Influences of nasal lavage collection-, processing- and storage methods on inflammatory markers — Evaluation of a method for non-invasive sampling of epithelial lining fluid in cystic fibrosis and other respiratory diseases

Non-invasive sampling of airway epithelial-lining-fluid by nasal lavage (NL) is an emerging method to monitor allergy, infection and inflammation in patients with respiratory diseases. However, the influences of collection-, processing- and storage-methods have not been sufficiently evaluated and st...

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Bibliographic Details
Published in:Journal of immunological methods 2014-02, Vol.404, p.41-51
Main Authors: Hentschel, Julia, Müller, Ulrike, Doht, Franziska, Fischer, Nele, Böer, Klas, Sonnemann, Jürgen, Hipler, Christina, Hünniger, Kerstin, Kurzai, Oliver, Markert, Udo R., Mainz, Jochen G.
Format: Article
Language:English
Subjects:
Adolescent
Adult
Aged
Biomarkers - analysis
Case-Control Studies
Centrifugation
Centrifugation - standards
Child
Child, Preschool
Cystic fibrosis
Cystic Fibrosis - diagnosis
Cystic Fibrosis - immunology
Cystic Fibrosis - metabolism
Cytokines
Cytology
Enzyme-Linked Immunosorbent Assay
Female
Freezing
Humans
Inflammation
Interleukin-1 - analysis
Interleukin-6 - analysis
Interleukin-8 - analysis
Leukocyte Elastase - analysis
Male
Middle Aged
Nasal lavage
Nasal Lavage Fluid - chemistry
Peroxidase - analysis
Protease Inhibitors - chemistry
Sampling technique
Specimen Handling - standards
Tumor Necrosis Factor-alpha - analysis
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Summary:Non-invasive sampling of airway epithelial-lining-fluid by nasal lavage (NL) is an emerging method to monitor allergy, infection and inflammation in patients with respiratory diseases. However, the influences of collection-, processing- and storage-methods have not been sufficiently evaluated and standardized. Influences of repeated NL, centrifugation setups, repeated freezing and thawing, and protease inhibitors on mediator concentration were evaluated in healthy controls and CF patients, which serve as a model for chronic bacterial infection and inflammation. Polymorphonuclear leukocyte elastase (NE)/myeloperoxidase (MPO)/interleukin (IL)-1/IL-6/IL-8 and tumour necrosis factor alpha (TNF) concentrations were measured using ELISA and Multiplex Bead-Arrays. NL-repetition within 0.5–4h markedly decreased NE, IL-8 and MPO-concentrations for up to 70%. NL centrifugation up to 250×g for cellular differentiation did not significantly influence mediator concentration in native and processed NL fluid. NL freezing and thawing markedly decreased IL-8 and MPO concentrations by up to 50% while NE remained stable. In contrast to preceding reports, storing at −70°C for ≥5years led to significantly reduced mediator concentrations in NL compared to contemporary analyses, being most pronounced for IL-1β, IL-6 and TNFa. Storing of samples in the presence of protease inhibitors led to an increase in marker concentration for IL-8 (+27%) and MPO (+15%) even after one year of storage. NL is an easy and robust technique for inflammation monitoring of the upper airways. For the first time we have shown that diagnostic NL should be performed only once daily to get comparable results. Whereas NL-fluid can be stored unprocessed at −70°C for cytokine analysis over 1–2years with protease inhibitors supporting stability, ≥5years storage as well as repeated freezing and thawing should be avoided. •nasal lavage is an easy and robust technique for airway inflammation monitoring.•repetition of NL within four hours reduces MPO concentration about 70%.•Diagnostic NL should be performed only once daily for assessment of inflammation.•NL-fluid can be stored unprocessed at -70°C for cytokine analysis over 1-2years.•protease inhibitors support stability.•repeated freezing and thawing should be avoided.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2013.12.003