The control group consisted of 19 healthy subjects (nine male, 10 female) who underwent bronchoalveolar lavage. The medical ethical committee of the St Antonius Hospital in Nieuwegein approved this study and all subjects gave formal written informed consent. All patients underwent a BAL procedure as part of the diagnostic process. The bronchoscopy with BAL was performed according to international accepted guidelines [19,20]. BAL was performed in the right middle lobe with a total volume of 200 ml saline (4 × 50 ml aliquots), which was returned in two separate fractions. The first fraction returned, after instilling 50 ml
saline, was used for microbial culture. The following three aliquots were pooled in fraction II and used Selleck FK506 for cell analysis and ELISA. Values for forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC) and diffusion capacity of the lungs for carbon monoxide (Dlco) were collected from all subjects that underwent lung function tests around the time of BAL. The parameters were expressed as a percentage of predicted values. The tests were performed according to international guidelines
. Data on blood cell counts and C-reactive protein (CRP) levels at the time of BAL as well Depsipeptide ic50 as information on mortality and history of tobacco use was collected retrospectively. MRP14 ELISA (BMA Biomedicals, Augst, Switzerland) was performed in accordance with the manufacturer’s instructions. The manufacturer has developed this ELISA in such a way that it minimizes cross-reactivity with the MRP8/14 heterodimer. The detection limit of the assay was 0·31 ng/ml. Samples that did not reach this limit were set at 50% of the detection limit. Samples equal to or lower than the negative control were set at zero. SPSS 15 (SPSS Inc., Chicago, IL, USA) and Graphpad Prism version 3 (Graphpad Software Inc., San Diego, CA, USA) were used for statistical analysis.
Analysis of variance (anova) or Student’s t-test was used to test differences in BALF MRP14 levels between patient groups. Correlations with patients’ characteristics Non-specific serine/threonine protein kinase were determined using Spearman’s rho test. Linear regression was used to test for an association with pulmonary radiographic stage in sarcoidosis patients. A P-value < 0·05 was considered significant. Control and patient characteristics are shown in Table 1. Mean BALF MRP14 levels were elevated significantly in IPF patients (P < 0·001) and sarcoidosis patients (P < 0·05) compared to controls (Fig. 1). In addition, mean BALF MRP14 levels were higher in IPF patients than in sarcoidosis patients (P < 0·01). When the sarcoidosis patients were subdivided according to chest radiographic stage, we found that the mean BALF MRP14 level was elevated significantly in stage IV sarcoidosis compared to controls (P < 0·005). When only sarcoidosis patients at presentation were included, the difference was also significant (P < 0·01).