Suppression of Inflammatory Arthritis by Human Gut-Derived Prevotella histicola in Humanized Mice - PubMed (original) (raw)

Suppression of Inflammatory Arthritis by Human Gut-Derived Prevotella histicola in Humanized Mice

Eric V Marietta et al. Arthritis Rheumatol. 2016 Dec.

Abstract

Objective: The gut microbiome regulates host immune homeostasis. Rheumatoid arthritis (RA) is associated with intestinal dysbiosis. This study was undertaken to test the ability of a human gut-derived commensal to modulate immune response and treat arthritis in a humanized mouse model.

Methods: We isolated a commensal bacterium, Prevotella histicola, that is native to the human gut and has systemic immune effects when administered enterally. Arthritis-susceptible HLA-DQ8 mice were immunized with type II collagen and treated with P histicola. Disease incidence, onset, and severity were monitored. Changes in gut epithelial proteins and immune response as well as systemic cellular and humoral immune responses were studied in treated mice.

Results: When treated with P histicola in prophylactic or therapeutic protocols, DQ8 mice exhibited significantly decreased incidence and severity of arthritis compared to controls. The microbial mucosal modulation of arthritis was dependent on regulation by CD103+ dendritic cells and myeloid suppressors (CD11b+Gr-1+ cells) and by generation of Treg cells (CD4+CD25+FoxP3+) in the gut, resulting in suppression of antigen-specific Th17 responses and increased transcription of interleukin-10. Treatment with P histicola led to reduced intestinal permeability by increasing expression of enzymes that produce antimicrobial peptides as well as tight junction proteins (zonula occludens 1 and occludin). However, the innate immune response via Toll-like receptor 4 (TLR-4) and TLR-9 was not affected in treated mice.

Conclusion: Our results demonstrate that enteral exposure to P histicola suppresses arthritis via mucosal regulation. P histicola is a unique commensal that can be explored as a novel therapy for RA and may have few or no side effects.

© 2016, American College of Rheumatology.

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Conflict of interest statement

No authors declare any conflicts

Figures

Figure 1

Figure 1

Oral treatment with P. histicola modulates systemic immune response in DQ8.Aβo mice. Serum levels of cytokines and chemokines were tested in naïve DQ8 transgenic mice gavaged with bacterial media alone, Sham (■) P. histicola (formula image) or P. melanogenica (formula image) every other day for two weeks. *denotes p<0.05 comparison between P. histicola treated and sham mice (N=5–7 each experimental group).

Figure 2

Figure 2

Oral treatment with P. histicola protects DQ8 mice from Collagen Induced Arthritis. P. histicola was administered to DQ8 mice in therapeutic protocol (CII+ P. histicola) (N=21) or as a prophylactic measure (P. histicola + CII), (N=12). P. melanogenica was administered as a species control for the therapeutic protocol (CII + P. melanogenica) (N=10). Other controls consisted of oral gavage with P. histicola without CII immunization (P. histicola) (N=12) and CII immunization with gavage of bacterial culture media alone (CII+ Media) (N=18) in DQ8 mice. (A) Incidence of arthritis in various experimental groups described above. Difference between CII-immunized and media versus P. histicola treated mice, incidence and severity, P<0.05 (B) Hematoxylin and eosin staining of the small intestine is shown from a representative mouse for each of the three treatment groups (P. histicola treated, sham (CII+ media) treated, and P. melanogenica treated). N=3 each group (C) Levels of serum anti-CII (IgG) antibodies before and after the administration of P. histicola using the therapeutic protocol. (D) T cell proliferative response to CII in vitro at the termination of the experiment at 12 weeks using splenocytes in mice with collagen-induced arthritis (CIA+) and without (CIA-).

Figure 3

Figure 3

Treatment with P. histicola modulates immune response in an antigen-specific manner in CIA. Splenocytes harvested from DQ8 mice in different conditions as indicated were tested for in vitro proliferative response to A) CPG and LPS, and B) CII in mice treated with P. histicola, prophylactic ( P. histicola +CII) and therapeutic (CII+ P. histicola). C) Serum cytokines produced by DQ8 transgenic mice induced for arthritis and treated with culture media (sham) or P. histicola in therapeutic protocol (P. hist.) (N=4 each group).

