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ID 47022
本文ファイル
著者
Hiragun, Takaaki 大学院医歯薬保健学研究院(医)
Ishii, Kaori
Hiragun, Makiko 病院(医)
Yanase, Yuhki 大学院医歯薬保健学研究科(医) 広大研究者総覧
Tanaka, Akio 大学院医歯薬保健学研究科(医) 広大研究者総覧
Hide, Michihiro 大学院医歯薬保健学研究科(医) 広大研究者総覧
キーワード
HRT
MGL_1304
Pichia pastoris
Sweat allergy
β-hexosaminidase release
NDC
医学
抄録(英)
Background: We previously identified MGL_1304 secreted by Malassezia globosa as a sweat antigen for patients with atopic dermatitis (AD) and cholinergic urticaria (ChU). However, purifying native MGL_1304 from human sweat or culture supernatant of M. globosa (sup-MGL_1304) is costly and timeconsuming. Moreover, recombinant MGL_1304 expressed by using Escherichia coli (TF-rMGL_1304) needs a large chaperon protein and lacks the original glycosylation of yeasts. Thus, we generated a recombinant MGL_1304 by Pichia pastoris (P-rMGL_1304) and investigated its characteristic features.
Methods: Recombinant MGL_1304 proteins expressed by E. coli and P. pastoris were generated. Properties of these recombinants and native antigens were compared by western blot analysis, histamine release tests (HRT) of patients with AD and ChU, and b-hexosaminidase release tests with RBL-48 cells. PrMGL_ 1304-specific IgE in sera of patients with AD were measured by sandwich ELISA.
Results: Western blot analysis revealed that IgE of patients with AD bound to all MGL_1304 recombinants and native antigens. The histamine releasing ability of P-rMGL_1304 was 100 times higher than that of TF-rMGL_1304, and was comparable to that of sup-MGL_1304. Degranulation rates of RBL-48 cells, sensitized with sera of patients with AD in response to the stimulation of P-rMGL_1304, were comparable to those of sup-MGL_1304, whereas those of TF-rMGL_1304 were relatively weak. The levels of P-rMGL_1304-specific IgE in sera of patients with AD were correlated with their disease severities.
Conclusions: P-rMGL_1304 has an antigenicity comparable to the native antigen, and is more useful than TF-rMGL_1304, especially in HRT and degranulation assay of RBL-48 cells.
内容記述
This work was supported in part by the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT/ JSPS KAKENHI, Grant numbers 19790789, 22591239, 25461669), Grant-in-Aid for Young Scientists (B), and Scientific Research (C); and Health and Labour Sciences Research Grant; A-STEP program of Japan Science and Technology Agency.
掲載誌名
Allergology International
64巻
3号
開始ページ
266
終了ページ
271
出版年月日
2015-07
出版者
Japanese Society of Allergology
ISSN
1323-8930
出版者DOI
PubMedID
言語
英語
NII資源タイプ
学術雑誌論文
広大資料タイプ
学術雑誌論文
DCMIタイプ
text
フォーマット
application/pdf
著者版フラグ
publisher
権利情報
Copyright © 2015, Japanese Society of Allergology. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/)
関連情報URL
部局名
医歯薬保健学研究科
病院