- Email: [email protected]
Development of an animal model for allergic conjunctivitis: Influence of genetic factors and allergen concentration on the immune response
Budesonide Nasal Washings Improve Outcome in Refractory Chronic Hyperplastic Sinusitis D. Cheung, J. King, I. Hussain; Allergy and Immunology, Washington University School of Medicine, Saint Louis, MO. RATIONALE: Chronic hyperplastic sinusitis with nasal polyposis (CHS/NP) is commonly refractory to treatments such as intranasal steroid sprays or saline nasal washes and many patients require oral steroids. We examined the use of budesonide nasal washings for CHS/NP refractory to traditional therapy. METHODS: Patients with CHS/NP, average Lund-Mackay score of 15, at a tertiary care referral center were started on budesonide nasal washings in 2003. We retrospectively examined the frequency of systemic antibiotic and steroid use, symptom score, and rhinoscopy findings before and after this intervention. Paired t-test was used for statistical analysis. RESULTS: Eight patients (M=6, W=2, mean age=48, all refractory to sinus surgeries and antimicrobial nasal washings) were included. The median time on budesonide was 12 months. All patients had symptom score reduction during treatment (19.0±5.7 vs. 14.1±6.6, n=8, p=0.007). The number of antibiotic courses prescribed on budesonide was also reduced compared to the 6 months prior to treatment (2.3±2.0 vs. 0.9±1.1, n=8, p=0.008). 7 patients were on oral steroids prior to treatment. 4 were off steroids and 3 had significant oral steroid dose reduction on treatment. The one patient not on oral steroids prior to budesonide had 3 courses of oral steroids in the 6 months prior to intervention, but only 1 course after. Rhinoscopy during treatment showed decreased edema, decreased mucopurulent secretions, and shrinkage of polypoid tissue. CONCLUSION: In patients with CHS/NP refractory to standard therapy, budesonide nasal wash may decrease symptoms and antibiotic use. Budesonide nasal washings also allowed weaning of oral steroids and may decrease polyp recurrence.
514
Development of an Animal Model for Allergic Conjunctivitis: Influence of Genetic Factors and Allergen Concentration on the Immune Response P. Giavina-Bianchi, J. Kalil, L. V. Rizzo; Clinical Immunology and Allergy, FMUSP, São Paulo, BRAZIL. RATIONALE: Animal models of diseases are extremely important in studying the physiopathogenesis of human pathologies and for testing novel therapeutic interventions. The aims of the present study are: (i) to develop an animal model that simulates human allergic conjunctivitis; and (ii) to study how the allergic response may be influenced by the allergen dose used for immunization and by genetic factors. METHODS: Sixty mice of the C57Bl/6 strain and sixty of the BALB/c strain were immunized, receiving 5 g, 50 g, or 500 g of allergen. After ocular challenge, mice were examined in order to clinically verify the occurrence or not of conjunctivitis. The material obtained from the animals was used for total and specific IgE and IgG1 dosage, for assays of Dpt-specific lymphocyte proliferation and supernatant cytokine dosage, and for histopathological evaluation of the conjunctiva. RESULTS: We have developed a murine model of allergic conjunctivitis induced by the Dermatophagoides pteronyssinus. The model is similar to human disease clinically and from laboratory findings. In the mouse, conjunctivitis was associated with a Th2 cytokine profile. However, IL-10 seemed involved with disease blockade in association with IFN. CONCLUSIONS: The murine model developed is suitable for the study of immunopathogenesis and as a template for future therapies. Using BALB/c and C57BL/6 mice, we demonstrated that genetic factors play a role in both determining susceptibility and resistance but also to establish the allergen concentration needed to induce or to block disease development. Funding: FMUSP
515
Effects of Antihistamines on Human Erythrocyte Plasma Membrane Integrity and Phosphatidylserine Distribution In Vitro G. Graff1, J. M. Yanni1, M. M. Momsen2, P. C. Schmid2, H. L. Brockman2; 1Ophthalmic Program Research, Alcon Research Ltd., Fort Worth, TX, 2Hormel Institute, University of Minnesota, Austin, MN.
