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Tetsuji Takabayashi,
Atsushi Kato,
Anju T Peters,
Kathryn E Hulse,
Lydia A Suh,
Roderick Carter, James Norton,
Leslie C Grammer,
Bruce K Tan,
Rakesh K Chandra,
David B Conley,
Robert C Kern,
Shigeharu Fujieda,
Robert P Schleimer
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ABSTRACT: BACKGROUND: Profound edema or formation of a pseudocyst containing plasma proteins is a prominent characteristic of nasal polyps (NP). However, the mechanisms underlying NP retention of plasma proteins in the submucosa remain unclear. Recently, we reported that impairment of fibrinolysis causes excessive fibrin deposition in NP and this might be involved in the retention of plasma proteins. Although the coagulation cascade plays a critical role in fibrin clot formation at extravascular sites, the expression and role of coagulation factors in NP remain unclear. OBJECTIVE: The objective of this study was to investigate the expression of coagulation factors in patients with chronic rhinosinusitis (CRS). METHODS: Sinonasal tissues were collected from patients with CRS and control subjects. We assayed mRNA for factor XIII-A (FXIII-A) by using real-time PCR and measured FXIII-A protein by means of ELISA, immunohistochemistry, and immunofluorescence. RESULTS: FXIII-A mRNA levels were significantly increased in NP tissue from patients with CRS with NP (P < .001) compared with uncinate tissue from patients with CRS or control subjects. Similarly, FXIII-A protein levels were increased in NP. Immunofluorescence analysis revealed that FXIII-A expression in inflammatory cells and FXIII-A(+) cell numbers were significantly increased in NP. Most FXIII-A staining was observed within CD68(+)/CD163(+) M2 macrophages in NP. Levels of FXIII-A correlated with markers of M2 macrophages, suggesting that M2 macrophages are major FXIIIA-producing cells in NP. CONCLUSION: Overproduction of FXIII-A by M2 macrophages might contribute to the excessive fibrin deposition in the submucosa of NP, which might contribute to the tissue remodeling and pathogenesis of CRS with NP.
The Journal of allergy and clinical immunology 03/2013; · 9.17 Impact Factor
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Tetsuji Takabayashi,
Atsushi Kato,
Anju T Peters,
Kathryn E Hulse,
Lydia A Suh,
Roderick Carter, James Norton,
Leslie C Grammer,
Seong H Cho,
Bruce K Tan,
Rakesh K Chandra,
David B Conley,
Robert C Kern,
Shigeharu Fujieda,
Robert P Schleimer
[show abstract]
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ABSTRACT: RATIONALE: Nasal polyps (NP) are characterized by intense edema or formation of pseudocysts filled with plasma proteins, mainly albumin. However, the mechanisms underlying NP retention of plasma proteins in their submucosa remain unclear. OBJECTIVES: We hypothesized that formation of a fibrin mesh retains plasma proteins in NP. We assessed the fibrin deposition and expression of the components of the fibrinolytic system in patients with chronic rhinosinusitis (CRS). METHODS: We assessed fibrin deposition in nasal tissue from patients with CRS and control subjects by means of immunofluorescence. Fibrinolytic components, d-dimer and plasminogen activators were measured using ELISA, real-time PCR, and immunohistochemistry. We also performed gene expression and protein quantification analysis in cultured airway epithelial cells. MEASUREMENTS AND MAIN RESULTS: Immunofluorescence data showed profound fibrin deposition in NP compared with uncinate tissue (UT) from CRS and control subjects. Levels of the cross linked fibrin cleavage product protein, d-dimer, were significantly decreased in NP compared with UT from CRS and control, suggesting reduced fibrinolysis (P < .05). Expression levels of tissue plasminogen activator (t-PA) mRNA and protein were significantly decreased in NP compared with UT from CRS and control (P < .001). Immunohistochemistry demonstrated clear reduction of t-PA in NP, primarily in the epithelium and glands. Th2 cytokine stimulated cultured airway epithelial cells showed downregulation of t-PA, suggesting a potential Th2 mechanism in NP. CONCLUSIONS: A Th2-mediated reduction of t-PA might lead to excessive fibrin deposition in the submucosa of NP, which might contribute to the tissue remodeling and pathogenesis of CRS with nasal polyps (CRSwNP).
