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Non-classical MHC I-E negatively regulates macrophage activation and Th17 cell development in NOD mice.

Yang C, Guo N, Liu J, Yang J, Zhu K, Xiao H, Leng Q - Sci Rep (2015)

Bottom Line: The reduction in Th17 cells was associated with both attenuated IL-6 production and decreased activation of macrophages.In contrast to classical MHC II molecule, this non-classical MHC II molecule negatively regulates the inflammatory responses of macrophages.Altogether, our study reveals a novel regulatory role of I-E molecules in modulating inflammatory immune responses.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai, China.

ABSTRACT
Transgenic expression of I-E molecules prevents diabetes in NOD mice. So far, the precise role of these non-classical MHC II molecules remains elusive. Here, we showed that transgenic expression of I-E(k) alpha 16 molecule in NOD mice selectively reduced Th17 cells in the thymus and pancreatic draining lymph nodes. The reduction in Th17 cells was associated with both attenuated IL-6 production and decreased activation of macrophages. Mechanistically, transgenic expression of the I-E molecule diminished expression of intracellular classical MHC II molecule and led to impaired TLR4-mediated signaling. In contrast to classical MHC II molecule, this non-classical MHC II molecule negatively regulates the inflammatory responses of macrophages. Altogether, our study reveals a novel regulatory role of I-E molecules in modulating inflammatory immune responses.

No MeSH data available.


Related in: MedlinePlus

I-E molecule attenuates macrophage activation and proinflammatory cytokine production.(A) qRT-PCR analysis of the cytokine profile in the thymi from EA16 and WT NOD mice. Expression levels of cytokines were normalized to GAPDH. (B) Representative images (n = 5) for the histological analysis of thymus tissue from EA16 (below) and WT (above) NOD mice. The samples were processed for both immunohistochemical staining with anti-IL-6 antibody and haematoxylin counterstaining to detect IL-6 producing cells. The medulla areas that had high staining of IL-6 were further magnified. (C) Mean fluorescence intensity (MFI) of transgenic I-E expression on CD11b+ macrophages in the spleens and thymi from EA16 and WT NOD mice. CD11b+ macrophages were gated on the CD3e-B220- CD11b+ population. (D) CD80 and CD86 expression levels on CD11b+ macrophages in the spleens and thymi from EA16 (solid line) and WT (dashed line) NOD mice were detected by flow cytometry. (E) MFI of CD86 expression from (D). Data (mean ± SEM) are representative of two independent experiments (A,B) (n = 6 per genotype) or three independent experiments (C–E) with similar results (n = 3 per genotype). *P < 0.05.
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f3: I-E molecule attenuates macrophage activation and proinflammatory cytokine production.(A) qRT-PCR analysis of the cytokine profile in the thymi from EA16 and WT NOD mice. Expression levels of cytokines were normalized to GAPDH. (B) Representative images (n = 5) for the histological analysis of thymus tissue from EA16 (below) and WT (above) NOD mice. The samples were processed for both immunohistochemical staining with anti-IL-6 antibody and haematoxylin counterstaining to detect IL-6 producing cells. The medulla areas that had high staining of IL-6 were further magnified. (C) Mean fluorescence intensity (MFI) of transgenic I-E expression on CD11b+ macrophages in the spleens and thymi from EA16 and WT NOD mice. CD11b+ macrophages were gated on the CD3e-B220- CD11b+ population. (D) CD80 and CD86 expression levels on CD11b+ macrophages in the spleens and thymi from EA16 (solid line) and WT (dashed line) NOD mice were detected by flow cytometry. (E) MFI of CD86 expression from (D). Data (mean ± SEM) are representative of two independent experiments (A,B) (n = 6 per genotype) or three independent experiments (C–E) with similar results (n = 3 per genotype). *P < 0.05.

Mentions: To characterize the cytokine milieu that is associated with the decreased level of Th17 cells in EA16 mice, we examined the expression levels of cytokines that are crucial for Th17 development by quantitative reverse transcription-PCR (qRT-PCR). We found that the expression of cytokines, including IL-1β, IL-2, IL-4, IL-10, IL-21, TGF-β and TNF-α, in the thymus of EA16 mice was not significantly different from that of WT NOD mice (Fig. 3A and Fig. S3). In contrast, IL-6 expression levels in EA16 mice dropped to 22% of that in WT NOD mice (Fig. 3A). ELISA assay and immunohistochemical (IHC) staining also confirmed that EA16 mice indeed had less IL-6 expression in thymus than WT NOD mice (Fig. 3B and Fig. S3).


