- Email: [email protected]
Lack of Association Between the Tumor Necrosis Factor-α Regulatory Region Genetic Polymorphisms Associated With Elevated Tumor Necrosis Factor-α Levels and Children With Asthma
tissue fibrosis by selectively stimulating and activating transforming growth factor 1. J Exp Med 2001; 194:809 – 821
Identification of Genes and Proteins Regulated by Interleukin-5 in Human Eosinophils Using Microarrays and Two-Dimensional Electrophoresis/Mass Spectrometry* Sara Håkansson, BS; Karin Behrens, BS; Gyo¨ rgy Marko-Varga, PhD; Henrik Lindberg, BS; Stefan Pierrou, PhD; and Witte Koopmann, PhD
Lack of Association Between the Tumor Necrosis Factor-␣ Regulatory Region Genetic Polymorphisms Associated With Elevated Tumor Necrosis Factor-␣ Levels and Children With Asthma* Lama El Bahlawan, MD; Michael Christensen, PhD; Said Binaei, BS; Cathy Murphy, BS; Qing Zhang, BS; and Micheal Quasney, MD, PhD
(CHEST 2003; 123:374S) Abbreviation: IL ⫽ interleukin
(CHEST 2003; 123:374S–375S) Abbreviation: TNF ⫽ tumor necrosis factor
cellular events underlying the development and T hemaintenance of allergic airway disease are complex.
A number of different inflammatory cell types, including lymphocytes, mast cells, and eosinophils, are thought to play key roles in allergic airway disease. The accumulation of eosinophils in the airway is one of the hallmarks of asthma, and a large body of work has focused on understanding the biology of eosinophils in patients with asthma. Interleukin (IL)-5 is a crucial factor in the biology of eosinophils and has been shown to regulate a number of eosinophil functions, including apoptosis, chemotaxis, and exocytosis. In order to identify genes and proteins that are regulated by IL-5, we have performed proteomic and genomic analyses of human eosinophils that had and had not been treated with IL-5. In one set of experiments, eosinophils were prepared from the peripheral blood of five healthy donors and were stimulated in vitro with IL-5 or vehicle. Protein extracts were resolved on two-dimensional gels, and differentially expressed proteins were excised, trypsinized, and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. In parallel experiments, microarrays (U95A; Affymetrix; Santa Clara, CA) were run on pooled RNA from primary human eosinophils that had and had not been treated with IL-5. Genes expressed in nonstimulated eosinophils were identified, as were subsets of genes that had been either up-regulated or down-
*From the Department of Molecular Sciences, AstraZeneca R&D Lund, Lund, Sweden. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Witte Koopmann, PhD, the Department of Molecular Sciences, AstraZeneca R&D Lund, S-221 87 Lund, Sweden 374S
regulated by IL-5 treatment. The results from the genomics analysis were compared with the results from the proteomics analysis.
polymorphisms influence the magnitude of the G enetic cytokine response after the occurrence of various
inflammatory stimuli. The pathophysiology of asthma involves, in part, the inflammation of the airway epithelium. Tumor necrosis factor (TNF)-␣, a proinflammatory cytokine, is released into the airway and serum of patients with asthma. Indeed, higher serum levels of TNF-␣ have been demonstrated in children with active disease. We tested the hypothesis that children with asthma have genetic polymorphisms in the regulatory region of the gene coding for TNF-␣ that are associated with higher levels of TNF-␣.
Materials and Methods Blood samples were obtained from children with exacerbations of asthma. Genotypes were determined by polymerase chain reaction amplification and restriction enzyme digestion. Genotypic frequencies were determined and compared with samples from healthy control subjects (Table 1). Statistical analysis was performed using 2 analysis.
