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. Objectives: To develop an in vit...

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Veröffentlicht in:The Laryngoscope 2009-05, Vol.119 (5), p.1005-1010
Hauptverfasser: Serpero, Laura D., Kheirandish-Gozal, Leila, Dayyat, Ehab, Goldman, Julie L., Kim, Jinkwan, Gozal, David
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container_end_page 1010
container_issue 5
container_start_page 1005
container_title The Laryngoscope
container_volume 119
creator Serpero, Laura D.
Kheirandish-Gozal, Leila
Dayyat, Ehab
Goldman, Julie L.
Kim, Jinkwan
Gozal, David
description 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. Objectives: 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 μg/mL) and concanavalin A (10 μg/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. Laryngoscope, 2009
doi_str_mv 10.1002/lary.20147
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However, little is known on the determinants of lymphadenoid tissue proliferation in the pediatric upper airway. Objectives: 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&amp;A, were dissociated using standard methods. Whole cell tonsillar cultures were either maintained in normal medium or stimulated with lipopolysaccharide (25 μg/mL) and concanavalin A (10 μg/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 &lt; .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. 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However, little is known on the determinants of lymphadenoid tissue proliferation in the pediatric upper airway. Objectives: 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&amp;A, were dissociated using standard methods. Whole cell tonsillar cultures were either maintained in normal medium or stimulated with lipopolysaccharide (25 μg/mL) and concanavalin A (10 μg/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 &lt; .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. 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Stomatology</subject><subject>Palatine Tonsil - metabolism</subject><subject>Palatine Tonsil - pathology</subject><subject>Pneumology</subject><subject>Recurrence</subject><subject>recurrent tonsillitis</subject><subject>Respiratory system : syndromes and miscellaneous diseases</subject><subject>Sleep Apnea, Obstructive - pathology</subject><subject>Sleep Apnea, Obstructive - surgery</subject><subject>tonsillar hypertrophy</subject><subject>Tonsillectomy</subject><subject>Tonsillitis - pathology</subject><subject>Tonsillitis - surgery</subject><issn>0023-852X</issn><issn>1531-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcuOEzEQRS0EYkJgwwcgb2CB1IPdtvuxQYpGENBEIPEQj41VcZeJwd0Odvc8voMfxiEhwIZVLerUrVt1CbnP2SlnrHziIV6flozL-gaZcSV4IdtW3SSz3BRFo8qPJ-ROSl8Z47VQ7DY54W1ZVaqRM_JjQXt3hR016D01kx-niLQPHXpqQ6SQEqbU4zDSYOk2Bu8sRhhdGKgb6BiG5Hw2QEeX0oSJ2hh6ajbOdxEHeunGDQ3rNMbJjO4CafKIWwrbAYFm-YhminGnflByWecuuWXBJ7x3qHPy_vmzd2cvitXr5cuzxaowsqnqwoKQwG2NUorKtqqqAG1XIwgDNeddB0ytuTAC2pI1knetZUZxBIlViQ2KOXm6191O6x47k21E8HobXZ8_qgM4_W9ncBv9JVzosmlLWfMs8OggEMP3fPyoe5d2j4QBw5R0VfOayRzCnDzegyaGlCLa4xLO9C5DvctQ_8owww_-tvUHPYSWgYcHAJIBbyMMxqUjV3KpRMN39vieu3Qer_-zUq8Wbz79Xl7sZ1wa8eo4A_FbvkbUSn94tdTnann-tvrc6Er8BBBgyY0</recordid><startdate>200905</startdate><enddate>200905</enddate><creator>Serpero, Laura D.</creator><creator>Kheirandish-Gozal, Leila</creator><creator>Dayyat, Ehab</creator><creator>Goldman, Julie L.