516
RATIONALE: To explore loss of phosphatidylserine (PS) membrane asymmetry as part of the lytic mechanism of Ketotifen, Azelastine and Epinastine membrane interactions compared to the non-lytic Olopatadine. METHODS: Changes in PS plasma membrane asymmetry were determined by FACS analysis using fluorescein isothiocyanate (FITC) tagged annexin V. Enzymatic relocation of PS from the outer to the inner membrane leaflet was prevented by N-ethylmaleimide (NEM). Membrane leakage was quantified by hemoglobin release or by the appearance of a FTIC-annexin V labeled cell population RESULTS: Exposure of NEM-treated erythrocytes to Olopatadine (0.110 mM) did not affect cell particle size, membrane permeability, or PS membrane localization. Ca2+- ion and ionophore (A23187) addition to NEM-treated erythrocytes (positive control) yielded a small population of permeabilized cells with changed particle size, in addition to an intact cell population that exhibited FITC-annexin V binding due to PS translocation to the outer membrane. Exposure of NEM-treated cells to Ketotifen, and Azelastine revealed erythrocyte permeabilization without translocation of PS to the outer membrane leaflet prior to cell lysis. Epinastine induced a significant transfer of PS from the inner to the outer membrane leaflet at concentrations below those causing cell permeabilization. CONCLUSIONS: Ketotifen and Azelastine cause erythrocyte membrane permeabilization without a prior disturbance of membrane PS asymmetry. Epinastine induces a significant disturbance in PS membrane asymmetry at sub-lytic concentrations. Oloptadine is totally devoid of membrane perturbing effects. Disturbance of membrane phospholipid asymmetry may be relevant to topical ocular use since PS exposure on the outer membrane leaflet has been reported to target cells for apoptosis. Funding: Alcon Research Ltd Impact on the Quality of Life of the Addition of Topical Ocular Anti-Allergic Therapy to Existing Systemic and/or Nasal Therapy in a Population of Allergic Rhinitis Patients Using RQLQ and ACQLQ Questionnaires W. Storms1, M. Beck2, S. Kimura3, T. Westbrook4, W. Berger5, S. Galant6; 1PC and Research Center, Colorado Springs, CO, 2Asthma Allergy Associates, Miami, FL, 3Cordova Allergy, Pensacola, FL, 4Allergy and Asthma Center of NW Florida, Pensacola, FL, 5Southern California Research, Mission Viejo, CA, 6Clinical Trials of Orange County, Inc., Orange, CA. RATIONALE: This study used two questionnaires to examine quality of life (QoL) during allergy season in allergic rhinitis patients on concomitant systemic and/or nasal therapy. The impact of the addition of a topical ocular anti-allergic (olopatadine HCl 0.1% ophthalmic solution) to concomitant anti-allergic therapy was evaluated. METHODS: This was a 4-week, multi-center, prospective, open-label, cross-over QoL environmental study. Diagnosed rhinitis patients currently on systemic and/or topical nasal therapy and with no prior diagnosis of allergic conjunctivitis or usage of prescription ocular allergic therapy were enrolled. Patients attended three office visits and were asked to complete two QoL questionnaires (RQLQ and ACQLQ) at each visit. Patients continued their prescribed rhinitis treatment regimen throughout the trial. RESULTS: 200 patients completed this study. At baseline these patients had global scores of RQLQ: 2.16 and ACQLQ: 2.15. The number of days during the previous week that patients experienced eye allergy symptoms was 3.92. Following the addition of ocular treatment, clinically relevant and statistically significant improvement was seen in nasal and ocular global scores (RQLQ: -1.00, ACQLQ:-1.19(p<0.01)). Clinical significance was defined as 0.5 score improvement on a 6.0 scale. On average, patients experienced 2.2 fewer days with ocular allergic symptoms. These results were consistent across concomitant rhinitis treatment groups. CONCLUSIONS: QoL, as measured by both nasal and ocular domains, was improved following addition of topical ocular anti-allergic therapy to concomitant rhinitis therapy. These data suggest that opportunity exists to improve QoL in patients without prior allergic conjunctivitis diagnosis or use of prescription ocular allergic therapy. Funding: Alcon Research, Ltd.
517
SUNDAY
Abstracts S129
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2
514
Development of an Animal Model for Allergic Conjunctivitis: Influence of Genetic Factors and Allergen Concentration on the Immune Response P. Giavina-Bianchi, J. Kalil, L. V. Rizzo; Clinical Immunology and Allergy, FMUSP, São Paulo, BRAZIL. RATIONALE: Animal models of diseases are extremely important in studying the physiopathogenesis of human pathologies and for testing novel therapeutic interventions. The aims of the present study are: (i) to develop an animal model that simulates human allergic conjunctivitis; and (ii) to study how the allergic response may be influenced by the allergen dose used for immunization and by genetic factors. METHODS: Sixty mice of the C57Bl/6 strain and sixty of the BALB/c strain were immunized, receiving 5 g, 50 g, or 500 g of allergen. After ocular challenge, mice were examined in order to clinically verify the occurrence or not of conjunctivitis. The material obtained from the animals was used for total and specific IgE and IgG1 dosage, for assays of Dpt-specific lymphocyte proliferation and supernatant cytokine dosage, and for histopathological evaluation of the conjunctiva. RESULTS: We have developed a murine model of allergic conjunctivitis induced by the Dermatophagoides pteronyssinus. The model is similar to human disease clinically and from laboratory findings. In the mouse, conjunctivitis was associated with a Th2 cytokine profile. However, IL-10 seemed involved with disease blockade in association with IFN. CONCLUSIONS: The murine model developed is suitable for the study of immunopathogenesis and as a template for future therapies. Using BALB/c and C57BL/6 mice, we demonstrated that genetic factors play a role in both determining susceptibility and resistance but also to establish the allergen concentration needed to induce or to block disease development. Funding: FMUSP
515
Effects of Antihistamines on Human Erythrocyte Plasma Membrane Integrity and Phosphatidylserine Distribution In Vitro G. Graff1, J. M. Yanni1, M. M. Momsen2, P. C. Schmid2, H. L. Brockman2; 1Ophthalmic Program Research, Alcon Research Ltd., Fort Worth, TX, 2Hormel Institute, University of Minnesota, Austin, MN.
