Protein levels of CC chemokine ligand (CCL)15, CCL16 and macrophage stimulating protein in patients with sarcoidosis

Abstract

SummaryThe objective of this study was to assess protein levels for candidate cytokines, chemokines, growth factors, matrix metalloproteinases and their inhibitors in bronchoalveolar lavage fluid (BALF) in patients with polar forms of pulmonary sarcoidosis, i.e. Löfgren's syndrome (LS) and more advanced chest X-ray (CXR) stage III disease. Twenty-four inflammatory molecules were analysed in unconcentrated BALF samples from 10 sarcoidosis patients with CXR stage III and 10 patients with LS by semiquantitative protein array. Four novel molecules [CC chemokine ligand (CCL)15, CCL16, macrophage migration inhibitory factor (MIF) and macrophage stimulating protein (MSP)], detected for the first time in association with sarcoidosis, were then quantified by enzyme-linked immunosorbent assay in a second cohort of 68 sarcoidosis patients and 17 control subjects. The protein levels of CCL15, CCL16, CCL24, CXCL8, CXCL9, CXCL10, interleukin-16, MIF, MSP and matrix metallopeptidase 1 were increased in CXR stage III patients when compared with patients with LS. CCL15 and MSP up-regulation in CXR stage III patients in comparison with LS patients and controls was confirmed by enzyme-linked immunosorbent assay. Moreover, MSP was associated with treatment requirement (P = 0·001) and CCL15 was elevated in patients with disease progression at 2-year follow-up (P = 0·016). CCL16 levels were increased in sarcoidosis versus controls (P < 0·05), but no difference was observed between patient subgroups. MIF up-regulation was not confirmed in a larger patient group. In conclusion, chemokines CCL15, CCL16 and MSP were found elevated for the first time in BALF from sarcoidosis patients; our results showed that CCL15 and MSP may affect disease course.

Chemokine receptor CXCR3 ligands in bronchoalveolar lavage fluid: associations with radiological pattern, clinical course, and prognosis in sarcoidosis

“…The increased concentrations of CXCR3 ligands in BAL fluid from patients with sarcoidosis have been reported by a number of studies 4,7-9 ; however, their results were inconsistent. For instance, Busuttil et al 4 reported elevated CXCL9 levels but did not confirm elevated CXCL10 levels in BAL fluid in this patient group, while Arakelyan et al 17 did not find significant differences in the BAL fluid levels of CXCL9 and CXCL10 between the sarcoidosis and control groups, which is in line with our study. These discrepancies may be explained by the polymorphic and highly variable clinical course of the disease and different patient selection criteria.…”

T-helper cell type-1 transcription factor T-bet is upregulated in pulmonary sarcoidosis

“…Of the studied chemokine receptors, we observed a strong correlation between T-bet and CXCR6, a receptor co-expressed on CXCR3 T-cells in sarcoidosis [4]. Furthermore, a strong correlation was observed between T-bet and CCL5 and CXCL10, two chemokines that play an important role in the recruitment of T-cells to the sarcoid lungs [4,[15][16][17]19]. Additionally, we observed a strong association between T-bet and IL-2R/IL-15Rb (CD122) in our sarcoidosis patients; this association has been already reported in animal cell cultures [5,7].…”

“…Determination of cell-associated T-bet protein by immunocytochemistry T-bet protein was detected on the acetone-chloroform fixed cytocentrifuge preparations of BAL cells using the streptavidin-biotin/horseradish peroxidase method using an anti-T-bet antibody (clone 525803, dilution 1:50; R&D Systems Inc.) and a negative control mouse IgG1 (clone DAK-GO1, dilution 1:50; Dako) as described previously [16]. …”

“…To address the role for T-bet as a plausible regulator of immune mediators in sarcoidosis, we studied its relationship with cytokines and their receptors that have been previously implicated in the pathogenesis of sarcoidosis. Of these ''sarcoidosis-associated'' genes we mainly focused on chemokines directing the trafficking of activated T-cells into sarcoid lungs, such as CCL2, CCL5, CXCL9, CXCL10 and CXCL11 [14][15][16][17][18][19], and chemokine receptors, CC chemokine ligand (CCR)5 [10] and CXCR6 [4] preferentially expressed on activated lymphocytes.…”

“…Secondly, we aimed to address the question as to whether there is evidence supporting the theory of T-bet modulation of CXCR3 and IFN-c gene expression in sarcoid lungs by performing correlation analyses of their mRNA expression profiles in BAL cells. Finally, we investigated the relationship of T-bet with a range of sarcoidosis-associated genes, namely: 1) ligands of CXCR3 receptor (CXCL9, CXCL10 and CXCL11) [14][15][16][17]; 2) chemokines CCL2 and CCL5 [18,19]; 3) chemokine receptors CCR2, CCR5 and CXCR6 [4,10,20,21]; and 4) b-chain subunit of IL-2R and IL-15R receptors [10,22]. Herein, we report upregulation of T-bet in sarcoidosis and its relationship with some sarcoid-associated molecules.…”