Limits...
Local and Systemic IKKε and NF-κB Signaling Associated with Sjögren's Syndrome Immunopathogenesis.

Chen W, Lin J, Cao H, Xu D, Xu B, Xu L, Yue L, Sun C, Wu G, Qian W - J Immunol Res (2015)

Bottom Line: In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative.Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity.These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China.

ABSTRACT
The activated NF-κB signaling pathway plays an important role in pathogenesis of primary Sjögren's syndrome (pSS). The inhibitor of κB (IκB) kinase (IKK) family such as IKKα, IKKβ, IKKγ, and IKKε, is required for this signaling. Our aim was to investigate the role of IKKα/β/γ/ε in patients with untreated pSS. In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative. And pIKKε positively related to expression of p-p65. Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity. Meanwhile, in peripheral blood mononuclear cells from pSS patients, pIKKε, total IKKε, pIKKα/β, and p-p65 were significantly increased by western blot, compared to healthy controls. However, there was no difference in IKKγ and IκBα between pSS patients and healthy individuals. These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

No MeSH data available.


Related in: MedlinePlus

Protein expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were significantly increased in PBMC from patients with pSS, compared to healthy controls. (a) Phosphorylated and total protein expression of IKKε, IKKα/β, IKKγ, IκBα, and NF-κB p65 in PBMC from two healthy controls (H) and two representative pSS patients was shown by western blot. (b) The relative expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were higher in pSS (n = 33), compared to healthy controls (n = 26). β-actin was used as endogenous control, and relative expression of each protein is shown as protein/β-actin ratio. ∗P < 0.05;  ∗∗∗P < 0.001. All data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4563092&req=5

fig2: Protein expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were significantly increased in PBMC from patients with pSS, compared to healthy controls. (a) Phosphorylated and total protein expression of IKKε, IKKα/β, IKKγ, IκBα, and NF-κB p65 in PBMC from two healthy controls (H) and two representative pSS patients was shown by western blot. (b) The relative expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were higher in pSS (n = 33), compared to healthy controls (n = 26). β-actin was used as endogenous control, and relative expression of each protein is shown as protein/β-actin ratio. ∗P < 0.05;  ∗∗∗P < 0.001. All data are representative of three independent experiments.

Mentions: To learn more about the activity of the proinflammatory NF-κB pathway in cases of pSS, we also looked at peripheral blood mononuclear cells to see if we could characterize NF-κB activation in the white blood cells that mediate much of the autoimmunity. Using western blot of cell lysates, we saw that protein expression of pIKKε, total IKKε, and pNF-κB p65 was significantly increased in PBMC from patients with pSS when compared to healthy controls (Figures 2(a) and 2(b)). Phosphorylation of IKKα/β was also enhanced in pSS (Figures 2(a) and 2(b)) while there was no difference in protein expression of total IKKα/β, pIKKγ, total IKKγ, pIκBα, total IκBα, or NF-κB p65 between pSS patients and healthy individuals (Figure 2(a), Supplementary Figure S5A, 5B). Untreated Jurkat T leukemia cells served as a negative control for pIKKε (Supplementary Figure S3), as reported previously [8]. The sizes of the proteins in question can be seen on the uncropped blots shown in Supplementary Figure S4.


Local and Systemic IKKε and NF-κB Signaling Associated with Sjögren's Syndrome Immunopathogenesis.

Chen W, Lin J, Cao H, Xu D, Xu B, Xu L, Yue L, Sun C, Wu G, Qian W - J Immunol Res (2015)

Protein expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were significantly increased in PBMC from patients with pSS, compared to healthy controls. (a) Phosphorylated and total protein expression of IKKε, IKKα/β, IKKγ, IκBα, and NF-κB p65 in PBMC from two healthy controls (H) and two representative pSS patients was shown by western blot. (b) The relative expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were higher in pSS (n = 33), compared to healthy controls (n = 26). β-actin was used as endogenous control, and relative expression of each protein is shown as protein/β-actin ratio. ∗P < 0.05;  ∗∗∗P < 0.001. All data are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4563092&req=5

fig2: Protein expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were significantly increased in PBMC from patients with pSS, compared to healthy controls. (a) Phosphorylated and total protein expression of IKKε, IKKα/β, IKKγ, IκBα, and NF-κB p65 in PBMC from two healthy controls (H) and two representative pSS patients was shown by western blot. (b) The relative expression of pIKKε, total IKKε, pIKKα/β, and pNF-κB p65 were higher in pSS (n = 33), compared to healthy controls (n = 26). β-actin was used as endogenous control, and relative expression of each protein is shown as protein/β-actin ratio. ∗P < 0.05;  ∗∗∗P < 0.001. All data are representative of three independent experiments.
Mentions: To learn more about the activity of the proinflammatory NF-κB pathway in cases of pSS, we also looked at peripheral blood mononuclear cells to see if we could characterize NF-κB activation in the white blood cells that mediate much of the autoimmunity. Using western blot of cell lysates, we saw that protein expression of pIKKε, total IKKε, and pNF-κB p65 was significantly increased in PBMC from patients with pSS when compared to healthy controls (Figures 2(a) and 2(b)). Phosphorylation of IKKα/β was also enhanced in pSS (Figures 2(a) and 2(b)) while there was no difference in protein expression of total IKKα/β, pIKKγ, total IKKγ, pIκBα, total IκBα, or NF-κB p65 between pSS patients and healthy individuals (Figure 2(a), Supplementary Figure S5A, 5B). Untreated Jurkat T leukemia cells served as a negative control for pIKKε (Supplementary Figure S3), as reported previously [8]. The sizes of the proteins in question can be seen on the uncropped blots shown in Supplementary Figure S4.

Bottom Line: In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative.Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity.These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, China.

ABSTRACT
The activated NF-κB signaling pathway plays an important role in pathogenesis of primary Sjögren's syndrome (pSS). The inhibitor of κB (IκB) kinase (IKK) family such as IKKα, IKKβ, IKKγ, and IKKε, is required for this signaling. Our aim was to investigate the role of IKKα/β/γ/ε in patients with untreated pSS. In minor salivary glands from pSS patients, phosphorylated IKKε (pIKKε), pIκBα, and pNF-κB p65 (p-p65) were highly expressed in ductal epithelium and infiltrating mononuclear cells by immunohistochemistry, compared to healthy individuals. pIKKα/β and pIKKγ were both negative. And pIKKε positively related to expression of p-p65. Furthermore, pIKKε and p-p65 expression significantly correlated with biopsy focus score and overall disease activity. Meanwhile, in peripheral blood mononuclear cells from pSS patients, pIKKε, total IKKε, pIKKα/β, and p-p65 were significantly increased by western blot, compared to healthy controls. However, there was no difference in IKKγ and IκBα between pSS patients and healthy individuals. These results demonstrated an abnormality of IKKε, IκBα, and NF-κB in pSS, suggesting a potential target of treatment for pSS based on the downregulation of IKKε expression and deregulation of NF-κB pathway.

No MeSH data available.


Related in: MedlinePlus