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Il-4, Intron 3 Variable Number Tandem Repeats Analysis (vntr), And Il-13 -1055 Gene Polymorphism In Patients With Asthma And Copd In Venezuela
Abstracts S77
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Emergency Department Treated Asthmatics have Elevated Levels of Plasma Complement Split Products (CSP) and Peripheral Blood CSP Receptor (CSPr) Levels Greater than those from Outpatient Asthmatics H. Solomon, D. Dure, S. M. Chice, I. Kalampokis, M. Nowakowski, H. G. Durkin, R. Joks; SUNY Downstate Medical Center, Center for Allergy and Asthma Research, Brooklyn, NY. RATIONALE: Airway localized generation of the complement split products C5a and C3a may arise from inhalant allergens such as dust mite proteases (Maruo, 1997) and stimulants such as tobacco smoke (Kew, 1985) and may together potentiate airway responses leading to asthma exacerbation. This study determined whether plasma CSP and CSPr levels differ in adult asthmatics requiring ED treatment (ED1) from those measured from outpatient (OPD1) asthmatics and the effect of tobacco and allergy on these levels. METHODS: Plasma and peripheral blood cellular receptor levels of C5a desArg and C3a desArg (ELISA, flow cytometry) were obtained from ED1 (n 5 21)(during the ED visit) and OPD1 adult asthmatics (n 5 21). Current or previous tobacco smoking history (Tob1) and total serum IgE were determined. RESULTS: Levels of C3a des Arg, C5a desArg, and monocyte C3aR were significantly greater in the ED1 group vs. OPD1 (p 5 0.02, 0.04, and 0.05, respectively). Levels of lymphocytes CD88 (C5aR) approached significance in Tob1, regardless of treatment setting (p 5 0.08). Though plasma levels of both CSP were not elevated in Tob1, the Tob1, ED1 group had elevated levels of lymphocyte CD88, monocyte CD88, and lymphocyte C3aR compared to Tob-ED1 (p 5 0.03, 0.05, and 0.06, respectively). Total serum IgE did not differ among the groups. CONCLUSIONS: Levels of C5a desArg and C3a desArg are significantly elevated in ED1 asthmatics over those of OPD1 asthmatics. Tob1ED1 asthmatics have greater CSPr levels than Tob-ED1 asthmatics. This implies there is a degree of increased systemic inflammation in Tob1 ED1 asthmatics compared to Tob-, emergency department requiring asthmatics. Funding: New York State Research Grant
296
Il-4, Intron 3 Variable Number Tandem Repeats Analysis (vntr), And Il-13 -1055 Gene Polymorphism In Patients With Asthma And Copd In Venezuela N. E. Larocca1, D. Moreno2, F. Toro1, J. V. Garmendia1, J. B. De Sanctis1; 1 Instituto de Inmunologı´a. Universidad Central de Venezuela, Caracas, VENEZUELA, 2Catedra de Fisiopatologia. Escuela de Medicina Luis Razetti. Universidad Central de Venezuela, Caracas, VENEZUELA. RATIONALE: IL-4 and IL-13 cytokines involved in airway hyperresponsiveness, a critical event in patients with asthma and COPD. Studies in the variable number tandem repeats analysis (VNTR) in intron 3 of IL-4 and an SNP at the promoter region of IL13 (-1055) have not been documented in Venezuelan asthmatic and/or COPD patients. METHODS: Genomic DNA was purified from blood leukocytes by using a Quiagen kit and subsequently PCR was performed with specific primers to detect VNTR of 86 bp of IL-4 and RFLP to detect SNP in the IL-13 promoter region. The amplified products of PCR as well as the digested products were analyzed by polyacrylamide electrophoresis. The groups consisted in 100 patients with asthma, 100 with COPD and 100 age matched controls. RESULTS: Significant differences were encountered in the frequency of genotype RP2/RP2 for IL-4 between asthma and COPD (P 5 0.0304) and between asthma and controls (P 5 0.0137). In addition, TT genotype for IL-13 was more prevalent in asthmatic patients as compared to COPD patients (P 5 0.0013) and controls (P 5 0.04). CONCLUSIONS: The genotype RP2/RP2 for IL-4 and TT for IL-13 seem to be associated with susceptibility in patients with asthma in the Venezuelan population as compared with patients with COPD. Financed by FONACIT project # G2005000389. Funding: FONACIT
