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Role of ADAM10 and ADAM17 in CD16b Shedding Mediated by Different Stimulators.
MedLine Citation:
PMID:  22770404     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Objective To investigate the main proteinases responsible for CD16b shedding under different stimulators. Methods HEK293 cell line stably expressing CD16b was constructed by lentivirus system. The cell line was then overexpressed with a disintegrin and metalloproteinase 10 (ADAM10) or ADAM17, suppressed with short hairpin RNA of ADAM10 or ADAM17, and reconstituted with ADAM10 or ADAM17, respectively. After each treatment, the cell line was stimulated with ionomycin or phorbol 12-myristate- 13-acetate (PMA) for 12 hours. The soluble CD16b released from cell membrane was detected by immunoprecipition and immunoblot. Quantitation was then implemented to compare the amount of soluble CD16b in cell supernatant after stimulation. Results HEK293 cell line stably expressing CD16b was successfully established. When CD16b expressing cell line was overexpressed with ADAM10, shedding of CD16b was increased after stimulation with ionomycin but not PMA; when the cell line overexpressed with ADAM17, shedding of CD16b was increased after stimulation with PMA but not ionomycin. Similarly, when ADAM10 was suppressed by short hairpin RNA, CD16b shedding was decreased after stimulation with ionomycin; when ADAM17 was suppressed by short hairpin RNA, CD16b shedding was decreased after stimulation with PMA. The shedding of CD16b was increased again when CD16b expressing cell line was reconstituted with ADAM10 and stimulated by ionomycin or reconstituted with ADAM17 and stimulated by PMA. Conclusions Both ADAM10 and ADAM17 could shed CD16b, but they possess differed preferences. ADAM10 is the main sheddase under stimulation of ionomycin, while ADAM17 is the main sheddase under stimulation of PMA.
Authors:
Sha Guo; Min Peng; Qing Zhao; Wei Zhang
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Chinese medical sciences journal = Chung-kuo i hsüeh k'o hsüeh tsa chih / Chinese Academy of Medical Sciences     Volume:  27     ISSN:  1001-9294     ISO Abbreviation:  Chin. Med. Sci. J.     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-07-09     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9112559     Medline TA:  Chin Med Sci J     Country:  China    
Other Details:
Languages:  eng     Pagination:  73-9     Citation Subset:  IM    
Affiliation:
Department of Immunology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
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