- Email: [email protected]
The German Mouse Clinic (gmc): A Systemic Phenotyping Platform To Uncover New Models For Allergic Diseases
Genetic Polymorphisms of Transforming Growth Factor-b Signaling Pathway and Kawasaki Disease in the Taiwanese population H. C. Kuo1, K. D. Yang1, W. C. Chang2; 1Kaohsiung Chang Gung Memorial Hospital, Taiwan, Kaohsiung, TAIWAN, 2Department of Medical Genetics, College of Medicine, Kaohsiung Medical University, Taiwan, Kaohsiung, TAIWAN. RATIONALE: Kawasaki disease is a systemic vasculitis associated with cardiovascular symptom. A previous study in the European descent has indicated that genetic variants of the transforming growth factor-b (TGF-b) signaling pathway are involved in the KD susceptibility and clinical status. This study was conducted to investigate if polymorphisms in transforming growth factor-b signaling pathway are associated with KD susceptibility, coronary artery lesion formation or intravenous immunoglobulin (IVIG) treatment responses. METHODS: A total of 907 subjects (381 KD patients and 526 controls) were investigated to identify 12 single nucleotide polymorphisms in transforming growth factor-b signaling pathway (TGFB2: rs2796817, rs10482751, rs2027567, rs12029576; TGFBR2: rs11466480; SMAD3: rs12901071, rs7162912, rs1438386, rs6494633, rs12910698, rs4776339) by using the TaqMan Allelic Discrimination assay. RESULTS: rs1438386, one of the 12 ingle nucleotide polymorphisms in the TGF-b signaling pathway is significantly associated with Kawasaki disease susceptibility. However, no significant associations between 12 ingle nucleotide polymorphisms and coronary artery lesion (CAL) formation or intravenous immunoglobulin (IVIG) treatment response were observed. Haplotype analysis did not yield any significant result. CONCLUSIONS: This study showed that genetic polymorphism rs1438386 associated with KD susceptibility, but not CAL formation, or IVIG treatment response in the Taiwanese population.
238
239
The German Mouse Clinic (gmc): A Systemic Phenotyping Platform To Uncover New Models For Allergic Diseases J. A. Aguilar-Pimentel1,2, T. Adler3, V. Gailus-Durner4, H. Fuchs4, J. Gutermuth2, C. B. Schmidt-Weber2, M. Hrab de Angelis4, D. H. Busch3, M. Ollert1,2; 1Technische Universitaet Muenchen, Department of Dermatology and Allergy Biederstein, GERMANY, 2Center of Allergy and Environment (ZAUM), Technische Universitaet Muenchen, GERMANY, 3 Technische Universitaet Muenchen, Institute for Medical Microbiology Immunology, GERMANY, 4Helmholtz Zentrum Muenchen, GmbH, Institute of Experimental Genetics, Munich, GERMANY. RATIONALE: A series of new candidate genes for allergic diseases has been uncovered by the use of a comprehensive systemic primary screen which covers, amongst others, the areas of immunology and allergy in the German Mouse Clinic (GMC). High-throughput strategies for the detection of allergy-prone mutant mice have been applied to identify phenotypic alterations in genetically modified mice with the aim to find new strategies for allergy diagnosis and anti-allergic therapy. METHODS: Total IgE levels in murine plasma are used as the first-line allergy-screening parameter. Mutant lines showing an interesting phenotype are subjected to a more in-depth assessment. This contains a wellestablished challenge-screen that includes a model of allergic sensitization and aerosol challenge. The subsequent analysis includes high-throughput quantification of immunoglobulins (bead-array technology), classification of cells from bronchoalveolar-lavage (flow-cytometry), immune phenotyping of lymphocytes, and quantification of multiple cytokines. RESULTS: This study has emerged new and interesting mouse mutant lines with particular allergy-phenotypes. Amongst the unexpected findings were sex-hormone-dependent levels of IgE, distinct T cell patterns after allergen-challenge in mice with a defect in intracellular vesicle transport, and a skewed airway inflammatory response in mutants with a defect in LPS-induced apoptosis caused by a gene of the SET domain family, known to be involved in methyl transferase activity. CONCLUSIONS: We have successfully revealed distinct gene functions in mutant mouse lines that enhance or reduce allergic disease in the murine model. These high-throughput technologies provide important advances
linking new and previously unexpected genes to the pathophysiology, diagnosis, and potential therapy of allergic disorders. (support: BMBFNGFNplus-01GS0868)
240
