12 children with CF, and 35 control children, EBC was collected during tidal breathing. Cytokines were measured using flow cytometry.
Interleukin 2, IL-4, IFN-gamma, and IL-10 were detected in 16%, 16%, 11%, and 9%, respectively, of all samples in asthma and CF. Interleukin 5 and TNF-alpha were not detected in children with CF. Cytokine concentrations did not differ significantly in children with asthma vs CF. In controls, IFN-gamma, TNF-alpha, and IL-10 were detected in 9%, 14%, and 3%, respectively; IL-2, IL-4, and IL-5 were not detected in controls.
Cytokines such as IFN-gamma, TNF-alpha, IL-10, IL-5, IL-4, and IL-2 can be detected in EBC of children with asthma or CF. However, the concentrations found are close to the detection limits of the assay used. These findings emphasize the importance of developing more sensitive techniques for the analysis of EBC and of standardizing the EBC collection method.

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Available from: Philippe P R Rosias, Jan 06, 2014
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    • "Additionally,Colombo et al.,demonstrated IL-8 correlated with clinical biomarkers of cystic fibrosis by biochip array[31]. However, more sensitive assays, such as multiple reaction monitoring (MRM), are needed as these cytokines are often near immunological reagent dependent assay's detection limits[29]. These examples emphasize the potential for using EBC as a medium to monitor lung inflammatory mediators. "
    [Show abstract] [Hide abstract] ABSTRACT: Exhaled breath condensate (EBC) has been established as a potential source of respiratory biomarkers. Compared to the numerous small molecules identified, the protein content of EBC has remained relatively unstudied due to the methodological and technical difficulties surrounding EBC analysis. In this review, we discuss the proteins identified in EBC, by mass spectrometry, focusing on the significance of those proteins identified. We will also review the limitations surrounding mass spectral EBC protein analysis emphasizing recommendations to enhance EBC protein identifications by mass spectrometry. Finally, we will provide insight into the future directions of the EBC proteomics field.
    Full-text · Article · Sep 2014
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    • "However, if the same biomarker can be detected in samples collected using a non-invasive procedure, such as saliva or urine, these could then be prioritized for development, particularly for studies intended for pediatric populations. Efforts to develop non-invasive collection methods include analysis of immune components in saliva [19,20] or induced sputum [21] and exhaled breath condensate [22]. "
    [Show abstract] [Hide abstract] ABSTRACT: The immune response plays an important role in the pathophysiology of numerous diseases including asthma, autoimmunity and cancer. Application of biomarkers of immunotoxicity in epidemiology studies and human clinical trials can improve our understanding of the mechanisms that underlie the associations between environmental exposures and development of these immune-mediated diseases. Immunological biomarkers currently used in environmental health studies include detection of key components of innate and adaptive immunity (e.g., complement, immunoglobulin and cell subsets) as well as functional responses and activation of key immune cells. The use of high-throughput assays, including flow cytometry, Luminex, and Multi-spot cytokine detection methods can further provide quantitative analysis of immune effects. Due to the complexity and redundancy of the immune response, an integrated assessment of several components of the immune responses is needed. The rapidly expanding field of immunoinformatics will also aid in the synthesis of the vast amount of data being generated. This review discusses and provides examples of how the identification and development of immunological biomarkers for use in studies of environmental exposures and immune-mediated disorders can be achieved.
    Full-text · Article · May 2011 · International Journal of Environmental Research and Public Health
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    • "Interferon-gamma (IFN-γ ) is the defining cytokine characteristic of the Th1 immune paradigm. Studies have reported detectable levels of IFN-γ in EBC with variation in disease states [5] [6] [13] [14]. IFN-γ is approximately 20 kDa in size and is produced by multiple cells in the lung. "
    [Show abstract] [Hide abstract] ABSTRACT: Manuscripts presenting cytokine (and other protein) concentrations in exhaled breath condensate (EBC) are commonly published with insufficient control experiments performed to provide confidence that the assay was accurate. Our laboratory has identified false positives in EBC with several forms of immunoassay, including ELISA and multiplex bead arrays. In this current brief investigation, we studied EBC samples from ten subjects and examined for the effect of alteration of the matrix on the assay results reported by a commercially available ELISA assay for a commonly reported EBC cytokine (interferon-gamma (IFN-γ)). The commercial immunoassay reported measurable levels of the IFN-γ signal in all samples. However, when we added to the EBC samples the proteinaceous matrix of the zero-standard from the commercial kit, we could identify no IFN-γ signal. These simple findings are consistent with our earlier observations that the EBC matrix requires adjustment for there to be any validity to immunoassays.
    Full-text · Article · Dec 2008 · Journal of Breath Research
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