JP2014152125A - IgA分泌促進剤 - Google Patents
IgA分泌促進剤 Download PDFInfo
- Publication number
- JP2014152125A JP2014152125A JP2013021411A JP2013021411A JP2014152125A JP 2014152125 A JP2014152125 A JP 2014152125A JP 2013021411 A JP2013021411 A JP 2013021411A JP 2013021411 A JP2013021411 A JP 2013021411A JP 2014152125 A JP2014152125 A JP 2014152125A
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- Prior art keywords
- iga
- iga secretion
- indigestible dextrin
- secretion promoter
- indigestible
- Prior art date
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Abstract
【解決手段】難消化性デキストリンを経口投与することにより有意なIgA分泌促進作用が認められた。
【選択図】なし
Description
マウスによる飼育実験の結果、水溶性食物繊維の一種である難消化性デキストリンを配合した飼料で飼育したマウスでは、腸管内に分泌されたIgAおよび糞便中のIgA量が難消化性デキストリンの用量に依存して増加し、新規IgA分泌促進剤として有効であることが明らかになった。
よって、難消化性デキストリンが粘膜免疫の賦活作用を有することが明らかになった。
すなわち本発明は、難消化性デキストリンを有効成分とするIgA分泌促進剤である。
0.1Mリン酸二水素ナトリウム(pH9.0)で10μg/mlに希釈したヤギ抗マウスIgG F(ab')2抗体(SIGMA)を96ウェルマイクロタイタープレート(Nunc)に50μl/well添加し、4℃で一晩インキュベートして抗体をプレートに吸着させた。0.05% Tween-20含有Phosphate buffered saline (PBST)でウェルを3回洗浄後、1%BSA-PBSを100μl添加して室温で2時間インキュベートしブロッキングを行った。PBSTで3回洗浄後、4℃、300G、10分間の遠心分離により得られた培養7日目のパイエル板(PP)細胞の培養上清を1%BSA-PBSTで1/50に希釈し、50μl添加した。腸内容物抽出液は同様に1/2000に希釈して50μl添加した。標準液は精製マウス骨髄腫IgA抗体(Kappa)(Bethyl Laboratories, Montgomery, TX)を1%BSA-PBSTで200ng/mlに希釈し、1/2ずつ段階希釈した。これを50μlウェルに添加し、検量線作成のための標準液として用いた。PBSTで4回洗浄後、1%BSA-PBSTで1/2000に希釈したアルカリフォスファターゼ標識ヤギ抗マウスIgA(α chain specific)抗体(Southern Biotech, Birmingham, AL)を50μl添加し、室温で2時間インキュベートした。PBSTで8回洗浄後、4-ニトロフェニルリン酸二ナトリウム(東京化成工業, 東京)をジエタノールアミン緩衝液に1mg/mlの濃度で溶解し50μl添加した。遮光したプレートを37℃で20〜30分インキュベート後、Microplate Reader Model 550(Bio-Rad Laboratories, Alfred Nobel Drive Hercules, CA)で405nmの吸光値を測定し、解析はMicro Plate Manager III(Bio-Rad Laboratories)を用いて行った。
0.1Mリン酸二水素ナトリウム(pH9.0)で2μg/mlに希釈したラット抗マウスIL-12(p40/p70)抗体(BD pharmigen, San Diego, CA, USA)を96ウェルマイクロタイタープレート(Nunc)に50μl/well添加し、4℃で一晩インキュベートして抗体をプレートに吸着させた。PBSTでウェルを3回洗浄後、1%BSA-PBSを100μl添加して室温で2時間インキュベートしブロッキングを行った。PBSTで3回洗浄後、4℃、300G、10分間の遠心分離により得られた培養24時間後のPP細胞の培養上清を50μl添加した。標準品はリコンビナントマウスIL-12p40 (BD pharmigen, San Diego, CA, USA)を 1%BSA-PBSTで4000pg/mlに希釈し、1/2ずつ段階希釈した。これを50μl添加し、検量線作成のための標準液として用いた。PBSTで4回洗浄後、1%BSA-PBSTで2μg/mlに希釈したビオチン標識ラット抗マウスIL-12(p40/p70)抗体(BD pharmigen, San Diego, CA, USA)を50μl添加し、室温で2時間インキュベートした。PBSTで6回洗浄後、アルカリフォスファターゼ標識ストレプトアビジン(Zymed, San Francisco, CA)を1%BSA-PBSTで0.6μg/mlに希釈して50μl添加し、室温で2時間インキュベートした。PBSTで6回洗浄後、4-ニトロフェニルリン酸二ナトリウム(東京化成工業, 東京)をジエタノールアミン緩衝液に1mg/mlの濃度で溶解し50μl添加した。遮光したプレートを37℃で約120分インキュベート後、Microplate Reader Model 550(Bio-Rad Laboratories, Alfred Nobel Drive Hercules, CA)で405nmの吸光値を測定し、解析はMicro Plate Manager III(Bio-Rad Laboratories)を用いて行った。
