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Antagonistic Interplay between MicroRNA-155 and IL-10 during Lyme Carditis and Arthritis.

Lochhead RB, Zachary JF, Dalla Rosa L, Ma Y, Weis JH, O'Connell RM, Weis JJ - PLoS ONE (2015)

Bottom Line: Using B. burgdorferi-infected B6, Mir155-/-, Il10-/-, and Mir155-/- Il10-/- double-knockout (DKO) mice, we found that anti-inflammatory IL-10 and pro-inflammatory miR-155 have opposite and somewhat compensatory effects on myeloid cell activity, cytokine production, and antibody response.Both IL-10 and miR-155 were required for suppression of Lyme carditis.These results show that antagonistic interplay between IL-10 and miR-155 is required to balance host defense and immune activation in vivo, and this balance is particularly important for suppression of Lyme carditis.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Immunology, Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.

ABSTRACT
MicroRNA-155 has been shown to play a role in immune activation and inflammation, and is suppressed by IL-10, an important anti-inflammatory cytokine. The established involvement of IL-10 in the murine model of Borrelia burgdorferi-induced Lyme arthritis and carditis allowed us to assess the interplay between IL-10 and miR-155 in vivo. As reported previously, Mir155 was highly upregulated in joints from infected severely arthritic B6 Il10-/- mice, but not in mildly arthritic B6 mice. In infected hearts, Mir155 was upregulated in both strains, suggesting a role of miR-155 in Lyme carditis. Using B. burgdorferi-infected B6, Mir155-/-, Il10-/-, and Mir155-/- Il10-/- double-knockout (DKO) mice, we found that anti-inflammatory IL-10 and pro-inflammatory miR-155 have opposite and somewhat compensatory effects on myeloid cell activity, cytokine production, and antibody response. Both IL-10 and miR-155 were required for suppression of Lyme carditis. Infected Mir155-/- mice developed moderate/severe carditis, had higher B. burgdorferi numbers, and had reduced Th1 cytokine expression in hearts. In contrast, while Il10-/- and DKO mice also developed severe carditis, hearts had reduced bacterial numbers and elevated Th1 and innate cytokine expression. Surprisingly, miR-155 had little effect on Lyme arthritis. These results show that antagonistic interplay between IL-10 and miR-155 is required to balance host defense and immune activation in vivo, and this balance is particularly important for suppression of Lyme carditis. These results also highlight tissue-specific differences in Lyme arthritis and carditis pathogenesis, and reveal the importance of IL-10-mediated regulation of miR-155 in maintaining healthy immunity.

No MeSH data available.


Related in: MedlinePlus

Activation of macrophages is modulated by IL-10/miR-155 regulatory network.Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and cells were stimulated with media alone or with B. burgdorferi. A, Expression of Mir155 in BMDMs stimulated with B. burgdorferi for 24 hours by quantitative real-time PCR (qRT-PCR). B, Peritoneal macrophages were incubated with GFP-expressing B. burgdorferi for 1 or 2 hours, and cells containing intracellular bacteria (GFP-positive) were determined by flow cytometric analysis. C,Cxcl9 and Cxcl10 expression in BMDMs stimulated with B. burgdorferi for 24 hours, by qRT-PCR. D, Cytokine levels in culture supernatant from BMDMs stimulated by B. burgdorferi for 24 hours, measured by ELISA. Statistically significant differences between strains were determined by ANOVA (Tukey’s post-hoc), and are indicated (* p<0.05), with adjusted p-values included for trends not achieving statistical significance (p<0.1). Results are from one experiment (n = 3 triplicates per mouse strain).
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pone.0135142.g001: Activation of macrophages is modulated by IL-10/miR-155 regulatory network.Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and cells were stimulated with media alone or with B. burgdorferi. A, Expression of Mir155 in BMDMs stimulated with B. burgdorferi for 24 hours by quantitative real-time PCR (qRT-PCR). B, Peritoneal macrophages were incubated with GFP-expressing B. burgdorferi for 1 or 2 hours, and cells containing intracellular bacteria (GFP-positive) were determined by flow cytometric analysis. C,Cxcl9 and Cxcl10 expression in BMDMs stimulated with B. burgdorferi for 24 hours, by qRT-PCR. D, Cytokine levels in culture supernatant from BMDMs stimulated by B. burgdorferi for 24 hours, measured by ELISA. Statistically significant differences between strains were determined by ANOVA (Tukey’s post-hoc), and are indicated (* p<0.05), with adjusted p-values included for trends not achieving statistical significance (p<0.1). Results are from one experiment (n = 3 triplicates per mouse strain).

Mentions: Bone marrow-derived macrophages (BMDMs) were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and stimulated in vitro with B. burgdorferi (Fig 1). As expected, Mir155 was not detectible by qRT-PCR in Mir155-/- and DKO BMDMs, and cells lacking IL-10 had elevated levels of Mir155 upon stimulation, compared to B6 (Fig 1A), which was shown previously [22].


Antagonistic Interplay between MicroRNA-155 and IL-10 during Lyme Carditis and Arthritis.

