JP2005533855A - 病原性ウイルスからの別のリーディングフレームによりコードされる抗原 - Google Patents
病原性ウイルスからの別のリーディングフレームによりコードされる抗原 Download PDFInfo
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- JP2005533855A JP2005533855A JP2004523778A JP2004523778A JP2005533855A JP 2005533855 A JP2005533855 A JP 2005533855A JP 2004523778 A JP2004523778 A JP 2004523778A JP 2004523778 A JP2004523778 A JP 2004523778A JP 2005533855 A JP2005533855 A JP 2005533855A
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Abstract
Description
メチオニンアミノ酸残基から開始され、
抗原決定基を含み、かつ
7を超えるアミノ酸残基を含む
ことを特徴とするポリペプチド、および7を超えるアミノ酸を含む該ポリペプチドの断片を提供する。
実施例1:HCV
HCVを本発明のモデルウイルスとして用いた。しかしながら、本実施例に記載する原理はいかなるウイルスにも応用できる。
本発明により提供されるペプチドが免疫原性を有する可能性があるか否かを決定するため、HLA−A*0201対立遺伝子についてHCV1bからの3つのペプチドを選び、これらペプチドでHLA−A*0201トランスジェニックマウス(HHD)にワクチン接種した。
HLA−A*0201トランスジェニックマウス(1群当たり5匹)を以下のようにして皮下にてワクチン接種した:
(1)1371(HCV−H77 ncORF(1−3)11 TLWAGPLKV) + CpG1668
(2)1372(HCV−H77 ncORF(1−3)13 LLLQRWALV) + CpG1668
(3)1373(HCV−H77 ncORF(1−3)27 FMLGALLPI) + CpG1668
HLA−A*0201トランスジェニックマウス(1群当たり5匹)を以下のようにして皮下にてワクチン接種した:
(1)1445(HCV−1b ncORF(1−3)36 RLLQLKYCV) + CpG1668
(2)1447(HCV−1b ncORF(1−3)36 FLYLPLSFAV) + CpG1668
本発明によるncORFペプチドはtg−マウスにおいて免疫原性であるので、本発明のペプチドをHCV+患者からのPBLでELIspotアッセイにより分析した。
患者は1992年に慢性HCVに感染しており、1993年から1994年にIFN−アルファ単一療法で治療した。1996年に凍結した患者由来の末梢血単核細胞(PBMC)を解凍して以下のペプチドを用いてIFN−g Elispotアッセイを行った:
(1)1371(HCV−H77 ncORF(1−3)11 TLWAGPLKV)
(2)1372(HCV−H77 ncORF(1−3)13 LLLQRWALV)
(3)1373(HCV−H77 ncORF(1−3)27 FMLGALLPI)
(4)1006(HCV由来) MWNFISGIQYLAGLSTLPGN
(5)84(HCV由来) GYKVLVLNPSVAAT
(6)CMVpp65 NLVPMVATV
(7)インフルエンザAマトリックス(アミノ酸58〜67) GILGFVFTL
(Ipep1490;Ipep1491;Ipep1492;Ipep1493;Ipep1494;Ipep82)
HLA−A*0201トランスジェニックマウス(1群当たり5匹)を以下のようにして皮下にてワクチン接種した:
(1)1490(HCV−1b ncORF(4−6) KMLNRRVLWV) + CpG1668
(2)1491(HCV−1b ncORF(4−6) VLLMCQLPLV) + CpG1668
(3)1492(HCV−1b ncORF(4−6) MLNRRVLWVV) + CpG1668
(4)1493(HCV−1b ncORF(4−6) TILELEQSFV) + CpG1668
(5)1494(HCV−1b ncORF(4−6) KMMSPHAAV) + CpG1668
(6) 82(EBV、対照 GLCTLVAML) + CpG1668
