Treatment with rhDNase in patients with cystic fibrosis alters in-vitro CHIT-1 activity of isolated leucocytes.
Recent data suggest a possible relationship between cystic fibrosis (CF) pharmacotherapy, Aspergillus fumigatus colonization (AC) and/or allergic bronchopulmonary aspergillosis (ABPA). The aim of this study was to determine if anti-fungal defence mechanisms are influenced by CF pharmacotherapy, i.e. if (1) neutrophils form CF and non-CF donors differ in their ability to produce chitotriosidase (CHIT-1); (2) if incubation of isolated neutrophils with azithromycin, salbutamol, prednisolone or rhDNase might influence the CHIT-1 activity; and (3) if NETosis and neutrophil killing efficiency is influenced by rhDNase. Neutrophils were isolated from the blood of CF patients (n = 19; mean age 26·8 years or healthy, non-CF donors (n = 20; 38·7 years) and stimulated with phorbol-12-myristate-13-acetate (PMA), azithromycin, salbutamol, prednisolone or rhDNase. CHIT-1 enzyme activity was measured with a fluorescent substrate. NETosis was induced by PMA and neutrophil killing efficiency was assessed by a hyphae recovery assay. Neutrophil CHIT-1 activity was comparable in the presence or absence of PMA stimulation in both CF and non-CF donors. PMA stimulation and preincubation with rhDNase increased CHIT-1 activity in culture supernatants from non-CF and CF donors. However, this increase was significant in non-CF donors but not in CF patients (P < 0·05). RhDNase reduced the number of NETs in PMA-stimulated neutrophils and decreased the killing efficiency of leucocytes in our in-vitro model. Azithromycin, salbutamol or prednisolone had no effect on CHIT-1 activity. Stimulation of isolated leucocytes with PMA and treatment with rhDNase interfered with anti-fungal defence mechanisms. However, the impact of our findings for treatment in CF patients needs to be proved in a clinical cohort.
Department of Pediatric Allergy and Pulmonology, Clinic of Pediatrics UKSH, University of Luebeck, Luebeck.|Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL).
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Weckmann
Department of Pediatric Allergy and Pulmonology, Clinic of Pediatrics UKSH, University of Luebeck, Luebeck.|Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL).
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Schultheiss
Department of Pediatric Allergy and Pulmonology, Clinic of Pediatrics UKSH, University of Luebeck, Luebeck.|Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL).
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Hollaender
Clinic of Internal Medicine, UKSH, University of Kiel, Kiel.
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Bobis
Department of Molecular and Clinical Infectious Diseases, University of Luebeck, Luebeck, Germany.
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Rupp
Department of Pediatric Allergy and Pulmonology, Clinic of Pediatrics UKSH, University of Luebeck, Luebeck.|Airway Research Center North (ARCN), Member of the German Center for Lung Research (DZL).
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Kopp
D000293:Adolescent; D000328:Adult; D000368:Aged; D000420:Albuterol / Q000494:pharmacology / Q000627:therapeutic use; D001232:Aspergillus fumigatus / Q000302:isolation & purification; D017963:Azithromycin / Q000494:pharmacology / Q000627:therapeutic use; D001419:Bacteria / Q000302:isolation & purification; D003550:Cystic Fibrosis / Q000188:drug therapy / Q000276:immunology* / Q000382:microbiology; D003851:Deoxyribonucleases / Q000235:genetics / Q000627:therapeutic use*; D065206:Extracellular Traps / Q000187:drug effects; D005260:Female; D005658:Fungi / Q000302:isolation & purification; D006596:Hexosaminidases / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology* / Q000502:physiology; D010703:Phorbol Esters / Q000494:pharmacology; D011239:Prednisolone / Q000494:pharmacology / Q000627:therapeutic use; D013183:Sputum / Q000382:microbiology; D055815:Young Adult
D010703:Phorbol Esters; C119222:phorbol-12-myristate; D017963:Azithromycin; D011239:Prednisolone; D003851:Deoxyribonucleases; D006596:Hexosaminidases; C086400:chitotriosidase; D000420:Albuterol
fungal; human; inflammation; lung; neutrophils