Abstract
Primary ciliary dyskinesia (PCD) is a heterogeneous genetic disorder characterized by recurrent airway infections and situs inversus in half of affected individuals. Diagnosis currently relies on demonstration of abnormal ciliary ultrastructure or altered ciliary beat. Alterations encountered in secondary ciliary dyskinesia (SCD) caused by inflammation often complicate the diagnostic workup. We have recently shown that in respiratory epithelial cells from PCD patients with outer dynein arm defects the dynein protein DNAH5 is mislocalized and either completely or partially absent from the ciliary axoneme. In this study, we addressed the question whether SCD might affect axonemal DNAH5 localization in respiratory cells. To induce SCD in vitro, we treated primary human respiratory epithelial cell cultures with interleukin-13 (IL-13). Ciliary function and ultrastructure were assessed by high-speed videomicroscopy and transmission electron microscopy, respectively. For in vivo localization of DNAH5, we performed nasal brushing biopsies in patients with evidence of SCD. Expression of DNAH5 was analyzed by immunofluorescence microscopy. IL-13–treated cells showed evidence of SCD. Ciliary beat frequency was significantly reduced and ultrastructural analyses showed axonemal disorganization compared with control cells. High-resolution immunofluorescence studies of respiratory epithelial cells with SCD identified in vitro and in vivo normal axonemal DNAH5 localization. DNAH5 localization is not altered by SCD, indicating a high potential for immunofluorescence analysis as a novel diagnostic tool in PCD.
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Abbreviations
- IL-13:
-
interleukin-13
- PCD:
-
primary ciliary dyskinesia
- SCD:
-
secondary ciliary dyskinesia
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Acknowledgements
The authors thank R. Nitschke and S. Haxelmans, Life Imaging Center, Institute for Biology I, University Freiburg, for their excellent support with confocal microscopy. They are grateful to T. Willems and M. Jorissen, Otorhinolaryngology, Department for Human Genetics, Leuven, Belgium for help in establishing the monolayer and suspension cell culture technique of the respiratory epithelium cells in our laboratory. The authors thank J. Kalnitski and M. Krome for excellent technical assistance and R. Laszig, Department of Otorhinolaryngology, University of Freiburg, Freiburg, Germany, for supporting this study.
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This work was supported by grant P-LS-AL/7 from the Landesstiftung Baden-Württemberg.
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Olbrich, H., Horváth, J., Fekete, A. et al. Axonemal Localization of the Dynein Component DNAH5 Is Not Altered in Secondary Ciliary Dyskinesia. Pediatr Res 59, 418–422 (2006). https://doi.org/10.1203/01.pdr.0000200809.21364.e2
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DOI: https://doi.org/10.1203/01.pdr.0000200809.21364.e2
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