Lochhead RB, Zachary JF, Dalla Rosa L, Ma Y, Weis JH, O'Connell RM, Weis JJ - PLoS ONE (2015)

Activation of macrophages is modulated by IL-10/miR-155 regulatory network.Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and cells were stimulated with media alone or with B. burgdorferi. A, Expression of Mir155 in BMDMs stimulated with B. burgdorferi for 24 hours by quantitative real-time PCR (qRT-PCR). B, Peritoneal macrophages were incubated with GFP-expressing B. burgdorferi for 1 or 2 hours, and cells containing intracellular bacteria (GFP-positive) were determined by flow cytometric analysis. C,Cxcl9 and Cxcl10 expression in BMDMs stimulated with B. burgdorferi for 24 hours, by qRT-PCR. D, Cytokine levels in culture supernatant from BMDMs stimulated by B. burgdorferi for 24 hours, measured by ELISA. Statistically significant differences between strains were determined by ANOVA (Tukey’s post-hoc), and are indicated (* p<0.05), with adjusted p-values included for trends not achieving statistical significance (p<0.1). Results are from one experiment (n = 3 triplicates per mouse strain).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4529177&req=5

pone.0135142.g001: Activation of macrophages is modulated by IL-10/miR-155 regulatory network.Bone marrow-derived macrophages (BMDMs) and peritoneal macrophages were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and cells were stimulated with media alone or with B. burgdorferi. A, Expression of Mir155 in BMDMs stimulated with B. burgdorferi for 24 hours by quantitative real-time PCR (qRT-PCR). B, Peritoneal macrophages were incubated with GFP-expressing B. burgdorferi for 1 or 2 hours, and cells containing intracellular bacteria (GFP-positive) were determined by flow cytometric analysis. C,Cxcl9 and Cxcl10 expression in BMDMs stimulated with B. burgdorferi for 24 hours, by qRT-PCR. D, Cytokine levels in culture supernatant from BMDMs stimulated by B. burgdorferi for 24 hours, measured by ELISA. Statistically significant differences between strains were determined by ANOVA (Tukey’s post-hoc), and are indicated (* p<0.05), with adjusted p-values included for trends not achieving statistical significance (p<0.1). Results are from one experiment (n = 3 triplicates per mouse strain).
Mentions: Bone marrow-derived macrophages (BMDMs) were isolated from B6, Mir155-/-, Il10-/-, and DKO mice, and stimulated in vitro with B. burgdorferi (Fig 1). As expected, Mir155 was not detectible by qRT-PCR in Mir155-/- and DKO BMDMs, and cells lacking IL-10 had elevated levels of Mir155 upon stimulation, compared to B6 (Fig 1A), which was shown previously [22].

Bottom Line: Using B. burgdorferi-infected B6, Mir155-/-, Il10-/-, and Mir155-/- Il10-/- double-knockout (DKO) mice, we found that anti-inflammatory IL-10 and pro-inflammatory miR-155 have opposite and somewhat compensatory effects on myeloid cell activity, cytokine production, and antibody response.Both IL-10 and miR-155 were required for suppression of Lyme carditis.These results show that antagonistic interplay between IL-10 and miR-155 is required to balance host defense and immune activation in vivo, and this balance is particularly important for suppression of Lyme carditis.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Immunology, Department of Pathology, University of Utah, Salt Lake City, Utah, United States of America.

ABSTRACT
MicroRNA-155 has been shown to play a role in immune activation and inflammation, and is suppressed by IL-10, an important anti-inflammatory cytokine. The established involvement of IL-10 in the murine model of Borrelia burgdorferi-induced Lyme arthritis and carditis allowed us to assess the interplay between IL-10 and miR-155 in vivo. As reported previously, Mir155 was highly upregulated in joints from infected severely arthritic B6 Il10-/- mice, but not in mildly arthritic B6 mice. In infected hearts, Mir155 was upregulated in both strains, suggesting a role of miR-155 in Lyme carditis. Using B. burgdorferi-infected B6, Mir155-/-, Il10-/-, and Mir155-/- Il10-/- double-knockout (DKO) mice, we found that anti-inflammatory IL-10 and pro-inflammatory miR-155 have opposite and somewhat compensatory effects on myeloid cell activity, cytokine production, and antibody response. Both IL-10 and miR-155 were required for suppression of Lyme carditis. Infected Mir155-/- mice developed moderate/severe carditis, had higher B. burgdorferi numbers, and had reduced Th1 cytokine expression in hearts. In contrast, while Il10-/- and DKO mice also developed severe carditis, hearts had reduced bacterial numbers and elevated Th1 and innate cytokine expression. Surprisingly, miR-155 had little effect on Lyme arthritis. These results show that antagonistic interplay between IL-10 and miR-155 is required to balance host defense and immune activation in vivo, and this balance is particularly important for suppression of Lyme carditis. These results also highlight tissue-specific differences in Lyme arthritis and carditis pathogenesis, and reveal the importance of IL-10-mediated regulation of miR-155 in maintaining healthy immunity.

No MeSH data available.


Related in: MedlinePlus