Recently, much attention has been focused on ultra-structural abnormalities of cilia and their association with respiratory disease, infertility, and Kar-tageners syndrome (http://www.sciencedirect.com/science/article/pii/S0012369216382708). Although generally referred to as the immotile cilia syndrome, other observers, have identified motion in some of the cilia and suggested the use of the term “ciliary dysldnesis.” Sleigh has noted that at present, “primary ciliary dyskinesia” is the preferred term. This would encompass the previously described syndromes including Kartagener s syndrome and the immotile cilia syndrome. In addition to the originally noted lack of dynein arms, other more recently reported ultrastructural abnormalities have included defective radial spokes, ectopic position of microtubules, and abnormal numbers of microtubules with transposition of outer microtubules to the central area. Although first described in sperm, these ciliary abnormalities also affect mucus clearance from the bronchial tree, trachea and nose. In addition, these ciliary abnormalities are associated with disturbed function in the middle ear, sinuses, and meninges and situs inversus in half of afflicted human embryos.
In 1979, Afeelius noted that patients with the immotile cilia syndrome frequently developed nasal polyps and had spirometric readings which suggested obstructive airways disease. In addition, chronic sinus drainage was a prominent problem. Ibese findings suggested to us that asthmatics with nasal polyps and aspirin intolerance (“triad” asthma) might also have some type of ciliary abnormality, particularly since this seems to be a rather unique subpopulation of asthmatics who often have severe disease. If present, such an abnormality would be expected to aggravate the disturbed mucociliary clearance occurring at the beginning of asthmatic episodes. Accordingly, ultrastructural and functional studies were undertaken on the cilia of “triad” asthmatics and control subjects.
Methods
Nasal biopsies were obtained, with informed consent, (protocol was approved by the institutional Committee to Review Grants for Clinical Research and Investigation Involving Human Beings) from five male and two female adult nonsmokers with well documented histories of aspirin intolerance, asthma, and nasal polyps. In three cases, aspirin intolerance was confirmed by careful challenge in connection with another study. In addition, a semen sample was obtained from one patient, and as a positive control, a nasal biopsy specimen was obtained from an 18-year-old male subject with situs inversus and sinusitus. Two control specimens from adults undergoing nasal surgery for unrelated reasons also were included. Nasal punch biopsies were performed on normal-appearing areas of the middle and/or inferior turbinates after an application of 4 percent cocaine as a topical anesthetic. 
The biopsy specimens were immediately placed in Earle s balanced salt solution and then washed with phosphate buffered saline (PBS) containing 100 U/ml penicillin, 100 jig/ml streptomycin, and.02 fig/ml amphotericin B. After washing, the specimens were divided into approximately 2 mm x 2 mm pieces. Some pieces were placed into 2 percent gluteraldehyde and then processed for both scanning and transmission electron microscopy. The rest were examined by light microscopy for initial ciliary motility and then cultured at 37° С in Eagle s medium supplemented with 15 percent fetal calf serum, 2 mM glutamine, nonessential amino adds, 100 U/ml penicillin, 100 м-g/ml streptomycin, and 0.02 (ig/ml amphotericin B in a 5 percent COs atmosphere. Medium was changed weekly, and tests of ciliary activity were made periodically on bits of tissue removed from culture to hemocytometer chambers perfused with reagents dissolved in Earles balanced salt solution at 37° C. These bits of tissue were assessed for ciliary activity between one and nine days after being placed into tissue culture. An additional trial was performed with cilia removed 112 days after being placed into culture. Ciliary motility was scored visually on a scale of 0 to 3 in a manner similar to that employed by Forrest et al.
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