Results Thirty-eight children with asthma were enrolled into the study. The genotypic frequencies of the polymorphic *From the Departments of Pediatrics (Drs. Bahlawan and Quasney, and Mr. Zhang) and Clinical Pharmacy (Drs. Christensen and Mr. Binaei); Children’s Foundation Research Center, University of Tennessee, Memphis TN; and LeBonheur Children’s Medical Center (Ms. Murphy), Memphis, TN. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Micheal Quasney, MD, PhD, Department of Pediatrics, 50 N Dunlap St, Memphis TN 38103; e-mail: [email protected]
Thomas L. Petty 45th Annual Aspen Lung Conference: Asthma in the New Millennium
Table 1—Genotypic Frequencies at Three Polymorphic Sites* TNF-␣ ⫺238
TNF-␣ ⫺308
LT-␣ ⫹250
Group
G/G
G/A
A/A
G/G
G/A
A/A
G/G
G/A
A/A
Control subjects (n ⫽ 231) Asthma patients (n ⫽ 38)
0.917 0.868
0.083 0.105
0 0.026
0.788 0.816
0.195 0.132
0.017 0.053
0.216 0.158
0.476 0.553
0.307 0.289
*LT ⫽ lymphotoxin.
lymphotoxin-␣⫹250, TNF-␣-238, and TNF-␣-308 sites were determined. There was no significant difference between the genotypic frequencies of the polymorphisms that were associated with the higher secretion of TNF-␣ and children with asthma.
Conclusions In summary, children with asthma do not have an increase in the frequencies of the various polymorphisms associated with higher levels of TNF-␣. Further study is warranted because of the small number of children with asthma who have been evaluated thus far and the wide variety of polymorphisms that may play a role.
reaction amplification and restriction enzyme digestion. Genotypic frequencies were determined and compared with samples from healthy control subjects. Statistical analysis was performed using 2 analysis.
Results Thirty-eight children with status asthmaticus were enrolled into the study. The frequencies of the polymorphic sites at amino acid positions 16 and 27 in the 2-adrenergic receptor are shown below. The frequency of the Gly/Gly polymorphism at position 16 was significantly lower in children with status asthmaticus when compared with that of a healthy control group (0.184 vs 0.348, respectively; p ⬍ 0.05) [Table 1].
Conclusions
2ⴚAdrenergic Receptor Polymorphisms in Children With Status Asthmaticus* Saeed Binaei, BS; Michael Christensen, PhD; Cathy Murphy, RN; Qing Zhang, BS; and Michael Quasney, MD, PhD
(CHEST 2003; 123:375S) polymorphisms within the gene coding for the G enetic  -adrenergic receptor have been identified that in2
fluence cell signaling and down-regulation. The Gly/Gly polymorphism at position 16 is associated with increased airway hyperactivity, while the Glu/Glu polymorphism at position 27 is associated with decreased airway hyperactivity. We tested the hypothesis that children with status asthmaticus have genetic polymorphisms that are associated with increased airway hyperactivity.
Materials and Methods Blood samples were obtained from children with exacerbations of asthma. Genotypes were determined by polymerase chain *From the Departments of Clinical Pharmacy (Mr. Binaei and Dr. Christensen) and Pediatrics (Mr. Zhang and Dr. Quasney), Children’s Foundation Research Center, University of Tennessee, Memphis, TN; LeBonheur Children’s Medical Center (Ms. Murphy), Memphis, TN. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Michael Quasney, MD, PhD, College of Medicine, Department of Pediatrics, 50 North Dunlap, Memphis, TN 38103; e-mail: [email protected] www.chestjournal.org
Children with status asthmaticus have a lower frequency of the Gly/Gly polymorphism at amino acid position 16 in the 2-adrenergic receptor. Further study is warranted because of the small number of children with asthma who have been evaluated thus far and the wide variety of polymorphisms that may play a role.