297
HLA Type and the Natural History of Asthma M. B. Martyn1, R. M. Jacobson2, G. A. Poland2, A. L. Weaver2, Y. J. Juhn2; 1Mayo Clinic Medical School, Rochester, MN, 2 Mayo Clinic, Rochester, MN. RATIONALE: This study investigated the influences of HLA genes on the natural history of asthma from the initial onset of symptoms to diagnosable asthma disease. METHODS: Study subjects were obtained from a sample of 340 healthy children, aged 5 to 12 years, who participated in a previous study and had HLA data available. Comprehensive medical record reviews were conducted to determine asthma status by applying predetermined criteria. We determined the duration from the initial onset of symptoms documented in medical records to the index date, when the subject first met the criteria for asthma disease. We examined 13 HLA genes previously reported to be associated with atopy or asthma, and then compared durations between carriers and non-carriers of HLA genes of interest. RESULTS: Among the 340 subjects with HLA data available, 117 children (34.4%) met the criteria for asthma. The median ages at onset of asthma symptoms and at the index date were 4.8 years and 7.3 years, respectively. The median durations between onset of symptom and index date for HLA DRB1*11 carriers and non-carriers were 552 vs. 61 days, respectively (p 5 0.004). Likewise, the same durations for HLA DQB1*0301 carriers and non-carriers were 420 vs. 60 days, respectively (p 5 0.013). On the other hand, HLA DQB1*0302 carriers had a shorter median duration, as compared to non-carriers (141 vs. 266 days, respectively, p 5 0.22). CONCLUSIONS: HLA type appears to influence the progression of asthma from initial symptoms to disease. Genetic factors may affect the natural history of asthma. Funding: Mayo Clinic
298
Antinuclear Antibodies in Asthma are Correlated with Lack of Asthma Control I. Agache1, L. Duca1, G. Pamfil1, M. Anghel2; 1Transylvania University Brasov, Faculty of Medicine, Brasov, ROMANIA, 2Brasov County Hospital, Immunology Dept., Brasov, ROMANIA. RATIONALE: Immune dysregulation in asthma and autoimmune diseases shares many common key pathogenic mechanisms: genetic background, mast cells and T cells, co-stimulatory molecules, cytokines and autoantibodies. Clinical significance of systemic autoimmunity in asthma is unclear. METHODS: 95 persistent asthma patients, mean age 53.52 6 13.7 years, 56% females, mean disease duration 13.21 6 12.30 years, were appreciated at inclusion for the presence of antinuclear antibodies - ANA (ELISA). ANA incidence in asthma was compared with 30 healthy subjects, matched for mean age and sex ratio. Patients were treated targeting asthma control and were strictly monitored for compliance to treatment. Asthma control (GINA 2006) was assessed after 1 year in relation with ANA, sputum and blood eosinophilia, NSAID intolerance, BMI > 25, chronic rhinosinusitis, smoking status, total serum IgE, asthma severity (ATS criteria) and psychopathology at inclusion and with compliance to treatment during the observation period. Statistics by multiple regression analysis and Chi-test. RESULTS: ANA incidence in asthma (20%) was significantly higher compared to the control group (3.3%) (p < 0.05). After 1 year 30(31.57%) patients achieved asthma control, 54(56.84%) were partly controlled and 11(11.57%) had uncontrolled asthma. Multiple regression analysis pointed out as independent risk patients factors for uncontrolled asthma: increased ANA titers (T-value 5 2.8206094; p 5 0.006), sputum eosinophilia (Tvalue 5 4.4305097; p < 0.001), blood eosinophilia (T-value 5 2.8999784, p 5 0.005), BMI > 25 (T-value 5 2.0803706; p 5 0.040), psychopathology (T-value 5 2.6056489; p 5 0.011) and non-compliance to treatment (T-value 5 2.1077154; p 5 0.038). CONCLUSIONS: The presence of antinuclear antibodies is an independent risk factor for uncontrolled asthma. A subset of asthma patients with difficult to control asthma can be identified by the presence of systemic autoimmunity.