細胞培養液中のIgAは、LPS及びconAを添加した際に難消化性デキストリン摂取群では増加しており、難消化性デキストリンはパイエル板のIgA産生能を亢進させることが明らかとなった(図3)。
IL-12に関しても、分泌活性剤を添加した場合は、難消化性デキストリン摂取群でIL-12量が増加した(図4)。IL-12はNK細胞に対する著明な活性化作用を特徴とするサイトカインである。IL-12はB細胞および単球系細胞より産生され、T細胞やNK細胞に対して細胞増殖の促進、細胞傷害活性誘導、IFN-γ産生誘導、LAK細胞誘導などの作用を示す。こうした細胞性免疫機能への役割から、IL-12には感染防御や免疫不全症の改善における臨床応用が期待されている。例えば、HIV感染患者の末梢血リンパ球におけるIL-12産生、IFN-γ産生あるいはNK細胞活性はいずれも有意に低下しているが、IL-12の投与によってこれらを健常者と同程度までに増強することが知られている。よって、難消化性デキストリンの摂取によりIgA分泌能及びIL-12産生能が高まることは、難消化性デキストリンが粘膜免疫を賦活することを示している。
Claims (4)
- 難消化性デキストリンを有効成分とするIgA分泌促進剤。
- 難消化性デキストリンの難消化性成分が45質量%以上である請求項1記載のIgA分泌促進剤。
- IgA分泌促進剤が経口投与形態の剤形に調製されている請求項1又は2に記載のIgA分泌促進剤。
- 請求項1〜3のいずれか一項に記載のIgA分泌促進剤を含む粘膜免疫賦活剤、抗感染症剤又は抗アレルギー剤。
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JP2013021411A JP6160011B2 (ja) | 2013-02-06 | 2013-02-06 | IgA分泌促進剤 |
TW102113294A TWI598102B (zh) | 2013-02-06 | 2013-04-15 | 使用難消化性糊精在製備IgA分泌促進劑、黏膜免疫活化劑、抗感染症劑及抗過敏劑的用途 |
KR1020130040848A KR20140100380A (ko) | 2013-02-06 | 2013-04-15 | IgA 분비 촉진제 |
US13/871,517 US20140220080A1 (en) | 2013-02-06 | 2013-04-26 | IgA SECRETION PROMOTER |
KR1020190048192A KR101985644B1 (ko) | 2013-02-06 | 2019-04-25 | 면역 증진제 및 이를 포함하는 면역 개선용 음식품 조성물 |
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EP1629727A1 (en) * | 2004-08-24 | 2006-03-01 | N.V. Nutricia | Composition containing fermentable polysaccharides |
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Cited By (4)
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WO2019082897A1 (ja) | 2017-10-26 | 2019-05-02 | 株式会社ダイセル | 腸管免疫亢進剤、食品、及び医薬 |
KR20200051016A (ko) | 2017-10-26 | 2020-05-12 | 주식회사 다이셀 | 장관 면역 항진제, 식품, 및 의약 |
KR20230074280A (ko) | 2017-10-26 | 2023-05-26 | 주식회사 다이셀 | 장관 면역 항진제, 식품, 및 의약 |
US11896043B2 (en) | 2017-10-26 | 2024-02-13 | Daicel Corporation | Intestinal immune-enhancing agent, food product, and medicament |
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Free format text: JAPANESE INTERMEDIATE CODE: R250 |