Aberrant intestinal microbiota due to IL-1 receptor antagonist deficiency promotes IL-17- and TLR4-dependent arthritis - PubMed (original) (raw)
doi: 10.1186/s40168-017-0278-2.
Thomas H A Ederveen 1 2, Jos Boekhorst 2 3, Harm Wopereis 4 5, Jose U Scher 6, Julia Manasson 6, Sanne J C M Frambach 1, Jan Knol 4 5, Johan Garssen 4 7, Peter M van der Kraan 1, Marije I Koenders 1, Wim B van den Berg 1, Sacha A F T van Hijum 2 3, Shahla Abdollahi-Roodsaz 8 9
Affiliations
- PMID: 28645307
- PMCID: PMC5481968
- DOI: 10.1186/s40168-017-0278-2
Aberrant intestinal microbiota due to IL-1 receptor antagonist deficiency promotes IL-17- and TLR4-dependent arthritis
Rebecca Rogier et al. Microbiome. 2017.
Abstract
Background: Perturbation of commensal intestinal microbiota has been associated with several autoimmune diseases. Mice deficient in interleukin-1 receptor antagonist (Il1rn -/- mice) spontaneously develop autoimmune arthritis and are susceptible to other autoimmune diseases such as psoriasis, diabetes, and encephalomyelitis; however, the mechanisms of increased susceptibility to these autoimmune phenotypes are poorly understood. We investigated the role of interleukin-1 receptor antagonist (IL-1Ra) in regulation of commensal intestinal microbiota, and assessed the involvement of microbiota subsets and innate and adaptive mucosal immune responses that underlie the development of spontaneous arthritis in Il1rn -/- mice.
Results: Using high-throughput 16S rRNA gene sequencing, we show that IL-1Ra critically maintains the diversity and regulates the composition of intestinal microbiota in mice. IL-1Ra deficiency reduced the intestinal microbial diversity and richness, and caused specific taxonomic alterations characterized by overrepresented Helicobacter and underrepresented Ruminococcus and Prevotella. Notably, the aberrant intestinal microbiota in IL1rn -/- mice specifically potentiated IL-17 production by intestinal lamina propria (LP) lymphocytes and skewed the LP T cell balance in favor of T helper 17 (Th17) cells, an effect transferable to WT mice by fecal microbiota. Importantly, LP Th17 cell expansion and the development of spontaneous autoimmune arthritis in IL1rn -/- mice were attenuated under germ-free condition. Selective antibiotic treatment revealed that tobramycin-induced alterations of commensal intestinal microbiota, i.e., reduced Helicobacter, Flexispira, Clostridium, and Dehalobacterium, suppressed arthritis in IL1rn -/- mice. The arthritis phenotype in IL1rn -/- mice was previously shown to depend on Toll-like receptor 4 (TLR4). Using the ablation of both IL-1Ra and TLR4, we here show that the aberrations in the IL1rn -/- microbiota are partly TLR4-dependent. We further identify a role for TLR4 activation in the intestinal lamina propria production of IL-17 and cytokines involved in Th17 differentiation preceding the onset of arthritis.
Conclusions: These findings identify a critical role for IL1Ra in maintaining the natural diversity and composition of intestinal microbiota, and suggest a role for TLR4 in mucosal Th17 cell induction associated with the development of autoimmune disease in mice.
Keywords: Autoimmune arthritis; IL-1 receptor antagonist; Microbiota; T helper 17 cells; Toll-like receptors.
Figures
Fig. 1
IL-1Ra deficiency skews intestinal microbial composition and reduces its diversity in a TLR4-dependent manner. a Principal coordinates analysis (PCoA) based on an unweighted UniFrac analysis of the intestinal microbial composition where samples of mice from different cages and litters are highlighted with different colors. The position and distance of data points indicates the degree of similarity in terms of both presence and relative abundance of bacterial taxonomies. b Number of observed operational taxonomic units (OTUs). c Shannon index of microbial diversity. d Alpha diversity rarefaction curves of phylogenetic distance (PD) whole tree. e PD whole tree, bootstrapped for 2000 reads per sample, averaged of four trials. f Chao index are shown. Data (mean + SEM) represent 16S rRNA gene 454-pyrosequencing analysis of intestinal microbiota of WT (n = 9), IL1rn −/− (n = 15), IL1rn −/− Tlr2 −/− (n = 8), and IL1rn −/− Tlr4 −/− (n = 8) mice. n.s. not significant,*P ≤ 0.05 and ***P ≤ 0.001, by Mann-Whitney U test. See also Additional file 1: Figure S1 and Additional file 1: Table S1
Fig. 2
IL-1 receptor antagonist controls relative abundance of specific intestinal microbial taxa. Phylogenetic tree created by Cytoscape software showing specific changes in intestinal microbial community at different taxonomic levels induced by IL-1Ra deficiency. Nodes represent taxa, and the size of each node represents its relative abundance. The color red indicates a decrease and blue represents an increase of relative abundance in IL1rn −/− compared with WT mice. The thickness of the green border indicates the degree of the statistical significance by Mann-Whitney U test. See also Additional file 1: Table S2
