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Staphylococcus Enterotoxins In Cystic Fibrosis Patients With Chronic Rhinosinusitis
Abstracts S217
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Altered Airway Epithelial Cell Phenotype is Associated with Susceptibility to Allergic Asthma C. C. Lewis, K. Dienger, A. Sproles, M. Wills-Karp; Cincinnati Children’s Hospital Medical Center, Cincinnati, OH. RATIONALE: Asthma is a complex airway disease resulting from inappropriate Th2 inflammation. The airway epithelium (AEC) is first to encounter aeroallergens, exhibits activated phenotype in asthma, and has critical but undefined role(s) in the disease pathogenesis. We sought to determine the effects of allergen on AEC phenotype and to define the role(s) of AEC in mediating allergic responses. METHODS: We compared the AEC responses to house dust mite (HDM) in a well-characterized murine model of allergen-induced asthma, in which susceptible A/J mice develop the allergic phenotype, while resistant C3H/ HeJ do not. Primary tracheal AEC were isolated from A/J (n 5 8) and C3H (n 5 8), and cultured in presence or absence of HDM. Gene expression was determined via Affymetrix GeneChipÒ, and cytokine production measured via ELISA. RESULTS: 2707 genes were differentially expressed between strains in response to HDM. Compared to C3H, A/J AEC exhibited significant upregulation of genes involved in Th2 induction (IL23a, Tnc, Ii, Il1rl1), cytokines/chemokines (Csf2, Ccl20, Cx3cl1, Cxcl12), adhesion-mediated signaling (Itga2, Vcam1, Tspan18, Adam12), antigen presentation (H2Aa), and complement (C2, C1r). Conversely, C3H AEC had HDM-induced deficiencies in Il2rg, consistent with their diminished Th2 phenotype. ELISAs confirmed (p < 0.05) enhanced production of Csf2 and Ccl20, factors important in dendritic cell (DC) maturation and migration, respectively (Csf: AJ, 386.1 6 22.3; C3H, 90.6 6 9.5 pg/ml)(Ccl20: AJ: 640 6 6.6; C3H, 48 6 25 pg/ml). CONCLUSIONS: Our data suggest asthma may be associated with altered AEC phenotype, as AEC-derived factors may drive downstream immune responses with known relevance to the disease, including DC and Th2 cells.
834 835
[Withdrawn]
Chlamydophila Pneumoniae and Chronic Sinusitis J. Fahrenholz, C. Stratton, B. Wolf, J. Duncavage, Y. Tang, P. Russell, K. Fowler, D. Knighton, J. Cutler, M. Boruk; Vanderbilt University, Nashville, TN. RATIONALE: C. pneumoniae is an obligate intracellular organism that has been noted to cause persistent respiratory tract infections following an initial acute infection. Isolation of C. pneumoniae from the sinuses has been described in at least one case report. The specific aim of this investigation was to determine an association between C. pneumoniae infection and chronic sinusitis. METHODS: We offered participation to consecutive patients who consented for surgical therapy for medically refractory chronic sinusitis. We obtained sinus mucosal biopsy specimens from participants at the time of endoscopic sinus surgery. Patients undergoing septoplasty for deviated septum without associated sinus disease served as a control group. Qualitative PCR was performed to evaluate for the presence of C. pneumoniae in the biopsy samples. Quantitative real-time PCR was planned for any specimens with positive qualitative PCR results. RESULTS: Among 27 patients and 5 controls, we did not identify evidence of C. pneumoniae infection/colonization by qualitiative PCR analysis. CONCLUSIONS: Among patients with medically refractory chronic sinusitis who have failed antibiotic therapy, persistent C. pneumoniae infection does not appear to play a role. Funding: Vanderbilt University Medical Center
836