American Journal of Respiratory and Critical Care Medicine 11/2012; · 11.08 Impact Factor
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Tetsuji Takabayashi,
Atsushi Kato,
Anju T Peters,
Lydia A Suh,
Roderick Carter, James Norton,
Leslie C Grammer,
Bruce K Tan,
Rakesh K Chandra,
David B Conley,
Robert C Kern,
Shigeharu Fujieda,
Robert P Schleimer
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ABSTRACT: Although chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) is characterized by T(H)2 inflammation, the role of mast cells is poorly understood.
The objective of this study was to investigate the presence, localization, and phenotype of mast cells in patients with CRS.
We collected nasal tissue and nasal lavage fluid from patients with CRS and control subjects. We analyzed mRNA for the mast cell proteases tryptase, chymase, and carboxypeptidase A3 by using real-time PCR and measured mast cell protease proteins by using ELISA, immunohistochemistry, and immunofluorescence.
Tryptase mRNA was significantly increased in nasal polyps (NPs) from patients with CRSwNP (P< .001) compared with uncinate tissue from patients with CRS or control subjects. Tryptase protein was also elevated in NPs and in nasal lavage fluids from patients with CRSwNP. Immnohistochemistry showed increased numbers of mast cells in epithelium and glands but not within the lamina propria in NPs. The mast cells detected in the epithelium in NPs were characterized by the expression of tryptase and carboxypeptidase A3 but not chymase. Mast cells expressing all the 3 proteases were abundant within the glandular epithelium of NPs but were not found in normal glandular structures.
Herein we demonstrated a unique localization of mast cells within the glandular epithelium of NPs and showed that mast cells in NPs have distinct phenotypes that vary by tissue location. Glandular mast cells and the diverse subsets of mast cells detected may contribute to the pathogenesis of CRSwNP.
The Journal of allergy and clinical immunology 04/2012; 130(2):410-20.e5. · 9.17 Impact Factor
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Seong H Cho,
Seung J Hong,
Brian Han,
Sun H Lee,
Lydia Suh, James Norton,
David Lin,
David B Conley,
Rakesh Chandra,
Robert C Kern,
Bruce K Tan,
Atsushi Kato,
Anju Peters,
Leslie C Grammer,
Robert P Schleimer
The Journal of allergy and clinical immunology 03/2012; 129(3):858-860.e2. · 9.17 Impact Factor
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Bruce K Tan,
Quan-Zhen Li,
Lydia Suh,
Atsushi Kato,
David B Conley,
Rakesh K Chandra,
Jinchun Zhou, James Norton,
Roderick Carter,
Monique Hinchcliff,
Kathleen Harris,
Anju Peters,
Leslie C Grammer,
Robert C Kern,
Chandra Mohan,
Robert P Schleimer
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ABSTRACT: Chronic rhinosinusitis (CRS) with nasal polyps is an inflammatory condition of the nasal passage and paranasal sinuses characterized by T(H)2-biased inflammation with increased levels of B-cell activating factor of the TNF family (BAFF), B lymphocytes, and immunoglobulins. Because high levels of BAFF are associated with autoimmune diseases, we assessed for evidence of autoimmunity in patients with CRS.
The objective of this study was to investigate the presence of autoantibodies in sinonasal tissue from patients with CRS.
Standardized nasal tissue specimens were collected from patients with CRS and control subjects and assayed for immunoglobulin production, autoantibody levels, tissue distribution of immunoglobulins, and binding potential of antibodies in nasal tissue with a multiplexed autoantibody microarray, ELISA, and immunofluorescence.