516
RATIONALE: To explore loss of phosphatidylserine (PS) membrane asymmetry as part of the lytic mechanism of Ketotifen, Azelastine and Epinastine membrane interactions compared to the non-lytic Olopatadine. METHODS: Changes in PS plasma membrane asymmetry were determined by FACS analysis using fluorescein isothiocyanate (FITC) tagged annexin V. Enzymatic relocation of PS from the outer to the inner membrane leaflet was prevented by N-ethylmaleimide (NEM). Membrane leakage was quantified by hemoglobin release or by the appearance of a FTIC-annexin V labeled cell population RESULTS: Exposure of NEM-treated erythrocytes to Olopatadine (0.110 mM) did not affect cell particle size, membrane permeability, or PS membrane localization. Ca2+- ion and ionophore (A23187) addition to NEM-treated erythrocytes (positive control) yielded a small population of permeabilized cells with changed particle size, in addition to an intact cell population that exhibited FITC-annexin V binding due to PS translocation to the outer membrane. Exposure of NEM-treated cells to Ketotifen, and Azelastine revealed erythrocyte permeabilization without translocation of PS to the outer membrane leaflet prior to cell lysis. Epinastine induced a significant transfer of PS from the inner to the outer membrane leaflet at concentrations below those causing cell permeabilization. CONCLUSIONS: Ketotifen and Azelastine cause erythrocyte membrane permeabilization without a prior disturbance of membrane PS asymmetry. Epinastine induces a significant disturbance in PS membrane asymmetry at sub-lytic concentrations. Oloptadine is totally devoid of membrane perturbing effects. Disturbance of membrane phospholipid asymmetry may be relevant to topical ocular use since PS exposure on the outer membrane leaflet has been reported to target cells for apoptosis. Funding: Alcon Research Ltd Impact on the Quality of Life of the Addition of Topical Ocular Anti-Allergic Therapy to Existing Systemic and/or Nasal Therapy in a Population of Allergic Rhinitis Patients Using RQLQ and ACQLQ Questionnaires W. Storms1, M. Beck2, S. Kimura3, T. Westbrook4, W. Berger5, S. Galant6; 1PC and Research Center, Colorado Springs, CO, 2Asthma Allergy Associates, Miami, FL, 3Cordova Allergy, Pensacola, FL, 4Allergy and Asthma Center of NW Florida, Pensacola, FL, 5Southern California Research, Mission Viejo, CA, 6Clinical Trials of Orange County, Inc., Orange, CA. RATIONALE: This study used two questionnaires to examine quality of life (QoL) during allergy season in allergic rhinitis patients on concomitant systemic and/or nasal therapy. The impact of the addition of a topical ocular anti-allergic (olopatadine HCl 0.1% ophthalmic solution) to concomitant anti-allergic therapy was evaluated. METHODS: This was a 4-week, multi-center, prospective, open-label, cross-over QoL environmental study. Diagnosed rhinitis patients currently on systemic and/or topical nasal therapy and with no prior diagnosis of allergic conjunctivitis or usage of prescription ocular allergic therapy were enrolled. Patients attended three office visits and were asked to complete two QoL questionnaires (RQLQ and ACQLQ) at each visit. Patients continued their prescribed rhinitis treatment regimen throughout the trial. RESULTS: 200 patients completed this study. At baseline these patients had global scores of RQLQ: 2.16 and ACQLQ: 2.15. The number of days during the previous week that patients experienced eye allergy symptoms was 3.92. Following the addition of ocular treatment, clinically relevant and statistically significant improvement was seen in nasal and ocular global scores (RQLQ: -1.00, ACQLQ:-1.19(p<0.01)). Clinical significance was defined as 0.5 score improvement on a 6.0 scale. On average, patients experienced 2.2 fewer days with ocular allergic symptoms. These results were consistent across concomitant rhinitis treatment groups. CONCLUSIONS: QoL, as measured by both nasal and ocular domains, was improved following addition of topical ocular anti-allergic therapy to concomitant rhinitis therapy. These data suggest that opportunity exists to improve QoL in patients without prior allergic conjunctivitis diagnosis or use of prescription ocular allergic therapy. Funding: Alcon Research, Ltd.
517
SUNDAY
Abstracts S129
J ALLERGY CLIN IMMUNOL VOLUME 115, NUMBER 2