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Murine Borrelia arthritis is highly dependent on ASC and caspase-1, but independent of NLRP3

Marije Oosting1,2, Kathrin Buffen1,2, Subbarao RK Malireddi3, Patrick Sturm4, Ineke Verschueren1,2, Marije I Koenders5, Frank L van de Veerdonk1,2, Jos WM van der Meer1,2, Mihai G Netea1,2, Thirumala-Devi Kanneganti3† and Leo AB Joosten1,2*†

1 Department of Medicine, Radboud University Nijmegen Medical Centre, Geert Grooteplein zuid 8, Nijmegen, 6525GA,The Netherlands

2 Nijmegen Institute of Infection, Inflammation and Immunity (N4i), Radboud University Nijmegen Medical Centre, Geert Grooteplein zuid 8, Nijmegen, 6525GA,The Netherlands

3 Department of Immunology, St. Jude Children's Research Hospital, 262 Danny Thomas Place, Memphis, TN 38105, USA

4 Department of Microbiology, Radboud University Nijmegen Medical Centre, Geert Grooteplein zuid 10, Nijmegen, 6525GA, The Netherlands

5 Department of Rheumatology, Radboud University Nijmegen Medical Centre, Nijmegen, Geert Grooteplein zuid 10, 6525GA, The Netherlands

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Arthritis Research & Therapy 2012, 14:R247 doi:10.1186/ar4090

Published: 13 November 2012

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Additional file 1:

Supplemental materials and methods. The missing information from the Materials and methods section.

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Additional file 2:

Figure S1. IL-1β mRNA expression is TLR2- and MyD88-dependent. (A) IL-1β mRNA expression levels (x 1000) in bone marrow-derived macrophages isolated from wild-type (WT), TLR2-, MyD88-, NOD1-, NOD2-, and RICK gene-deficient mice. mRNA expression after 24 hours of stimulation with either medium or 5 × 10^6 B. burgdorferi per mL. At least five animals per group, bars represent mean ± SEM. *P <0.05; Mann-Whitney U test, two-tailed. IL-6 (B) and TNF-α (C) mRNA expression and protein production (in ng/mL for IL-6, and pg/mL for TNF-α, respectively) by bone marrow-derived macrophages isolated from WT, TLR2-, MyD88-, NOD1-, NOD2-, and RICK gene-deficient mice. mRNA expression after 24 hours of stimulation with either medium or 5 × 10^6 B. burgdorferi per mL. At least five animals per group, bars represent mean ± SEM. MyD88, myeloid differentiation factor 88; Nod, nucleotide-binding oligomerization domain; RICK, serine-threonine protein kinase with a caspase activation and recruitment domain; TLR, Toll-like receptor.

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Additional file 3:

Figure S2. Inflammasome-independent IL-1β transcription. (A) IL-1β mRNA expression levels (× 1000) in bone marrow-derived macrophages isolated from wild-type (WT), NLRP3-, ASC-, and caspase-1 gene-deficient mice. mRNA expression after 24 hours of stimulation with either medium or 5 × 10^6 B. burgdorferi per mL. At least five animals per group, bars represent mean ± SEM. IL-6 (B) and TNF-α (C) mRNA expression and protein production (in ng/mL for IL-6, and pg/mL for TNF-α, respectively) by bone marrow-derived macrophages isolated from WT, NLRP3-, ASC-, and caspase-1 gene-deficient mice. mRNA expression after 24 hours of stimulation with either medium or 5 × 10^6 B. burgdorferi per mL. At least five animals per group, bars represent mean ± SEM. ASC, apoptosis-associated speck-like protein containing a caspase recruitment domain (CARD); NLRP3, nucleotide oligomerization domain (NOD)-like receptor P3.

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