ウイルスゲノムの異なるORF内に、可能なコードされたCTLエピトープを同定することができる;
これらORFのペプチドは、とりわけリーディングフレーム4〜6においても、tg−マウスで免疫原性である;
HCV+患者において陽性のELIspotの結果を与える、すなわちHCV感染において関連のある病理学的パラメーターである。
本実施例ではHIVのゲノムをその非コードORFに関して本発明に従って分析した。その結果を表6に示す。これらから、HIVワクチンの製造に抗原として用いるのが好ましい最小長の7アミノ酸残基を有するHIV−ncORFまたは7アミノ酸残基よりも長いHIV−ncORFを得ることができる。
本実施例では、実施例1でHCVエピトープについて記載したのと同様にしてHPVの優れた免疫特性を備えた可能なncORFエピトープを同定する。その結果を以下の表7に示す。
KLK/o−d(IC)13とともにインフルエンザAウイルスからのncORF由来ペプチドを用いたマウスのワクチン接種。ワクチン接種の7日後に特異的なT細胞応答を測定し、その後、被験動物を致死量のマウス適合インフルエンザAウイルス(×31)で攻撃する。生存を15日間モニターする。
マウス:C57B1/6(Harlan-Winkelmann、ドイツ)
ペプチド:p82(GLCTLVAML):EBV;HLA−A*0201;アミノ酸開始280からの対照ペプチド
p1574(IASNENMETM):インフルエンザ核タンパク質、アミノ酸開始365からの対照ペプチド
p1569(TMLYNKMEF):セグメント1、フレーム1、ORF1、アミノ酸開始569からのFlu ncORF由来ペプチド
p1600(SSIAAQDAL):セグメント3、フレーム6、ORF2、アミノ酸開始83からのFlu ncORF由来ペプチド
p1664(VTILNLALL):セグメント4、フレーム5、ORF6、アミノ酸開始9からのFlu ncORF由来ペプチド
投与量:100μg/ペプチド/マウス。
投与量:5ナノモル/マウス。
投与量:127ナノモル/マウス。
調合:270mMソルビトール/10mM Hepes
1.p1574 + KLK + o−d(IC)13
2.p1569 + KLK + o−d(IC)13
3.p1600 + KLK + o−d(IC)13
4.p1664 + KLK + o−d(IC)13
5.p1600 + p1569 + KLK + o−d(IC)13
1群および3群(ペプチドp1574およびp1600)の脾細胞は、各ペプチドで再刺激した後に特異的なスポットを示さない。2群および4群(p1569およびp1664)は再刺激後にIFN−γを特異的に放出する。5群は2つの個々のペプチド(混合物としてではなく、p1600およびp1569)でワクチン接種した。両ペプチドの混合物かまたはp1569のいずれかで再刺激すると特異的なサイトカイン放出が検出される。対照的に、p1600単独で再刺激するとIFN−γスポットは検出できない。これは3群(p1600単独)と一致するものである。
図5bは、致死量のマウス適合インフルエンザAウイルス×31で攻撃したマウスの生存率を示す。1群(p1574、H2−Dbについて報告された防御ペプチド)は攻撃した全マウスの30%を防御する。ペプチドp1569は全く防御を与えない。対照的に、ペプチドp1600およびp1664は、それぞれ攻撃した動物の50%および62%を防御する。動物を2つの異なるペプチドでワクチン接種すると(5群、ペプチドp1600およびp1664)、動物の70%が防御される。
Claims (27)
- 病原性ウイルスの別のリーディングフレームによりコードされるポリペプチドであって、
メチオニンアミノ酸残基から開始され、
抗原決定基を含み、かつ
7を超えるアミノ酸残基を含む
ことを特徴とするポリペプチド、および7を超えるアミノ酸を含む該ポリペプチドの断片。 - 該病原性ウイルスが、A型肝炎ウイルス(HAV)、B型肝炎ウイルス(HBV)、C型肝炎ウイルス(HCV)、D型肝炎ウイルス(HDV)、E型肝炎ウイルス(HEV)、F型肝炎ウイルス(HFV)、G型肝炎ウイルス(HGV)、ヒト免疫不全ウイルス、インフルエンザウイルス、口蹄疫ウイルス(FMDV)、エボラウイルス、HTLVI、HTLVII、SIV、パルボウイルス、パピローマウイルス、ロタウイルス、アデノウイルス、サイトメガロウイルス、ネコ免疫不全ウイルス(FIV)、エプスタインバーウイルス(EBV)、単純ヘルペスウイルス(HSV)、帯状疱疹ウイルス(HZV)、麻疹ウイルスおよび発癌ウイルスよりなる群から選ばれることを特徴とする、請求項1に記載のポリペプチドまたは断片。