SUNDAY
295
J ALLERGY CLIN IMMUNOL VOLUME 121, NUMBER 2
Emergency Department Treated Asthmatics have Elevated Levels of Plasma Complement Split Products (CSP) and Peripheral Blood CSP Receptor (CSPr) Levels Greater than those from Outpatient Asthmatics H. Solomon, D. Dure, S. M. Chice, I. Kalampokis, M. Nowakowski, H. G. Durkin, R. Joks; SUNY Downstate Medical Center, Center for Allergy and Asthma Research, Brooklyn, NY. RATIONALE: Airway localized generation of the complement split products C5a and C3a may arise from inhalant allergens such as dust mite proteases (Maruo, 1997) and stimulants such as tobacco smoke (Kew, 1985) and may together potentiate airway responses leading to asthma exacerbation. This study determined whether plasma CSP and CSPr levels differ in adult asthmatics requiring ED treatment (ED1) from those measured from outpatient (OPD1) asthmatics and the effect of tobacco and allergy on these levels. METHODS: Plasma and peripheral blood cellular receptor levels of C5a desArg and C3a desArg (ELISA, flow cytometry) were obtained from ED1 (n 5 21)(during the ED visit) and OPD1 adult asthmatics (n 5 21). Current or previous tobacco smoking history (Tob1) and total serum IgE were determined. RESULTS: Levels of C3a des Arg, C5a desArg, and monocyte C3aR were significantly greater in the ED1 group vs. OPD1 (p 5 0.02, 0.04, and 0.05, respectively). Levels of lymphocytes CD88 (C5aR) approached significance in Tob1, regardless of treatment setting (p 5 0.08). Though plasma levels of both CSP were not elevated in Tob1, the Tob1, ED1 group had elevated levels of lymphocyte CD88, monocyte CD88, and lymphocyte C3aR compared to Tob-ED1 (p 5 0.03, 0.05, and 0.06, respectively). Total serum IgE did not differ among the groups. CONCLUSIONS: Levels of C5a desArg and C3a desArg are significantly elevated in ED1 asthmatics over those of OPD1 asthmatics. Tob1ED1 asthmatics have greater CSPr levels than Tob-ED1 asthmatics. This implies there is a degree of increased systemic inflammation in Tob1 ED1 asthmatics compared to Tob-, emergency department requiring asthmatics. Funding: New York State Research Grant
296
Il-4, Intron 3 Variable Number Tandem Repeats Analysis (vntr), And Il-13 -1055 Gene Polymorphism In Patients With Asthma And Copd In Venezuela N. E. Larocca1, D. Moreno2, F. Toro1, J. V. Garmendia1, J. B. De Sanctis1; 1 Instituto de Inmunologı´a. Universidad Central de Venezuela, Caracas, VENEZUELA, 2Catedra de Fisiopatologia. Escuela de Medicina Luis Razetti. Universidad Central de Venezuela, Caracas, VENEZUELA. RATIONALE: IL-4 and IL-13 cytokines involved in airway hyperresponsiveness, a critical event in patients with asthma and COPD. Studies in the variable number tandem repeats analysis (VNTR) in intron 3 of IL-4 and an SNP at the promoter region of IL13 (-1055) have not been documented in Venezuelan asthmatic and/or COPD patients. METHODS: Genomic DNA was purified from blood leukocytes by using a Quiagen kit and subsequently PCR was performed with specific primers to detect VNTR of 86 bp of IL-4 and RFLP to detect SNP in the IL-13 promoter region. The amplified products of PCR as well as the digested products were analyzed by polyacrylamide electrophoresis. The groups consisted in 100 patients with asthma, 100 with COPD and 100 age matched controls. RESULTS: Significant differences were encountered in the frequency of genotype RP2/RP2 for IL-4 between asthma and COPD (P 5 0.0304) and between asthma and controls (P 5 0.0137). In addition, TT genotype for IL-13 was more prevalent in asthmatic patients as compared to COPD patients (P 5 0.0013) and controls (P 5 0.04). CONCLUSIONS: The genotype RP2/RP2 for IL-4 and TT for IL-13 seem to be associated with susceptibility in patients with asthma in the Venezuelan population as compared with patients with COPD. Financed by FONACIT project # G2005000389. Funding: FONACIT
297
