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Higher Monocyte Expression of TLR2 and TLR4, and Enhanced Pro-inflammatory Synergy of TLR2 with NOD2 Stimulation in Sarcoidosis

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Abstract

Introduction

Sarcoidosis is an inflammatory disease of unknown etiology. However, an infectious cause has been proposed suggesting a role for pattern-recognition receptors, such as Toll-like receptors (TLRs) and nucleotide-binding domain, leucin-rich repeat containing family proteins (NLRs), in the pathogenesis.

Objective

Our aim was to investigate whether differences in TLR2 and TLR4 expression, and the response to TLR2, TLR4, and NOD2 stimulation, are associated with sarcoidosis.

Materials and Methods

Blood mononuclear cells from sarcoidosis patients (n = 24) and healthy subjects (n = 19) were incubated with the TLR2 ligands PGN and Pam3CSK4, the TLR4 ligand LPS, the NOD2 ligand MDP, or medium alone. After 16 h, monocyte TLR2 and TLR4 expression and cytokine secretion, including TNFα, IL-1β, IL-6, IL-8, IL-10, and IL-12p70, were measured using flow cytometry and cytometric bead array.

Results

TLR2 and TLR4 expression at baseline was significantly higher in patients. Combined TLR2 and NOD2 stimulation induced a four-fold higher secretion of TNFα and a 13-fold higher secretion of IL-1β in patients. Additionally, there was a synergistic effect of TLR2 with NOD2 stimulation on induction of IL-1β in patients, whereas IL-10 was synergistically induced in healthy subjects.

Conclusion

Increased TLR expression and enhanced secretion of pro-inflammatory cytokines after combined TLR2 and NOD2 stimulation may be related to the pathogenesis of sarcoidosis.

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Acknowledgements

The authors thank Margitha Dahl, Gunnel de Forest, Heléne Blomqvist, and Berit Olsson for their technical assistance.

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Authors and Affiliations

  1. Department of Medicine, Respiratory Medicine Unit, Karolinska Institutet, Lung Research Laboratory L4:01, Karolinska University Hospital, Solna, 171 76, Stockholm, Sweden

    Maria Wikén, Johan Grunewald, Anders Eklund & Jan Wahlström

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  1. Maria Wikén
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  2. Johan Grunewald
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  3. Anders Eklund
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  4. Jan Wahlström
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Corresponding author

Correspondence to Maria Wikén.

Additional information

This study was supported by the Swedish Heart–Lung Foundation, King Oscar II Jubilee Foundation, the Swedish Research Council, the U.S. National Institutes of Health (Grant No. 1 R21 HL077579-01), the Stockholm County Council and Karolinska Institutet.

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Table EI

Characterization of Patient Subgroups (DOC 52.0 KB)

Figure E1

TLR2 (A) and TLR4 (B) expression on peripheral blood monocytes of sarcoidosis patients with Löfgren’s syndrome (white boxes) and non-Löfgren’s syndrome (grey boxes) after in vitro stimulation with ligands indicated on the x-axis. Boxes show median and interquartile range, whiskers show range. Numbers below the boxes state the number of patients and controls in each analysis. Numbers within the parenthesis state the number of Löfgren’s syndrome patients and non-Löfgren’s syndrome patients, respectively, in each analysis (DOC 32.0 KB).

Figure E2

Secretion of TNFα (A), IL-1β (B), IL-6 (C), IL-8 (D) and IL-10 (E) from human mononuclear cells of sarcoidosis patients with Löfgren’s syndrome (white boxes) and non-Löfgren’s syndrome (grey boxes) after stimulation with ligands indicated on the x-axis. Boxes show median and interquartile range, whiskers show range. Numbers below the boxes state the number of patients and controls in each analysis. Numbers within the parenthesis state the number of Löfgren’s syndrome patients and non-Löfgren’s syndrome patients, respectively, in each analysis (DOC 38.0 KB).

Figure E3

Representative histograms of flow cytometric analysis of TLR2 (a) and TLR4 (b), after stimulation with various ligands (DOC 375 KB).

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Wikén, M., Grunewald, J., Eklund, A. et al. Higher Monocyte Expression of TLR2 and TLR4, and Enhanced Pro-inflammatory Synergy of TLR2 with NOD2 Stimulation in Sarcoidosis. J Clin Immunol 29, 78–89 (2009). https://doi.org/10.1007/s10875-008-9225-0

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