Increased levels of several specific autoantibodies were found in nasal polyp tissue in comparison with levels seen in control tissue and inflamed tissue from patients with CRS without nasal polyps (P < .05). In particular, nuclear-targeted autoantibodies, such as anti-dsDNA IgG and IgA antibodies, were found at increased levels in nasal polyps (P < .05) and particularly in nasal polyps from patients requiring revision surgery for recurrence. Direct immunofluorescence staining demonstrated diffuse epithelial and subepithelial deposition of IgG and increased numbers of IgA-secreting plasma cells not seen in control nasal tissue.
Autoantibodies, particularly those against nuclear antigens, are present at locally increased levels in nasal polyps. The presence of autoantibodies suggests that the microenvironment of a nasal polyp promotes the expansion of self-reactive B-cell clones. Although the pathogenicity of these antibodies remains to be elucidated, the presence of increased anti-dsDNA antibody levels is associated with a clinically more aggressive form of CRS with nasal polyps requiring repeated surgery.
The Journal of allergy and clinical immunology 12/2011; 128(6):1198-1206.e1. · 9.17 Impact Factor
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Sarah Peterson,
Julie A Poposki,
Deepti R Nagarkar,
Regina T Chustz,
Anju T Peters,
Lydia A Suh,
Roderick Carter, James Norton,
Kathleen E Harris,
Leslie C Grammer,
Bruce K Tan,
Rakesh K Chandra,
David B Conley,
Robert C Kern,
Robert P Schleimer,
Atsushi Kato
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ABSTRACT: Chronic rhinosinusitis with nasal polyps (CRSwNP) is associated with T(H)2-dominant inflammation, including eosinophilia, which is in contrast to chronic rhinosinusitis (CRS) without nasal polyps (NPs). CC chemokine ligand 18 (CCL18)/pulmonary and activation-regulated chemokine is known to recruit naive T cells, B cells, and immature dendritic cells, as well as to activate fibroblasts. CCL18 is thought to be involved in T(H)2-related inflammatory diseases, including asthma and atopic dermatitis.
The objective of this study was to investigate the expression of CCL18 in patients with CRS.
Using NP tissue and uncinate tissue (UT) from control subjects and patients with CRS, we examined the expression of CCL18 mRNA using real-time PCR and measured CCL18 protein using ELISA, Western blotting, and immunofluorescence.
Compared with UT tissue from control subjects, CCL18 mRNA levels were significantly increased in NPs (P < .001) and UT (P < .05) from patients with CRSwNP but not in UT from patients with CRS without NPs. Similarly, CCL18 protein levels were increased in NPs and UT from patients with CRSwNP, and levels were even higher in patients with Samter's triad. Immunohistochemical analysis revealed CCL18 expression in inflammatory cells, and CCL18(+) cell numbers were significantly increased in NPs. Immunofluorescence data showed colocalization of CCL18 in CD68(+)/CD163(+)/macrophage mannose receptor-positive M2 macrophages and tryptase-positive mast cells in NPs. Levels of CCL18 correlated with markers of M2 macrophages but not with tryptase levels, suggesting that M2 macrophages are major CCL18-producing cells in NPs.
Overproduction of CCL18 might contribute to the pathogenesis of CRSwNP through its known activities, which include recruitment of lymphocytes and dendritic cells, activation of fibroblasts, and initiation of local inflammation.
The Journal of allergy and clinical immunology 09/2011; 129(1):119-27.e1-9. · 9.17 Impact Factor
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Julie A Poposki,
Ashraf Uzzaman,
Deepti R Nagarkar,
Regina T Chustz,
Anju T Peters,
Lydia A Suh,
Roderick Carter, James Norton,
Kathleen E Harris,
Leslie C Grammer,
Bruce K Tan,
Rakesh K Chandra,
David B Conley,
Robert C Kern,
Robert P Schleimer,
Atsushi Kato
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ABSTRACT: Chronic rhinosinusitis (CRS) is a heterogeneous chronic disease characterized by local inflammation of the sinonasal tissues. The pathogenesis of CRS remains controversial, but it has been associated with the accumulation of various immune and inflammatory cells in sinus tissue.