Table 1—Frequencies of Polymorphism in the 2-Adrenergic Receptor in Children With Asthma and Control Subjects AA Position 16 Variables
AA Position 27
Arg/Arg Arg/Gly Gly/Gly Gln/Gln Gln/Glu Glu/Glu
Control subjects 0.219 (n ⫽ 155) Asthma patients 0.184 (n ⫽ 38)
0.432
0.348
0.692
0.231
0.077
0.632
0.184
0.605
0.342
0.053
Interleukin-13 Induces Surfactant Function Abnormality in the Murine Lung* Zhou Zhu, MD, PhD; Goran Enhorning, MD, PhD; Tao Zheng, MD; Qingsheng Chen, MD; Ning Yuan Chen, BS; Robert Homer, MD, PhD; and Jack A. Elias, MD
(CHEST 2003; 123:375S–376S) Abbreviations: IL ⫽ interleukin
CLSE ⫽ calf
lung
surfactant
CHEST / 123 / 3 / MARCH, 2003 SUPPLEMENT
extract;
375S
Identification of Genes and Proteins Regulated by Interleukin-5 in Human Eosinophils Using Microarrays and Two-Dimensional Electrophoresis/Mass Spectrometry* Sara Håkansson, BS; Karin Behrens, BS; Gyo¨ rgy Marko-Varga, PhD; Henrik Lindberg, BS; Stefan Pierrou, PhD; and Witte Koopmann, PhD
Lack of Association Between the Tumor Necrosis Factor-␣ Regulatory Region Genetic Polymorphisms Associated With Elevated Tumor Necrosis Factor-␣ Levels and Children With Asthma* Lama El Bahlawan, MD; Michael Christensen, PhD; Said Binaei, BS; Cathy Murphy, BS; Qing Zhang, BS; and Micheal Quasney, MD, PhD
(CHEST 2003; 123:374S) Abbreviation: IL ⫽ interleukin
(CHEST 2003; 123:374S–375S) Abbreviation: TNF ⫽ tumor necrosis factor
cellular events underlying the development and T hemaintenance of allergic airway disease are complex.
A number of different inflammatory cell types, including lymphocytes, mast cells, and eosinophils, are thought to play key roles in allergic airway disease. The accumulation of eosinophils in the airway is one of the hallmarks of asthma, and a large body of work has focused on understanding the biology of eosinophils in patients with asthma. Interleukin (IL)-5 is a crucial factor in the biology of eosinophils and has been shown to regulate a number of eosinophil functions, including apoptosis, chemotaxis, and exocytosis. In order to identify genes and proteins that are regulated by IL-5, we have performed proteomic and genomic analyses of human eosinophils that had and had not been treated with IL-5. In one set of experiments, eosinophils were prepared from the peripheral blood of five healthy donors and were stimulated in vitro with IL-5 or vehicle. Protein extracts were resolved on two-dimensional gels, and differentially expressed proteins were excised, trypsinized, and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. In parallel experiments, microarrays (U95A; Affymetrix; Santa Clara, CA) were run on pooled RNA from primary human eosinophils that had and had not been treated with IL-5. Genes expressed in nonstimulated eosinophils were identified, as were subsets of genes that had been either up-regulated or down-
*From the Department of Molecular Sciences, AstraZeneca R&D Lund, Lund, Sweden. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Witte Koopmann, PhD, the Department of Molecular Sciences, AstraZeneca R&D Lund, S-221 87 Lund, Sweden 374S
regulated by IL-5 treatment. The results from the genomics analysis were compared with the results from the proteomics analysis.
polymorphisms influence the magnitude of the G enetic cytokine response after the occurrence of various
inflammatory stimuli. The pathophysiology of asthma involves, in part, the inflammation of the airway epithelium. Tumor necrosis factor (TNF)-␣, a proinflammatory cytokine, is released into the airway and serum of patients with asthma. Indeed, higher serum levels of TNF-␣ have been demonstrated in children with active disease. We tested the hypothesis that children with asthma have genetic polymorphisms in the regulatory region of the gene coding for TNF-␣ that are associated with higher levels of TNF-␣.
Materials and Methods Blood samples were obtained from children with exacerbations of asthma. Genotypes were determined by polymerase chain reaction amplification and restriction enzyme digestion. Genotypic frequencies were determined and compared with samples from healthy control subjects (Table 1). Statistical analysis was performed using 2 analysis.
Results Thirty-eight children with asthma were enrolled into the study. The genotypic frequencies of the polymorphic *From the Departments of Pediatrics (Drs. Bahlawan and Quasney, and Mr. Zhang) and Clinical Pharmacy (Drs. Christensen and Mr. Binaei); Children’s Foundation Research Center, University of Tennessee, Memphis TN; and LeBonheur Children’s Medical Center (Ms. Murphy), Memphis, TN. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Micheal Quasney, MD, PhD, Department of Pediatrics, 50 N Dunlap St, Memphis TN 38103; e-mail: [email protected]
Thomas L. Petty 45th Annual Aspen Lung Conference: Asthma in the New Millennium
Table 1—Genotypic Frequencies at Three Polymorphic Sites* TNF-␣ ⫺238
TNF-␣ ⫺308
LT-␣ ⫹250
Group
G/G
G/A
A/A
G/G
G/A
A/A
G/G
G/A
A/A
Control subjects (n ⫽ 231) Asthma patients (n ⫽ 38)
0.917 0.868
0.083 0.105
0 0.026
0.788 0.816
0.195 0.132
0.017 0.053
0.216 0.158
0.476 0.553
0.307 0.289
*LT ⫽ lymphotoxin.