Exposure to Indoor Allergens in Urban Elementary Schools and Homes of Children with Asthma P. Permaul1, E. Hoffman2, C. Fu3, W. Sheehan4, S. Baxi4, J. Gaffin5, J. Lane6, A. Bailey3, D. Gold3, W. Phipatanakul4; 1Division of Pediatric Allergy and Immunology, Massachusetts General Hospital, Boston, MA, 2Department of Biostatistics, Harvard School of Public Health, Boston, MA, 3Channing Laboratory, Brigham and Women’s Hospital, Boston, MA, 4Division of Allergy and Immunology, Children’s Hospital Boston, Boston, MA, 5Division of Pulmonary, Children’s Hospital Boston, Boston, MA, 6Facilities Management, Boston, MA. RATIONALE: Most studies of indoor allergens have focused on home environment. However, schools may be an important site of allergen exposure for children with asthma. METHODS: Settled dust and airborne samples from classrooms, gymnasiums, and cafeterias in 12 inner-city schools were analyzed for indoor allergens by MARIA technology. School samples were linked to students with asthma enrolled in the School Inner-City Asthma Study to determine the presence of indoor allergens in their school environment. Bedroom settled dust samples of students were analyzed for indoor allergens in the same manner. RESULTS: From schools, 229 settled dust and 197 airborne samples were obtained. From homes, 118 settled dust samples were obtained. Generalized linear models using logit links showed significantly higher school settled dust levels of mouse (OR56.45, 95% CI: 3.60, 11.56, P < 0.0001), dog (OR52.44, 95% CI: 1.47, 4.04, P < 0.0008), and cat (OR52.98, 95% CI: 1.46, 6.09, P < 0.0032) as compared to homes. Less than 25% of enrolled students had pets at home. Settled dust mouse allergen levels in classrooms were moderately correlated with airborne levels (r 5 0.48; P < 0.0001). CONCLUSIONS: Inner-city children with asthma were exposed to higher levels of mouse, dog and cat allergen in their schools as compared to their homes, with Mus m 1 being the highest. There was a significant correlation between classroom settled dust and airborne Mus m 1 levels only. Further studies are needed to evaluate the role of school indoor allergen exposure and its effect on asthma morbidity in students with asthma.
241
Reduction in Dust Allergen Exposure through Healthy Homes Education F. Pacheco, M. G. Dhar, C. S. Barnes, C. Childrens Mercy Center for Environmental Health; Children’s Mercy Hospital, Kansas City, MO. RATIONALE: Asthma is typically related to exposure to protein allergen triggers found in house dust and indoor environments. To examine reductions in dust borne allergen triggers after institution of a Healthy Homes program the following studies were conducted. METHODS: Families with at least one asthmatic child were enrolled into a healthy homes program. Homes were evaluated for indoor and outdoor environmental conditions initially and subsequent to 6 months participation. Residents received education on maintaining a safe and healthy home and assistance in remediating unsafe or unhealthy conditions. Dust collections were taken in the asthmatic subject’s bedroom using a HUD developed protocol. Dust collections were analyzed for Fel d1, Can f1, Der f1, Der p1, Mus m1 and Bla g2 as well as fungal antigens from Alternaria, Aspergillus, Cladosporium and Penicillium. RESULTS: Initial mean dust measurements for allergens from the first 12 homes to fully complete the study varied from 7.6 mcg/g dust for Penicillium to 3.0 mcg/g dust for Aspergillus. Mean specific allergen values varied from 1.1 mcg/g for Mus m1 to 0.16 U/g for Bla g2. After at least 6 months participation dust borne allergen levels were reduced at least 50% for Fel d1, Penicillium, Bla g2 and Mus m1. Significant reductions were seen for Bla g2. Measured allergen levels remained unchanged or increased for Can f1, Der p1 and Aspergillus. CONCLUSIONS: Healthy homes education along with remediation can result in reduction in dust-borne allergen levels. More research is needed to understand specific factors related to the removal of dust borne allergenic material.
SATURDAY
Abstracts AB63
J ALLERGY CLIN IMMUNOL VOLUME 129, NUMBER 2
238
239
The German Mouse Clinic (gmc): A Systemic Phenotyping Platform To Uncover New Models For Allergic Diseases J. A. Aguilar-Pimentel1,2, T. Adler3, V. Gailus-Durner4, H. Fuchs4, J. Gutermuth2, C. B. Schmidt-Weber2, M. Hrab de Angelis4, D. H. Busch3, M. Ollert1,2; 1Technische Universitaet Muenchen, Department of Dermatology and Allergy Biederstein, GERMANY, 2Center of Allergy and Environment (ZAUM), Technische Universitaet Muenchen, GERMANY, 3 Technische Universitaet Muenchen, Institute for Medical Microbiology Immunology, GERMANY, 4Helmholtz Zentrum Muenchen, GmbH, Institute of Experimental Genetics, Munich, GERMANY. RATIONALE: A series of new candidate genes for allergic diseases has been uncovered by the use of a comprehensive systemic primary screen which covers, amongst others, the areas of immunology and allergy in the German Mouse Clinic (GMC). High-throughput strategies for the detection of allergy-prone mutant mice have been applied to identify phenotypic alterations in genetically modified mice with the aim to find new strategies for allergy diagnosis and anti-allergic therapy. METHODS: Total IgE levels in murine plasma are used as the first-line allergy-screening parameter. Mutant lines showing an interesting phenotype are subjected to a more in-depth assessment. This contains a wellestablished challenge-screen that includes a model of allergic sensitization and aerosol challenge. The subsequent analysis includes high-throughput quantification of immunoglobulins (bead-array technology), classification of cells from