The objective of this study was to investigate the expression of the chemokine CCL23, which is known to bind to CCR1 and recruit monocytes, macrophages, and dendritic cells, in patients with CRS.
We collected nasal tissue from patients with CRS and control subjects. We assayed mRNA for CCL23 by using real-time PCR and measured CCL23 protein by means of ELISA, immunohistochemistry, and immunofluorescence.
CCL23 mRNA levels were significantly increased in nasal polyps (NPs) from patients with CRS with nasal polyps (CRSwNP; P < .05) compared with inferior turbinate and uncinate tissue from patients with CRS or control subjects. CCL23 protein levels were also increased in NPs, although these levels were not statistically significant. Immunohistochemical analysis revealed CCL23 expression in mucosal epithelial cells and inflammatory cells, but accumulation of CCL23(+) inflammatory cells occurred only in NPs. Immunofluorescence data showed CCL23 colocalization with eosinophil cationic protein-positive eosinophils. The concentration of CCL23 in NPs positively correlated with the concentration of eosinophil cationic protein, suggesting that eosinophils are major CCL23-producing cells in NPs. Finally, we found that CCL23 protein levels were significantly increased in NPs from patients with CRSwNP with aspirin sensitivity.
Overproduction of CCL23 in NPs might contribute to the pathogenesis of eosinophilic CRSwNP through the recruitment of CCR1(+) inflammatory cells, including monocytes and macrophages, and the amplification of local inflammation.
The Journal of allergy and clinical immunology 04/2011; 128(1):73-81.e4. · 9.17 Impact Factor
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David D Tieu,
Anju T Peters,
Roderick G Carter,
Roderick T Carter,
Lydia Suh,
David B Conley,
Rakesh Chandra, James Norton,
Leslie C Grammer,
Kathleen E Harris,
Atsushi Kato,
Robert C Kern,
Robert P Schleimer
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ABSTRACT: Decreased epithelial expression of mRNA for S100A7 (psoriasin) and S100A8/A9 (calprotectin) has been reported in patients with chronic rhinosinusitis (CRS).
We sought to assess whether the expression of S100 proteins is also altered in the sinonasal cavity of patients with CRS.
We determined levels of S100 proteins in nasal lavage fluid and sinonasal tissue extracts from patients with CRS using ELISA and immunohistochemical analysis of nasal polyp tissue from patients with CRS with nasal polyps and uncinate tissue from healthy control subjects, patients with CRSsNP, and patients with CRSwNP.
Expression levels of S100 proteins were decreased compared with those seen in control subjects in nasal lavage fluid from both CRS groups (P < .05). Similarly, tissue expression of these proteins assessed by means of immunohistochemistry demonstrated clear reductions, primarily in the epithelial lining. Interestingly, levels of calprotectin were increased in nasal polyp tissue despite lower levels in lavage fluid. Levels of calprotectin in nasal tissues were correlated with levels of neutrophils, as assessed by means of quantification of neutrophil elastase.
Several S100 proteins are in the epidermal differentiation complex of genes and have been demonstrated to play a role in maintenance of barrier function and formation of an antimicrobial shield. We demonstrate significantly decreased levels of expression of S100 proteins in the epithelium of patients with CRS, which might lead to diminished innate immune responses and barrier function. Increased levels of calprotectin in nasal polyp tissue might reflect neutrophil recruitment and a compensatory mechanism. Future studies will be important to determine whether reduced levels of S100 proteins lead to decreased antimicrobial responses in the upper airways and sinuses and whether this reduction plays a causative role in CRS pathogenesis and susceptibility to infectious disease.
The Journal of allergy and clinical immunology 03/2010; 125(3):667-75. · 9.17 Impact Factor