Staphylococcus Enterotoxins In Cystic Fibrosis Patients With Chronic Rhinosinusitis P. S. Hutcheson, J. F. Eisenbeis, D. S. Brink, E. M. Swierkosz, R. G. Slavin; Saint Louis University Medical School, Saint Louis, MO. RATIONALE: In patients with chronic rhinosinusitis with nasal polyps (CRSwNP) there is evidence that Staphylococcus aureus enterotoxins (SEs) contribute to the immune inflammatory response that characterizes the disease. CRSwNP occurs in cystic fibrosis (CF) patients who are frequently colonized with S. aureus, but the cellular infiltrate of the inflammatory response differs from CRSwNP non CF patients. We investigated the role of SEs in CF patients with CRSwNP and characterized the polypoid tissue inflammatory responses and each patient’s atopic status. METHODS: Blood was collected from 8 CF patients at the time of polypectomy. Antibody levels were determined by ImmunoCAP 100 and included IgE anti-SEA, -SEB, -SEC, -SED, -TSST-1, -Aspergillus, -Alternaria, -oak, -D. pteronyssinus, -ragweed, & -Kentucky blue grass, total IgE, and IgG anti-Alternaria. Culture and histopathology were performed on tissue specimens. RESULTS: Six of eight patients were colonized with S. aureus, but none had IgE against SEA, SEB, SEC, SED, or TSST-l. Inflammation was graded as moderate in 4 patients and marked in 4; lymphocytes and plasma cells represented the bulk of the inflammatory infiltrates with only rare or occasional neutrophils and eosinophils. Fungi were absent in both cultures and tissue sections. Total IgE ranged from 2-91 kU/L; 3 patients had IgE sensitivities: 1 to oak and Alternaria, 1 to Aspergillus and 1 to Alternaria. Seven of eight patients had low or undectable IgG antiAlternaria.. CONCLUSIONS: It appears that SEs do not elicit an IgE response in CF patients with CRSwNP, and thus do not contribute to the pathophysiology of the immune response. Funding: Saint Louis University
837
Distinct Immune Profiles Of Chronic Inflammatory, Eosinophilic, Aspirin Insensitive, And Fungal Sinusitis P. Huyett1, J. A. Negri1, J. Han2, S. Payne1, L. Borish1, J. W. Steinke1; 1 University of Virginia Health Systems, Charlottesville, VA, 2Eastern Virginia Medical School, Norfolk, VA. RATIONALE: Chronic sinusitis (CS) comprises several distinct disorders including those characterized by anatomical obstruction to the sinus ostia with chronic/recurrent infection and remodeling (chronic inflammatory sinusitis (CIS)), chronic hyperplastic sinusitis (CHS), aspirin-exacerbated respiratory disease (AERD), and allergic fungal sinusitis (AFS). Each can largely be distinguished clinically. We previously presented data regarding the histological characteristics of each condition. Our current studies address the unique immune fingerprints of these disorders. METHODS: CS tissue was obtained from the University of Virginia Department of Otolaryngology. RNA and protein were extracted from tissue and cytokine transcripts quantified via real time polymerase chain reaction (qPCR) or protein concentrations determined using ELISA. RESULTS: CIS is characterized by a chronic inflammatory infiltrate with dense collagen deposition. qPCR demonstrated markedly elevated expression of hypoxia-inducible factor 1a (HIF-1a) consistent with a role for hypoxemia in the pathophysiology of this condition. CIS tissue also ˆ , transcript levels. Supporting these findings, displayed increased TGF-bA ˆ ,1 protein compared to CHS or CIS had higher concentrations of TGF-bA AERD. As CHS comprises a Th2-associated cytokine disorder, increased expression of IL-4 and IL-13 were found in comparison to CIS. In contrast to CHS, AERD appears to be more of a mixed Th1/Th2 disorder with dramatically increased expression of IFN-g. Finally, consistent with the profuse allergic mucin deposition, AFS displayed the highest concentrations of Muc5. CONCLUSIONS: The various forms of CS can be identified by their transcript and protein profiles. These findings are consistent with the proposed pathophysiology of these disorders and suggest unique diseases with need for distinct therapeutic intervention. Funding: NIH, Richmond Eye and Ear Institute