lymphotoxin-␣⫹250, TNF-␣-238, and TNF-␣-308 sites were determined. There was no significant difference between the genotypic frequencies of the polymorphisms that were associated with the higher secretion of TNF-␣ and children with asthma.
Conclusions In summary, children with asthma do not have an increase in the frequencies of the various polymorphisms associated with higher levels of TNF-␣. Further study is warranted because of the small number of children with asthma who have been evaluated thus far and the wide variety of polymorphisms that may play a role.
reaction amplification and restriction enzyme digestion. Genotypic frequencies were determined and compared with samples from healthy control subjects. Statistical analysis was performed using 2 analysis.
Results Thirty-eight children with status asthmaticus were enrolled into the study. The frequencies of the polymorphic sites at amino acid positions 16 and 27 in the 2-adrenergic receptor are shown below. The frequency of the Gly/Gly polymorphism at position 16 was significantly lower in children with status asthmaticus when compared with that of a healthy control group (0.184 vs 0.348, respectively; p ⬍ 0.05) [Table 1].
Conclusions
2ⴚAdrenergic Receptor Polymorphisms in Children With Status Asthmaticus* Saeed Binaei, BS; Michael Christensen, PhD; Cathy Murphy, RN; Qing Zhang, BS; and Michael Quasney, MD, PhD
(CHEST 2003; 123:375S) polymorphisms within the gene coding for the G enetic  -adrenergic receptor have been identified that in2
fluence cell signaling and down-regulation. The Gly/Gly polymorphism at position 16 is associated with increased airway hyperactivity, while the Glu/Glu polymorphism at position 27 is associated with decreased airway hyperactivity. We tested the hypothesis that children with status asthmaticus have genetic polymorphisms that are associated with increased airway hyperactivity.
Materials and Methods Blood samples were obtained from children with exacerbations of asthma. Genotypes were determined by polymerase chain *From the Departments of Clinical Pharmacy (Mr. Binaei and Dr. Christensen) and Pediatrics (Mr. Zhang and Dr. Quasney), Children’s Foundation Research Center, University of Tennessee, Memphis, TN; LeBonheur Children’s Medical Center (Ms. Murphy), Memphis, TN. Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (e-mail: [email protected]). Correspondence to: Michael Quasney, MD, PhD, College of Medicine, Department of Pediatrics, 50 North Dunlap, Memphis, TN 38103; e-mail: [email protected] www.chestjournal.org
Children with status asthmaticus have a lower frequency of the Gly/Gly polymorphism at amino acid position 16 in the 2-adrenergic receptor. Further study is warranted because of the small number of children with asthma who have been evaluated thus far and the wide variety of polymorphisms that may play a role.
Table 1—Frequencies of Polymorphism in the 2-Adrenergic Receptor in Children With Asthma and Control Subjects AA Position 16 Variables
AA Position 27
Arg/Arg Arg/Gly Gly/Gly Gln/Gln Gln/Glu Glu/Glu
Control subjects 0.219 (n ⫽ 155) Asthma patients 0.184 (n ⫽ 38)
0.432
0.348
0.692
0.231
0.077
0.632
0.184
0.605
0.342
0.053
Interleukin-13 Induces Surfactant Function Abnormality in the Murine Lung* Zhou Zhu, MD, PhD; Goran Enhorning, MD, PhD; Tao Zheng, MD; Qingsheng Chen, MD; Ning Yuan Chen, BS; Robert Homer, MD, PhD; and Jack A. Elias, MD
(CHEST 2003; 123:375S–376S) Abbreviations: IL ⫽ interleukin
CLSE ⫽ calf
lung
surfactant
CHEST / 123 / 3 / MARCH, 2003 SUPPLEMENT
extract;
375S