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

被引：43

作者：

Hentschel, Julia ^{[1
]}

Mueller, Ulrike ^{[1
]}

Doht, Franziska ^{[1
]}

Fischer, Nele ^{[1
]}

Boeer, Klas ^{[2
]}

Sonnemann, Juergen ^{[3
]}

Hipler, Christina ^{[4
]}

Huenniger, Kerstin ^{[5
,6
]}

Kurzai, Oliver ^{[5
,6
]}

Markert, Udo R. ^{[7
]}

Mainz, Jochen G. ^{[1
]}

机构：

[1] Jena Univ Hosp, CF Ctr, D-07740 Jena, Germany

[2] Jena Univ Hosp, Inst Clin Chem & Lab Diagnost, D-07740 Jena, Germany

[3] Jena Univ Hosp, D-07740 Jena, Germany

[4] Jena Univ Hosp, Dept Dermatol, D-07740 Jena, Germany

[5] Univ Jena, Sept Res Ctr, D-07745 Jena, Germany

[6] Hans Knoell Inst, Leibniz Inst Nat Prod Res & Infect Biol, Jena, Germany

[7] Jena Univ Hosp, Dept Obstet, Placenta Lab, D-07740 Jena, Germany

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2014年 / 404卷

关键词：

Nasal lavage; Sampling technique; Centrifugation; Cytology; Cytokines; Cystic fibrosis; PSEUDOMONAS-AERUGINOSA; PARANASAL SINUSES; NEUTROPHIL GRANULES; ALLERGIC RHINITIS; CF PATIENTS; SECRETIONS; COLONIZATION; CHALLENGE; STABILITY; CHILDREN;

D O I：

10.1016/j.jim.2013.12.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Background: 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. Methods: 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. Results: NL-repetition within 0.5-4 h markedly decreased NE, IL-8 and MPO-concentrations for up to 70%. NL centrifugation up to 250 xg 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 degrees C for years led to significantly reduced mediator concentrations in NL compared to contemporary analyses, being most pronounced for IL-1 beta, IL-6 and TNF alpha. 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. Conclusions: 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 degrees C for cytokine analysis over 1-2 years with protease inhibitors supporting stability, >= 5 years storage as well as repeated freezing and thawing should be avoided. (C) 2013 Elsevier B.V. All rights reserved.

引用

页码：41 / 51

页数：11