MONDAY
833
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Altered Airway Epithelial Cell Phenotype is Associated with Susceptibility to Allergic Asthma C. C. Lewis, K. Dienger, A. Sproles, M. Wills-Karp; Cincinnati Children’s Hospital Medical Center, Cincinnati, OH. RATIONALE: Asthma is a complex airway disease resulting from inappropriate Th2 inflammation. The airway epithelium (AEC) is first to encounter aeroallergens, exhibits activated phenotype in asthma, and has critical but undefined role(s) in the disease pathogenesis. We sought to determine the effects of allergen on AEC phenotype and to define the role(s) of AEC in mediating allergic responses. METHODS: We compared the AEC responses to house dust mite (HDM) in a well-characterized murine model of allergen-induced asthma, in which susceptible A/J mice develop the allergic phenotype, while resistant C3H/ HeJ do not. Primary tracheal AEC were isolated from A/J (n 5 8) and C3H (n 5 8), and cultured in presence or absence of HDM. Gene expression was determined via Affymetrix GeneChipÒ, and cytokine production measured via ELISA. RESULTS: 2707 genes were differentially expressed between strains in response to HDM. Compared to C3H, A/J AEC exhibited significant upregulation of genes involved in Th2 induction (IL23a, Tnc, Ii, Il1rl1), cytokines/chemokines (Csf2, Ccl20, Cx3cl1, Cxcl12), adhesion-mediated signaling (Itga2, Vcam1, Tspan18, Adam12), antigen presentation (H2Aa), and complement (C2, C1r). Conversely, C3H AEC had HDM-induced deficiencies in Il2rg, consistent with their diminished Th2 phenotype. ELISAs confirmed (p < 0.05) enhanced production of Csf2 and Ccl20, factors important in dendritic cell (DC) maturation and migration, respectively (Csf: AJ, 386.1 6 22.3; C3H, 90.6 6 9.5 pg/ml)(Ccl20: AJ: 640 6 6.6; C3H, 48 6 25 pg/ml). CONCLUSIONS: Our data suggest asthma may be associated with altered AEC phenotype, as AEC-derived factors may drive downstream immune responses with known relevance to the disease, including DC and Th2 cells.
834 835
[Withdrawn]
Chlamydophila Pneumoniae and Chronic Sinusitis J. Fahrenholz, C. Stratton, B. Wolf, J. Duncavage, Y. Tang, P. Russell, K. Fowler, D. Knighton, J. Cutler, M. Boruk; Vanderbilt University, Nashville, TN. RATIONALE: C. pneumoniae is an obligate intracellular organism that has been noted to cause persistent respiratory tract infections following an initial acute infection. Isolation of C. pneumoniae from the sinuses has been described in at least one case report. The specific aim of this investigation was to determine an association between C. pneumoniae infection and chronic sinusitis. METHODS: We offered participation to consecutive patients who consented for surgical therapy for medically refractory chronic sinusitis. We obtained sinus mucosal biopsy specimens from participants at the time of endoscopic sinus surgery. Patients undergoing septoplasty for deviated septum without associated sinus disease served as a control group. Qualitative PCR was performed to evaluate for the presence of C. pneumoniae in the biopsy samples. Quantitative real-time PCR was planned for any specimens with positive qualitative PCR results. RESULTS: Among 27 patients and 5 controls, we did not identify evidence of C. pneumoniae infection/colonization by qualitiative PCR analysis. CONCLUSIONS: Among patients with medically refractory chronic sinusitis who have failed antibiotic therapy, persistent C. pneumoniae infection does not appear to play a role. Funding: Vanderbilt University Medical Center
836
