JP7462864B2 - メンブレンヴェシクル - Google Patents
メンブレンヴェシクル Download PDFInfo
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- JP7462864B2 JP7462864B2 JP2019080136A JP2019080136A JP7462864B2 JP 7462864 B2 JP7462864 B2 JP 7462864B2 JP 2019080136 A JP2019080136 A JP 2019080136A JP 2019080136 A JP2019080136 A JP 2019080136A JP 7462864 B2 JP7462864 B2 JP 7462864B2
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- 102000004169 proteins and genes Human genes 0.000 claims description 17
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- 239000002158 endotoxin Substances 0.000 claims description 15
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- 241000605862 Porphyromonas gingivalis Species 0.000 claims description 7
- 229960005486 vaccine Drugs 0.000 description 16
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 8
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- XZKIHKMTEMTJQX-UHFFFAOYSA-N 4-Nitrophenyl Phosphate Chemical compound OP(O)(=O)OC1=CC=C([N+]([O-])=O)C=C1 XZKIHKMTEMTJQX-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Description
配列番号1記載のヌクレオチド配列からなるN末端モチーフ部分と、配列番号2記載のヌクレオチド配列からなるC末端モチーフ部分とにより、N末端とC末端領域とが付加されたGFPシーケンスの全ヌクレオチド配列(上流にプロモータ配列及び下流にターミネータ配列を含めて)をpTCBベクターへ導入したGFP発現ベクターを作成した(図2)。GFP発現ベクターの配列情報は配列番号3にて示される。
このGFP発現ベクターをエレクトロポレーション法にてPorphyromonas gingivalisへ導入した。0.7マイクログラム/ミリリットルの濃度のテトラサイクリンを含む寒天培地にてセレクションを行い、目的のプラスミドがPCR等により導入されていることを確認した(図3)。これにより菌体へのGFP発現ベクター導入の完了が確認された。
配列番号3にて示される配列からなるGFP発現ベクターを使用してマウスに経鼻免疫した。GFPタンパクを固相化したEnzyme-Linked ImmunoSorbent Assay (ELISA)を実施した。陰性標準プラスミド(空ベクター)を導入したPgのMV (Pg MV)及びGFP発現ベクターを導入したPgのMV (Pg MV -GFP)をマウスに免疫して得られた血清を、それぞれ0.02% Tween 20含有PBS にて200倍希釈したものを血清抗体液として、上記ELISAプレートを用い、通法通りELISAを行なった。二次抗体には、アルカリフォスファターゼ(ALP)標識二次抗体であるAP-labeled anti-マウスIgG (インビトロジェン社製)を、ALP基質パラニトロフェニルリン酸を使用して、吸光値A405にて発色をプレートリーダーで検出した。図4は、Enzyme-Linked ImmunoSorbent Assay (ELISA)によるPorphyromonas gingivalisのメンブレンヴェシクルで免疫したマウス血清におけるGFP抗体の発現の解析結果を示す。図4に示されるように、配列番号3にて示される配列からなるGFP発現ベクターを使用して経鼻免疫したマウスでは、コントロールと比較してGFP抗体の産生が有意に上昇していた。
配列番号2:C末モチーフ
配列番号3:ベクター
Claims (1)
- 9型分泌機構を有する細菌のMVであって、
前記細菌がPorphyromonas gingivalisであり、
前記MVが細胞外へ放出されたメンブレンヴェシクルであり、
外膜の構成部分であるリポ多糖に外来性タンパク質がアンカリングされており、前記外来性タンパク質が抗原性を有していることを特徴とする、MV。
Priority Applications (1)
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JP2019080136A JP7462864B2 (ja) | 2019-04-19 | 2019-04-19 | メンブレンヴェシクル |
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JP2019080136A JP7462864B2 (ja) | 2019-04-19 | 2019-04-19 | メンブレンヴェシクル |
Publications (2)
Publication Number | Publication Date |
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JP2020176221A JP2020176221A (ja) | 2020-10-29 |
JP7462864B2 true JP7462864B2 (ja) | 2024-04-08 |
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JP2019080136A Active JP7462864B2 (ja) | 2019-04-19 | 2019-04-19 | メンブレンヴェシクル |
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JP (1) | JP7462864B2 (ja) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018096007A2 (en) | 2016-11-25 | 2018-05-31 | Glaxosmithkline Biologicals S.A. | Immunogenic conjugates and use thereof |
JP2018521632A (ja) | 2015-06-02 | 2018-08-09 | デ スタート デル ネーデルランデン, ヴェルト. ドール デ ミニステル ヴァン ヴイダブリューエス ミニステリー ヴァン ボルクスゲツォントヘイト, ベルジーン エン シュポルトDe Staat Der Nederlanden, Vert. Door De Minister Van Vws Ministerie Van Volksgezondheid, Welzijn En Sport | グラム陰性外膜小胞における抗原の表面提示 |
-
2019
- 2019-04-19 JP JP2019080136A patent/JP7462864B2/ja active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2018521632A (ja) | 2015-06-02 | 2018-08-09 | デ スタート デル ネーデルランデン, ヴェルト. ドール デ ミニステル ヴァン ヴイダブリューエス ミニステリー ヴァン ボルクスゲツォントヘイト, ベルジーン エン シュポルトDe Staat Der Nederlanden, Vert. Door De Minister Van Vws Ministerie Van Volksgezondheid, Welzijn En Sport | グラム陰性外膜小胞における抗原の表面提示 |
WO2018096007A2 (en) | 2016-11-25 | 2018-05-31 | Glaxosmithkline Biologicals S.A. | Immunogenic conjugates and use thereof |
Non-Patent Citations (2)
Title |
---|
SCIENTIFIC REPORTS,2016年,Vol.6, Article No.24931,p.1-9 |
Vaccine,2016年,Vol.34,p.4626-4634 |
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JP2020176221A (ja) | 2020-10-29 |
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