Non-classical MHC I-E negatively regulates macrophage activation and Th17 cell development in NOD mice.

Yang C, Guo N, Liu J, Yang J, Zhu K, Xiao H, Leng Q - Sci Rep (2015)

I-E molecule attenuates macrophage activation and proinflammatory cytokine production.(A) qRT-PCR analysis of the cytokine profile in the thymi from EA16 and WT NOD mice. Expression levels of cytokines were normalized to GAPDH. (B) Representative images (n = 5) for the histological analysis of thymus tissue from EA16 (below) and WT (above) NOD mice. The samples were processed for both immunohistochemical staining with anti-IL-6 antibody and haematoxylin counterstaining to detect IL-6 producing cells. The medulla areas that had high staining of IL-6 were further magnified. (C) Mean fluorescence intensity (MFI) of transgenic I-E expression on CD11b+ macrophages in the spleens and thymi from EA16 and WT NOD mice. CD11b+ macrophages were gated on the CD3e-B220- CD11b+ population. (D) CD80 and CD86 expression levels on CD11b+ macrophages in the spleens and thymi from EA16 (solid line) and WT (dashed line) NOD mice were detected by flow cytometry. (E) MFI of CD86 expression from (D). Data (mean ± SEM) are representative of two independent experiments (A,B) (n = 6 per genotype) or three independent experiments (C–E) with similar results (n = 3 per genotype). *P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4528198&req=5

f3: I-E molecule attenuates macrophage activation and proinflammatory cytokine production.(A) qRT-PCR analysis of the cytokine profile in the thymi from EA16 and WT NOD mice. Expression levels of cytokines were normalized to GAPDH. (B) Representative images (n = 5) for the histological analysis of thymus tissue from EA16 (below) and WT (above) NOD mice. The samples were processed for both immunohistochemical staining with anti-IL-6 antibody and haematoxylin counterstaining to detect IL-6 producing cells. The medulla areas that had high staining of IL-6 were further magnified. (C) Mean fluorescence intensity (MFI) of transgenic I-E expression on CD11b+ macrophages in the spleens and thymi from EA16 and WT NOD mice. CD11b+ macrophages were gated on the CD3e-B220- CD11b+ population. (D) CD80 and CD86 expression levels on CD11b+ macrophages in the spleens and thymi from EA16 (solid line) and WT (dashed line) NOD mice were detected by flow cytometry. (E) MFI of CD86 expression from (D). Data (mean ± SEM) are representative of two independent experiments (A,B) (n = 6 per genotype) or three independent experiments (C–E) with similar results (n = 3 per genotype). *P < 0.05.
Mentions: To characterize the cytokine milieu that is associated with the decreased level of Th17 cells in EA16 mice, we examined the expression levels of cytokines that are crucial for Th17 development by quantitative reverse transcription-PCR (qRT-PCR). We found that the expression of cytokines, including IL-1β, IL-2, IL-4, IL-10, IL-21, TGF-β and TNF-α, in the thymus of EA16 mice was not significantly different from that of WT NOD mice (Fig. 3A and Fig. S3). In contrast, IL-6 expression levels in EA16 mice dropped to 22% of that in WT NOD mice (Fig. 3A). ELISA assay and immunohistochemical (IHC) staining also confirmed that EA16 mice indeed had less IL-6 expression in thymus than WT NOD mice (Fig. 3B and Fig. S3).

Bottom Line: The reduction in Th17 cells was associated with both attenuated IL-6 production and decreased activation of macrophages.In contrast to classical MHC II molecule, this non-classical MHC II molecule negatively regulates the inflammatory responses of macrophages.Altogether, our study reveals a novel regulatory role of I-E molecules in modulating inflammatory immune responses.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai, China.

ABSTRACT
Transgenic expression of I-E molecules prevents diabetes in NOD mice. So far, the precise role of these non-classical MHC II molecules remains elusive. Here, we showed that transgenic expression of I-E(k) alpha 16 molecule in NOD mice selectively reduced Th17 cells in the thymus and pancreatic draining lymph nodes. The reduction in Th17 cells was associated with both attenuated IL-6 production and decreased activation of macrophages. Mechanistically, transgenic expression of the I-E molecule diminished expression of intracellular classical MHC II molecule and led to impaired TLR4-mediated signaling. In contrast to classical MHC II molecule, this non-classical MHC II molecule negatively regulates the inflammatory responses of macrophages. Altogether, our study reveals a novel regulatory role of I-E molecules in modulating inflammatory immune responses.

No MeSH data available.


Related in: MedlinePlus