Staphylococcus Enterotoxins In Cystic Fibrosis Patients With Chronic Rhinosinusitis P. S. Hutcheson, J. F. Eisenbeis, D. S. Brink, E. M. Swierkosz, R. G. Slavin; Saint Louis University Medical School, Saint Louis, MO. RATIONALE: In patients with chronic rhinosinusitis with nasal polyps (CRSwNP) there is evidence that Staphylococcus aureus enterotoxins (SEs) contribute to the immune inflammatory response that characterizes the disease. CRSwNP occurs in cystic fibrosis (CF) patients who are frequently colonized with S. aureus, but the cellular infiltrate of the inflammatory response differs from CRSwNP non CF patients. We investigated the role of SEs in CF patients with CRSwNP and characterized the polypoid tissue inflammatory responses and each patient’s atopic status. METHODS: Blood was collected from 8 CF patients at the time of polypectomy. Antibody levels were determined by ImmunoCAP 100 and included IgE anti-SEA, -SEB, -SEC, -SED, -TSST-1, -Aspergillus, -Alternaria, -oak, -D. pteronyssinus, -ragweed, & -Kentucky blue grass, total IgE, and IgG anti-Alternaria. Culture and histopathology were performed on tissue specimens. RESULTS: Six of eight patients were colonized with S. aureus, but none had IgE against SEA, SEB, SEC, SED, or TSST-l. Inflammation was graded as moderate in 4 patients and marked in 4; lymphocytes and plasma cells represented the bulk of the inflammatory infiltrates with only rare or occasional neutrophils and eosinophils. Fungi were absent in both cultures and tissue sections. Total IgE ranged from 2-91 kU/L; 3 patients had IgE sensitivities: 1 to oak and Alternaria, 1 to Aspergillus and 1 to Alternaria. Seven of eight patients had low or undectable IgG antiAlternaria.. CONCLUSIONS: It appears that SEs do not elicit an IgE response in CF patients with CRSwNP, and thus do not contribute to the pathophysiology of the immune response. Funding: Saint Louis University
837
Distinct Immune Profiles Of Chronic Inflammatory, Eosinophilic, Aspirin Insensitive, And Fungal Sinusitis P. Huyett1, J. A. Negri1, J. Han2, S. Payne1, L. Borish1, J. W. Steinke1; 1 University of Virginia Health Systems, Charlottesville, VA, 2Eastern Virginia Medical School, Norfolk, VA. RATIONALE: Chronic sinusitis (CS) comprises several distinct disorders including those characterized by anatomical obstruction to the sinus ostia with chronic/recurrent infection and remodeling (chronic inflammatory sinusitis (CIS)), chronic hyperplastic sinusitis (CHS), aspirin-exacerbated respiratory disease (AERD), and allergic fungal sinusitis (AFS). Each can largely be distinguished clinically. We previously presented data regarding the histological characteristics of each condition. Our current studies address the unique immune fingerprints of these disorders. METHODS: CS tissue was obtained from the University of Virginia Department of Otolaryngology. RNA and protein were extracted from tissue and cytokine transcripts quantified via real time polymerase chain reaction (qPCR) or protein concentrations determined using ELISA. RESULTS: CIS is characterized by a chronic inflammatory infiltrate with dense collagen deposition. qPCR demonstrated markedly elevated expression of hypoxia-inducible factor 1a (HIF-1a) consistent with a role for hypoxemia in the pathophysiology of this condition. CIS tissue also ˆ , transcript levels. Supporting these findings, displayed increased TGF-bA ˆ ,1 protein compared to CHS or CIS had higher concentrations of TGF-bA AERD. As CHS comprises a Th2-associated cytokine disorder, increased expression of IL-4 and IL-13 were found in comparison to CIS. In contrast to CHS, AERD appears to be more of a mixed Th1/Th2 disorder with dramatically increased expression of IFN-g. Finally, consistent with the profuse allergic mucin deposition, AFS displayed the highest concentrations of Muc5. CONCLUSIONS: The various forms of CS can be identified by their transcript and protein profiles. These findings are consistent with the proposed pathophysiology of these disorders and suggest unique diseases with need for distinct therapeutic intervention. Funding: NIH, Richmond Eye and Ear Institute
MONDAY
833