Chitotriosidase is the primary active chitinase in the human lung and is modulated by genotype and smoking habit Max A. Seibold, PhD, Samantha Donnelly,

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Chitotriosidase is the primary active chitinase in the human lung and is modulated by genotype and smoking habit Max A. Seibold, PhD, Samantha Donnelly, PhD, Margaret Solon, BA, Anh Innes, MD, Prescott G. Woodruff, MD, MPH, Rolf G. Boot, PhD, Esteban González Burchard, MD, MPH, John V. Fahy, MD, MSc Journal of Allergy and Clinical Immunology Volume 122, Issue 5, Pages 944-950.e3 (November 2008) DOI: 10.1016/j.jaci.2008.08.023 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Effect of pH and disease status on chitinase activity in 77 BAL samples. A, Points represent chitinase activity measured for each BAL sample by using the synthetic chitin substrate 4-MU-(4-deoxy)chitobiose in McIlvaine's buffer at pH 2.2, 4.6, and 7.0. The line represents the median value. B, Points represent chitinase activity measured for each BAL sample (at pH 4.6) stratified by disease status. The line represents the median value. ∗Significantly greater than the healthy group, P < .0001. Journal of Allergy and Clinical Immunology 2008 122, 944-950.e3DOI: (10.1016/j.jaci.2008.08.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Effect of CHIT1 24-bp duplication genotype status on chitinase activity in BAL fluid. Points represent chitinase activity measured at pH 4.6 for subjects with a no-duplication CHIT1 genotype and for subjects heterozygous and homozygous for the CHIT1 duplication. Chitinase activity was undetectable in subjects homozygous for the duplication and reduced in subjects heterozygous for the duplication. Journal of Allergy and Clinical Immunology 2008 122, 944-950.e3DOI: (10.1016/j.jaci.2008.08.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 AMCase gene expression in lavage fluid macrophages from healthy subjects, asthmatic subjects, and habitual smokers. A, AMCase protein domain structure, with region spliced out in TSA1902S transcript denoted by arrows and dotted lines. Red arrows denote region amplified by RT-PCR primer set 1, and blue arrows denote region amplified by RT-PCR primer set 2. B, AMCase gene expression stratified by red primer set 1 (active site–containing transcripts) and blue primer set 2 (all AMCase transcripts). Journal of Allergy and Clinical Immunology 2008 122, 944-950.e3DOI: (10.1016/j.jaci.2008.08.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 AMCase protein in BAL fluid from healthy subjects (n = 13), asthmatic subjects (n = 16), and habitual smokers (n = 19). The y-axis shows values generated from densitometric analysis of protein bands relative to the intensity of control AMCase protein. The line represents the median value. ∗Significantly less than the healthy group, P = .023. Journal of Allergy and Clinical Immunology 2008 122, 944-950.e3DOI: (10.1016/j.jaci.2008.08.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 CHIT1 gene expression and correlation with BAL fluid chitinase activity. A, CHIT1 gene expression in bronchial brushings stratified by disease status. ∗Significantly greater than the healthy group, P < .05. B, CHIT1 gene expression in lavage fluid macrophages stratified by disease status. ∗Significantly greater than the healthy group, P < .05. C, Scatterplot of CHIT1 macrophage gene expression by BAL fluid chitinase activity at pH 4.6. Journal of Allergy and Clinical Immunology 2008 122, 944-950.e3DOI: (10.1016/j.jaci.2008.08.023) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions