A mixed cell culture model for assessment of proliferation in tonsillar tissues from children with obstructive sleep apnea or recurrent tonsillitis.
BACKGROUND
Recurrent infective tonsillitis (RI) and obstructive sleep apnea (OSA) are the major indications for adenotonsillectomy (T&A) in children. However, little is known on the determinants of lymphadenoid tissue proliferation in the pediatric upper airway.


OBJECTIVE
To develop an in vitro culture system allowing for assessment of tonsillar or adenoidal proliferation under basal or stimulated conditions.


METHODS
Tonsils surgically removed from pediatric patients with obstructive sleep apnea and recurrent tonsillitis during T&A, were dissociated using standard methods. Whole cell tonsillar cultures were either maintained in normal medium or stimulated with lipopolysaccharide (25 microg/mL) and concanavalin A (10 microg/mL) for 24 hours (stimulated conditions [STIM]). Cellular proliferation was evaluated by [3H]thymidine incorporation. In parallel, supernatants were collected after 48 hours, and concentration of cytokines was measured using standard enzyme-linked immunosorbent assay procedures.


RESULTS
Basal proliferative rates were increased in the OSA group (305.2 +/- 40.6 cpm; n = 31) compared to RI group (232.8 +/- 31.9 cpm; n = 26; P < .001). No significant differences in proliferative rates emerged after STIM between OSA and RI. Furthermore, basal TNF-alpha, IL-6, and IL-8 concentrations in the supernatant were increased in OSA-derived cultures compared to RI, but IL-8 was higher after STIM in RI, while IL-6 remained increased in OSA.


CONCLUSIONS
The proliferative rates and concentrations of inflammatory mediators in tonsillar cell cultures from children with OSA and RI suggest that lymphadenoid tissue proliferation in these two conditions may be regulated by different mechanisms. This novel method may allow for future development of specific therapeutic interventions aimed at curtailing and reversing tonsillar and adenoidal hypertrophy in children in a disease-specific manner.
Serpero
Laura D
LD
Division of Sleep Medicine, Kosair Children's Hospital Research Institute, Department of Pediatrics, University of Louisville, Louisville, Kentucky, USA.
Kheirandish-Gozal
Leila
L
Dayyat
Ehab
E
Goldman
Julie L
JL
Kim
Jinkwan
J
Gozal
David
D
D000233:Adenoidectomy; D000234:Adenoids / Q000378:metabolism / Q000473:pathology*; D000704:Analysis of Variance; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002648:Child; D002675:Child, Preschool; D016207:Cytokines / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006984:Hypertrophy; D008297:Male; D014066:Palatine Tonsil / Q000378:metabolism / Q000473:pathology*; D012008:Recurrence; D020181:Sleep Apnea, Obstructive / Q000473:pathology / Q000601:surgery; D014068:Tonsillectomy; D014069:Tonsillitis / Q000473:pathology / Q000601:surgery
D016207:Cytokines