</creator><creator>Kim, Jinkwan</creator><creator>Gozal, David</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley-Blackwell</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>200905</creationdate><title>A mixed cell culture model for assessment of proliferation in tonsillar tissues from children with obstructive sleep apnea or recurrent tonsillitis</title><author>Serpero, Laura D. ; Kheirandish-Gozal, Leila ; Dayyat, Ehab ; Goldman, Julie L. ; Kim, Jinkwan ; Gozal, David</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4867-fa34a1f7e4436f9566aefd7ea3ca711dda05b13c3a920841d9f0c51ea4e62e8e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Adenoidectomy</topic><topic>Adenoids - metabolism</topic><topic>Adenoids - pathology</topic><topic>adenotonsillectomy</topic><topic>Analysis of Variance</topic><topic>Biological and medical sciences</topic><topic>Cell Culture Techniques</topic><topic>Cell Proliferation</topic><topic>Child</topic><topic>Child, Preschool</topic><topic>cytokines</topic><topic>Cytokines - metabolism</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Female</topic><topic>Humans</topic><topic>Hypertrophy</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Obstructive sleep apnea</topic><topic>Otorhinolaryngology. Stomatology</topic><topic>Palatine Tonsil - metabolism</topic><topic>Palatine Tonsil - pathology</topic><topic>Pneumology</topic><topic>Recurrence</topic><topic>recurrent tonsillitis</topic><topic>Respiratory system : syndromes and miscellaneous diseases</topic><topic>Sleep Apnea, Obstructive - pathology</topic><topic>Sleep Apnea, Obstructive - surgery</topic><topic>tonsillar hypertrophy</topic><topic>Tonsillectomy</topic><topic>Tonsillitis - pathology</topic><topic>Tonsillitis - surgery</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Serpero, Laura D.</creatorcontrib><creatorcontrib>Kheirandish-Gozal, Leila</creatorcontrib><creatorcontrib>Dayyat, Ehab</creatorcontrib><creatorcontrib>Goldman, Julie L.</creatorcontrib><creatorcontrib>Kim, Jinkwan</creatorcontrib><creatorcontrib>Gozal, David</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Laryngoscope</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Serpero, Laura D.</au><au>Kheirandish-Gozal, Leila</au><au>Dayyat, Ehab</au><au>Goldman, Julie L.</au><au>Kim, Jinkwan</au><au>Gozal, David</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A mixed cell culture model for assessment of proliferation in tonsillar tissues from children with obstructive sleep apnea or recurrent tonsillitis</atitle><jtitle>The Laryngoscope</jtitle><addtitle>The Laryngoscope</addtitle><date>2009-05</date><risdate>2009</risdate><volume>119</volume><issue>5</issue><spage>1005</spage><epage>1010</epage><pages>1005-1010</pages><issn>0023-852X</issn><eissn>1531-4995</eissn><coden>LARYA8</coden><abstract>Background: Recurrent infective tonsillitis (RI) and obstructive sleep apnea (OSA) are the major indications for adenotonsillectomy (T&amp;A) in children. However, little is known on the determinants of lymphadenoid tissue proliferation in the pediatric upper airway. Objectives: 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&amp;A, were dissociated using standard methods. Whole cell tonsillar cultures were either maintained in normal medium or stimulated with lipopolysaccharide (25 μg/mL) and concanavalin A (10 μg/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 &lt; .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. Laryngoscope, 2009</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>19266584</pmid><doi>10.1002/lary.20147</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record>
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subjects Adenoidectomy
Adenoids - metabolism
Adenoids - pathology
adenotonsillectomy
Analysis of Variance
Biological and medical sciences
Cell Culture Techniques
Cell Proliferation
Child
Child, Preschool
cytokines
Cytokines - metabolism
Enzyme-Linked Immunosorbent Assay
Female
Humans
Hypertrophy
Male
Medical sciences
Obstructive sleep apnea
Otorhinolaryngology. Stomatology
Palatine Tonsil - metabolism
Palatine Tonsil - pathology
Pneumology
Recurrence
recurrent tonsillitis
Respiratory system : syndromes and miscellaneous diseases
Sleep Apnea, Obstructive - pathology
Sleep Apnea, Obstructive - surgery
tonsillar hypertrophy
Tonsillectomy
Tonsillitis - pathology
Tonsillitis - surgery
title A mixed cell culture model for assessment of proliferation in tonsillar tissues from children with obstructive sleep apnea or recurrent tonsillitis
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