Figure 4

Figure 4

Anti-inflammatory effects of P. histicola treatment upon the Intestinal immune system. Fold change in cytokine transcript levels in arthritic (CIA+) and non-arthritic (CIA-) DQ8 mice treated with P. histicola treatment (therapeutic) as compared to the control group in the A) duodenum, B) jejunum, C) ileum, and D) colon. Expression levels of cytokine transcripts in duodenum, jejunum, ileum and colon of arthritic mice immunized with CII and gavaged media (Sham), and mice immunized with CII and treated with P. histicola in therapeutic protocol, arthritic (group 1) and non-arthritic (group 2) (N=3 each group). Control group was used as the reference. E) the absolute numbers of regulatory T cells (CD4+CD25+FoxP3+) producing IL-10 in the spleens of mice immunized with CII and treated with P. histicola or bacterial media alone (sham) Sham. FACS histogram of regulatory DCs, CD11c+CD103, in F) lamina propria cells and G) splenocytes. Also, shown is FACS histogram of CD4+GITR+ regulatory T cells in spleens of CII-immunized and treated or sham mice. (N=3–4 mice per group, experiment representative).

Figure 5

Figure 5

CD4+ cells from P. histicola treated mice generate lower antigen-specific T cell response compared to controls. A) In vitro T cell response to CII in splenic CD4+ cells of DQ8 mice immunized with CII and gavaged with P. histicola (P. hist.) with CD11c+ cells (DCs) from same mouse or control mouse, CII immunized mice gavaged with bacterial media alone (sham) or vice versa, B) IL-17 production in supernatant from the culture in 5A, and C) regulatory dendritic cells, CD11c+CD103+ cells and myeloid suppressors, CD11b+Gr-1+. (N=4 each group) were enumerated from splenocytes from sham and P. histicola treated mice. D) Gut permeability, done by FITC-Dextran, in naïve (N=12) and CII-immunized mice treated with P. histicola (N=12) or not (Sham) (N=8. E) Expression of ZO-1 in intestinal sections of control and P. histicola treated DQ8 mice induced for arthritis. F) Expression of Occludin in arthritic DQ8 mice (a), arthritic mice administered P. histicola (b) and naïve mice treated with P. histicola (c). Sections are shown at 60× magnification. Mean florescence intensity (MFI) of expression is depicted below.

Figure 6

Figure 6

P. histicola treatment suppresses arthritis by increasing expression of tight junction protein. A) DBA/1 mice treated with P. histicola develop milder arthritis compared to controls. DBA/1 mice were induced for arthritis and treated with P. histicola or media 7 days post immunization (N=10) and monitored for arthritis. From day 30 onwards, a significant difference in paw severity was observed between treated and control mice. P. histicola treated Caco-2 cells show increase expression of B) Zo-1 (left panel) and Occludin (right panel) and merged (middle panel) and C) fold expression of mRNA transcripts of certain cytokines in treated as compared to Caco-2 cells in media only. D) The ANI plot comparing whole-genome average nucleotide identity as calculated using the windowed blast method and maximum. E) Likelihood phylogenetic tree of the 16S rRNA gene of all the available different genomes and the P. histicola isolate from the present study.

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References

    1. Klareskog L, Padyukov L, Lorentzen J, Alfredsson L. Mechanisms of disease: Genetic susceptibility and environmental triggers in the development of rheumatoid arthritis. Nature clinical practice. 2006;2:425–33. - PubMed
    1. Gregersen PK, Silver J, Winchester RJ. The shared epitope hypothesis. An approach to understanding the molecular genetics of susceptibility to rheumatoid arthritis. Arthritis and rheumatism. 1987;30:1205–13. - PubMed
    1. Behrens M, Trejo T, Luthra H, Griffiths M, David CS, Taneja V. Mechanism by which HLA-DR4 regulates sex-bias of arthritis in humanized mice. J Autoimmun. 2010;35:1–9. - PMC - PubMed
    1. Gomez A, Luckey D, Yeoman CJ, Marietta EV, Berg Miller ME, Murray JA, et al. Loss of sex and age driven differences in the gut microbiome characterize arthritis-susceptible 0401 mice but not arthritis-resistant 0402 mice. PLoS One. 2012;7:e36095. - PMC - PubMed
    1. Toivanen P, Vaahtovuo J, Eerola E. Influence of major histocompatibility complex on bacterial composition of fecal flora. Infection and immunity. 2001;69:2372–7. - PMC - PubMed

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