HLA Type and the Natural History of Asthma M. B. Martyn1, R. M. Jacobson2, G. A. Poland2, A. L. Weaver2, Y. J. Juhn2; 1Mayo Clinic Medical School, Rochester, MN, 2 Mayo Clinic, Rochester, MN. RATIONALE: This study investigated the influences of HLA genes on the natural history of asthma from the initial onset of symptoms to diagnosable asthma disease. METHODS: Study subjects were obtained from a sample of 340 healthy children, aged 5 to 12 years, who participated in a previous study and had HLA data available. Comprehensive medical record reviews were conducted to determine asthma status by applying predetermined criteria. We determined the duration from the initial onset of symptoms documented in medical records to the index date, when the subject first met the criteria for asthma disease. We examined 13 HLA genes previously reported to be associated with atopy or asthma, and then compared durations between carriers and non-carriers of HLA genes of interest. RESULTS: Among the 340 subjects with HLA data available, 117 children (34.4%) met the criteria for asthma. The median ages at onset of asthma symptoms and at the index date were 4.8 years and 7.3 years, respectively. The median durations between onset of symptom and index date for HLA DRB1*11 carriers and non-carriers were 552 vs. 61 days, respectively (p 5 0.004). Likewise, the same durations for HLA DQB1*0301 carriers and non-carriers were 420 vs. 60 days, respectively (p 5 0.013). On the other hand, HLA DQB1*0302 carriers had a shorter median duration, as compared to non-carriers (141 vs. 266 days, respectively, p 5 0.22). CONCLUSIONS: HLA type appears to influence the progression of asthma from initial symptoms to disease. Genetic factors may affect the natural history of asthma. Funding: Mayo Clinic
298
Antinuclear Antibodies in Asthma are Correlated with Lack of Asthma Control I. Agache1, L. Duca1, G. Pamfil1, M. Anghel2; 1Transylvania University Brasov, Faculty of Medicine, Brasov, ROMANIA, 2Brasov County Hospital, Immunology Dept., Brasov, ROMANIA. RATIONALE: Immune dysregulation in asthma and autoimmune diseases shares many common key pathogenic mechanisms: genetic background, mast cells and T cells, co-stimulatory molecules, cytokines and autoantibodies. Clinical significance of systemic autoimmunity in asthma is unclear. METHODS: 95 persistent asthma patients, mean age 53.52 6 13.7 years, 56% females, mean disease duration 13.21 6 12.30 years, were appreciated at inclusion for the presence of antinuclear antibodies - ANA (ELISA). ANA incidence in asthma was compared with 30 healthy subjects, matched for mean age and sex ratio. Patients were treated targeting asthma control and were strictly monitored for compliance to treatment. Asthma control (GINA 2006) was assessed after 1 year in relation with ANA, sputum and blood eosinophilia, NSAID intolerance, BMI > 25, chronic rhinosinusitis, smoking status, total serum IgE, asthma severity (ATS criteria) and psychopathology at inclusion and with compliance to treatment during the observation period. Statistics by multiple regression analysis and Chi-test. RESULTS: ANA incidence in asthma (20%) was significantly higher compared to the control group (3.3%) (p < 0.05). After 1 year 30(31.57%) patients achieved asthma control, 54(56.84%) were partly controlled and 11(11.57%) had uncontrolled asthma. Multiple regression analysis pointed out as independent risk patients factors for uncontrolled asthma: increased ANA titers (T-value 5 2.8206094; p 5 0.006), sputum eosinophilia (Tvalue 5 4.4305097; p < 0.001), blood eosinophilia (T-value 5 2.8999784, p 5 0.005), BMI > 25 (T-value 5 2.0803706; p 5 0.040), psychopathology (T-value 5 2.6056489; p 5 0.011) and non-compliance to treatment (T-value 5 2.1077154; p 5 0.038). CONCLUSIONS: The presence of antinuclear antibodies is an independent risk factor for uncontrolled asthma. A subset of asthma patients with difficult to control asthma can be identified by the presence of systemic autoimmunity.
SUNDAY
295