bronchoalveolar-lavage (flow-cytometry), immune phenotyping of lymphocytes, and quantification of multiple cytokines. RESULTS: This study has emerged new and interesting mouse mutant lines with particular allergy-phenotypes. Amongst the unexpected findings were sex-hormone-dependent levels of IgE, distinct T cell patterns after allergen-challenge in mice with a defect in intracellular vesicle transport, and a skewed airway inflammatory response in mutants with a defect in LPS-induced apoptosis caused by a gene of the SET domain family, known to be involved in methyl transferase activity. CONCLUSIONS: We have successfully revealed distinct gene functions in mutant mouse lines that enhance or reduce allergic disease in the murine model. These high-throughput technologies provide important advances
linking new and previously unexpected genes to the pathophysiology, diagnosis, and potential therapy of allergic disorders. (support: BMBFNGFNplus-01GS0868)
240
Exposure to Indoor Allergens in Urban Elementary Schools and Homes of Children with Asthma P. Permaul1, E. Hoffman2, C. Fu3, W. Sheehan4, S. Baxi4, J. Gaffin5, J. Lane6, A. Bailey3, D. Gold3, W. Phipatanakul4; 1Division of Pediatric Allergy and Immunology, Massachusetts General Hospital, Boston, MA, 2Department of Biostatistics, Harvard School of Public Health, Boston, MA, 3Channing Laboratory, Brigham and Women’s Hospital, Boston, MA, 4Division of Allergy and Immunology, Children’s Hospital Boston, Boston, MA, 5Division of Pulmonary, Children’s Hospital Boston, Boston, MA, 6Facilities Management, Boston, MA. RATIONALE: Most studies of indoor allergens have focused on home environment. However, schools may be an important site of allergen exposure for children with asthma. METHODS: Settled dust and airborne samples from classrooms, gymnasiums, and cafeterias in 12 inner-city schools were analyzed for indoor allergens by MARIA technology. School samples were linked to students with asthma enrolled in the School Inner-City Asthma Study to determine the presence of indoor allergens in their school environment. Bedroom settled dust samples of students were analyzed for indoor allergens in the same manner. RESULTS: From schools, 229 settled dust and 197 airborne samples were obtained. From homes, 118 settled dust samples were obtained. Generalized linear models using logit links showed significantly higher school settled dust levels of mouse (OR56.45, 95% CI: 3.60, 11.56, P < 0.0001), dog (OR52.44, 95% CI: 1.47, 4.04, P < 0.0008), and cat (OR52.98, 95% CI: 1.46, 6.09, P < 0.0032) as compared to homes. Less than 25% of enrolled students had pets at home. Settled dust mouse allergen levels in classrooms were moderately correlated with airborne levels (r 5 0.48; P < 0.0001). CONCLUSIONS: Inner-city children with asthma were exposed to higher levels of mouse, dog and cat allergen in their schools as compared to their homes, with Mus m 1 being the highest. There was a significant correlation between classroom settled dust and airborne Mus m 1 levels only. Further studies are needed to evaluate the role of school indoor allergen exposure and its effect on asthma morbidity in students with asthma.
241
Reduction in Dust Allergen Exposure through Healthy Homes Education F. Pacheco, M. G. Dhar, C. S. Barnes, C. Childrens Mercy Center for Environmental Health; Children’s Mercy Hospital, Kansas City, MO. RATIONALE: Asthma is typically related to exposure to protein allergen triggers found in house dust and indoor environments. To examine reductions in dust borne allergen triggers after institution of a Healthy Homes program the following studies were conducted. METHODS: Families with at least one asthmatic child were enrolled into a healthy homes program. Homes were evaluated for indoor and outdoor environmental conditions initially and subsequent to 6 months participation. Residents received education on maintaining a safe and healthy home and assistance in remediating unsafe or unhealthy conditions. Dust collections were taken in the asthmatic subject’s bedroom using a HUD developed protocol. Dust collections were analyzed for Fel d1, Can f1, Der f1, Der p1, Mus m1 and Bla g2 as well as fungal antigens from Alternaria, Aspergillus, Cladosporium and Penicillium. RESULTS: Initial mean dust measurements for allergens from the first 12 homes to fully complete the study varied from 7.6 mcg/g dust for Penicillium to 3.0 mcg/g dust for Aspergillus. Mean specific allergen values varied from 1.1 mcg/g for Mus m1 to 0.16 U/g for Bla g2. After at least 6 months participation dust borne allergen levels were reduced at least 50% for Fel d1, Penicillium, Bla g2 and Mus m1. Significant reductions were seen for Bla g2. Measured allergen levels remained unchanged or increased for Can f1, Der p1 and Aspergillus. CONCLUSIONS: Healthy homes education along with remediation can result in reduction in dust-borne allergen levels. More research is needed to understand specific factors related to the removal of dust borne allergenic material.
SATURDAY
Abstracts AB63
J ALLERGY CLIN IMMUNOL VOLUME 129, NUMBER 2