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Inflammation-mediated upregulation of centrosomal protein 110, a negative modulator of ciliogenesis, in patients with chronic rhinosinusitis  Yinyan Lai,

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Presentation on theme: "Inflammation-mediated upregulation of centrosomal protein 110, a negative modulator of ciliogenesis, in patients with chronic rhinosinusitis  Yinyan Lai,"— Presentation transcript:

1 Inflammation-mediated upregulation of centrosomal protein 110, a negative modulator of ciliogenesis, in patients with chronic rhinosinusitis  Yinyan Lai, MD, Bei Chen, MD, Jianbo Shi, MD, James N. Palmer, MD, David W. Kennedy, MD, Noam A. Cohen, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 128, Issue 6, Pages e1 (December 2011) DOI: /j.jaci Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Sinonasal mucosa from patients with CRS with and without nasal polyps is poorly ciliated and overexpresses hCp110. A, Control mucosa evinces near-complete coverage by cilia, whereas mucosa from patients with CRS has significantly less ciliated coverage. B, hCp110 mRNA copy number is increased in mucosa from patients with CRS. ★P ≤ .05. C, hCp110 protein expression is increased in mucosa from patients with CRS. CRSsNP, CRS without nasal polyps; CRSwNP, CRS with nasal polyps. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Expression of mCp110 in MSEC ALI cultures inversely correlates to ciliogenesis. A, Ciliogenesis occurs over 14 days in murine sinonasal ALI cultures, as demonstrated by type IV β-tubulin staining. B, mCp110 mRNA copy number decreases during ciliogenesis (n = 4). ★★★P ≤ C, mCp110 protein expression decreases as ciliogenesis occurs (n = 4 for each time point). Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Expression of hCp110 in cultures established from control subjects and patients with CRS with nasal polyps (CRSwNP). A, Cultures established from CRSwNP samples (n = 4) demonstrate both initial and persistent increased expression of Cp110 compared with that seen in cultures derived from patients without CRS (n = 4 patients) ★P ≤ .05. B, Type IV β-tubulin staining of ALI cultures derived from control subjects and patients with CRSwNP. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Effect of CRS-associated inflammatory cytokines on ciliogenesis in HSEC and MSEC cultures. A and B, Murine septal ALI (Fig 4, A) or human sinonasal ALI (Fig 4, B) cultures derived from control subjects were incubated throughout the 14 days after air exposure with combinations of proinflammatory cytokines (TNF-α [10 ng/mL] and IL-8 or KC [5 ng/mL] or IL-13 [100 ng/mL] and IL-6 [100 ng/mL]; n = 3). Graphic representation of surface area covered by cilia demonstrates cytokine-mediated inhibition of ciliogenesis. We show the mean percentage from 10 independent experiments. Vertical bars indicate SDs. ★★★P ≤ .001. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Expression of Cp110 mRNA in MSEC and HSEC ALI cultures with stimulation of cytokine mixtures. On the 14th day, both MSEC (n = 3; A) and HSEC (n = 3; B) ALI culture samples incubated with proinflammation mixtures demonstrated increased expression of Cp110 mRNA compared with that seen in sham-incubated cultures (TNF-α [10 ng/mL], KC or IL-8 [5 ng/mL], IL-6 [100 ng/mL], and IL-13 [100 ng/mL]). ★P ≤ .05. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Expression of Cp110 mRNA and protein in MSEC ALI cultures after incubation with individual cytokines. A, Incubation with murine TNF-α, KC, murine IL-6, and murine IL-13 all maintained increased expression levels of mCp110. B, Western blot analysis for mCp110 after cytokine exposure with semiquantitative analysis (lower panel), demonstrating significant upregulation of protein after cytokine exposure (TNF-α [10 ng/mL], KC or IL-8 [5 ng/mL], IL-6 [100 ng/mL], and IL-13 [100 ng/mL]). ★P ≤ .05, significantly different from the control 14th day. Journal of Allergy and Clinical Immunology  , e1DOI: ( /j.jaci ) Copyright © 2011 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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