Fig. 3
Il1rn −/− intestinal microbiota potentiate IL-17 production in intestinal lamina propria and joint-draining lymph nodes. a–i Production of prototypic Th1, Th17, and Th2 cell cytokines (IFNγ, IL-17A, and IL-4, respectively) by SI-LP (a–c and g–i) and draining lymph node (dLN) lymphocytes (d–f). Cells were isolated from WT and IL1rn −/− mice (a–f) or WT mice transplanted with IL1rn −/− feces and co-housed with IL1rn −/− mice for 10 days (g–i). Cells were stimulated ex vivo with PMA and ionomycin in duplicates for 5 h, and cytokines were measured by Luminex assay. Data represent mean + SEM of a representative experiment with n = 5 (a–c) and n = 3 (d–i) mice per group, each stimulated in duplicate. n.s. not significant, *P ≤ 0.05 and ***P ≤ 0.001, by Mann-Whitney U test. See also Additional file 1: Figure S2
Fig. 4
Commensal microbiota drive potentiated Th17 response and spontaneous arthritis in IL1rn −/− mice. a–d Frequency and numbers of IFNγ-producing (a, b) and IL-17-producing (c, d) CD3+CD4+ SI-LP cells. Data are pooled from three independent experiments. e Arthritis severity scores of conventional (CV, n = 12) and germ-free (GF, n = 11) IL1rn −/− mice of a representative experiment. Scale 0–2 for each hind paw. Mean + SEM is shown. f–h Production of IFNγ, IL-17, and IL-4 upon ex vivo stimulation of the spleen and lymph node cells from CV and GF mice with PMA and ionomycin for 6 h, as measured by Luminex assay. n = 3 mice per group of each stimulated in triplicate. n.s. not significant, **P ≤ 0.01 and ***P ≤ 0.001, by Mann-Whitney U test. See Additional file 1: Figure S3 for gating strategy
Fig. 5
Commensal intestinal anaerobic tobramycin-sensitive microbiota promote arthritis in IL1rn −/− mice. a Arthritis severity scores (0–2 per paw) of IL1rn −/− mice treated with a cocktail of metronidazole, neomycin, and ampicillin (ABX) for 7 days (week 5 to 6), followed by re-colonization with SFB (ABX + SFB) 1 week after ending ABX treatment (at week 7). b, c Arthritis severity scores of IL1rn −/− mice either untreated (Ctrl) or treated with the mentioned antibiotic for 8 weeks. Data show mean + SEM of n = 5–7 (a) and n = 10 (b, c) mice per group. *P ≤ 0.05, ***P ≤ 0.001, by repeated measures ANOVA with Bonferroni correction for multiple testing. d Representative images of ankle joint sections of control and tobramycin-treated mice stained with hematoxylin and eosin illustrating decreased synovial inflammation, cartilage destruction (closed arrows), and bone erosion (open arrows). Original magnification ×50 (left panels) and ×100 (right panels). e Histopathologic scores (mean + SEM) of synovial inflammation, cartilage destruction, and bone erosion in control and tobramycin-treated IL1rn −/− mice. n = 9 per group. *P ≤ 0.05, by Mann-Whitney U test
Fig. 6
Alteration of specific intestinal microbiota in IL1rn −/− mice is partly TLR4 dependent. a, b Relative abundance of Ruminococcus, Streptococcus, Xylanibacter, and Prevotella in wild-type (WT) (n = 9), IL1rn −/− (n = 15), IL1rn −/− Tlr2 −/− (n = 8), and IL1rn −/− Tlr4 −/− (n = 8) mice. Data represent relative abundances of these genera obtained by 16S rRNA gene sequencing of the fecal microbiota
Fig. 7
A significant role for TLR4 in intestinal production of cytokines involved in LP Th17 differentiation. a–c Production of IL-1β, IL-23, and IL-6 by SI-LP mononuclear cells of IL1rn −/− and IL1rn −/− Tlr4 −/− mice cultured in the presence of autoclaved IL1rn −/− complete fecal microbial antigens for 24 h. d–f Cytokine production by SI-LP lymphocytes of separately housed IL1rn −/− and IL1rn −/− Tlr4 −/− mice ex vivo stimulated with PMA and ionomycin for 5 h. g–i Cytokine production by SI-LP lymphocytes of IL1rn −/− and IL1rn −/− Tlr4 −/− mice co-housed for 10 days. Cells were stimulated as in d–f. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, by Mann-Whitney U test
References
- Carter DB, Deibel MR, Jr, Dunn CJ, Tomich CS, Laborde AL, Slightom JL, Berger AE, Bienkowski MJ, Sun FF, McEwan RN, et al. Purification, cloning, expression and biological characterization of an interleukin-1 receptor antagonist protein. Nature. 1990;344:633–638. doi: 10.1038/344633a0. - DOI - PubMed
- Nakae S, Saijo S, Horai R, Sudo K, Mori S, Iwakura Y. IL-17 production from activated T cells is required for the spontaneous development of destructive arthritis in mice deficient in IL-1 receptor antagonist. Proc Natl Acad Sci U S A. 2003;100:5986–5990. doi: 10.1073/pnas.1035999100. - DOI - PMC - PubMed
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