- 少なくとも1の細胞障害性Tリンパ球(CTL)エピトープを含むことを特徴とする、請求項1または2に記載のポリペプチドまたは断片。
- A0201、A1、A24、A3、A31、B3501、B4403、B7、B8、とりわけA0201、またはそれらの混合物よりなる群から選ばれたHLA対立遺伝子に対する細胞障害性Tリンパ球(CTL)エピトープを含むことを特徴とする、請求項1ないし3のいずれかに記載のポリペプチドまたは断片。
- 少なくとも1のTヘルパー細胞エピトープを含むことを特徴とする、請求項1ないし4のいずれかに記載のポリペプチドまたは断片。
- DP、DQ、DRまたはそれらの混合物よりなる群から選ばれたHLA対立遺伝子に対するTヘルパー細胞エピトープを含むことを特徴とする、請求項1ないし5のいずれかに記載のポリペプチドまたは断片。
- 表2a)〜2n)(配列番号1〜822)に列挙した群から選ばれるポリペプチドまたは7を超えるアミノ酸を含む該ポリペプチドの断片。
- 表4a)〜4n)に列挙した群から選ばれた断片、好ましくは50またはそれ以上のスコアを有する断片、さらに好ましくは200を超えるスコアを有する断片、とりわけ500を超えるスコアを有する断片を含むかまたは該断片からなるポリペプチド。
- 表6に列挙され7を超えるアミノ酸残基を含むポリペプチド(配列番号823〜874)または7を超えるアミノ酸残基を含む該ポリペプチドの断片。
- 担体、とりわけ免疫修飾物質にコンジュゲートしていることを特徴とする、請求項1ないし9のいずれかに記載のポリペプチドまたは断片。
- ポリカチオン性物質、とりわけポリカチオン性ポリペプチド、および免疫修飾核酸、とりわけデオキシイノシンおよび/またはデオキシウリジン含有オリゴデオキシヌクレオチドよりなる群から選択される免疫修飾物質にコンジュゲートしていることを特徴とする、請求項1ないし10のいずれかに記載のポリペプチドまたは断片。
- 少なくとも1のT細胞エピトープを含むことを特徴とする、請求項1ないし11のいずれかに記載のポリペプチドまたは断片。
- 該病原性ウイルスの機能的なリーディングフレームの相補鎖を読みとる別のリーディングフレームによってコードされることを特徴とする、請求項1ないし12のいずれかに記載のポリペプチドまたは断片。
- 表4a、4c、4e、4g、4i、4kおよび4mに列挙され、50またはそれ以上のスコア、より好ましくは200を超えるスコア、とりわけ500を超えるスコアを有するペプチドの群から選択される少なくとも1のペプチドを含むことを特徴とする、請求項1ないし13のいずれかに記載のポリペプチドまたは断片。
- 治療剤として使用することを特徴とする、請求項1ないし14のいずれかに記載のポリペプチドまたは断片。
- N末端またはC末端の少なくとも一方に2ないし7のアミノ酸からなるテールを含み、該アミノ酸がF、I、L、A、Y、WまたはCから選ばれることを特徴とする、請求項1ないし15のいずれかに記載のポリペプチドまたは断片。
- N末端またはC末端の少なくとも一方に2ないし7のアミノ酸からなるテールを含み、該アミノ酸がEまたはDから選ばれることを特徴とする、請求項1ないし15のいずれかに記載のポリペプチドまたは断片。
- 50またはそれ以上のスコア、さらに好ましくは200を超えるスコア、とりわけ500を超えるスコアを有し、表7に列挙したペプチドの群から選ばれるペプチドを含むことを特徴とする、請求項1ないし17のいずれかに記載のポリペプチドまたは断片。
- 請求項1ないし18のいずれかに記載の1またはそれ以上のポリペプチドまたは断片を含む医薬組成物。
- ポリカチオン性物質、とりわけポリカチオン性ポリペプチド、および免疫修飾核酸、とりわけデオキシイノシンおよび/またはデオキシウリジン含有オリゴデオキシヌクレオチドよりなる群から好ましくは選択される免疫修飾物質をさらに含むことを特徴とする、請求項19に記載の医薬組成物。
- 病原性ウイルスの構造的または機能的ポリペプチドまたはその断片、とりわけ抗原決定基を含む構造的または機能的ポリペプチドまたはその断片をさらに含むことを特徴とする、請求項19または20に記載の医薬組成物。
- 請求項1ないし18のいずれかに記載の1またはそれ以上のポリペプチドまたは断片を単位投与当たり1ng〜1g、好ましくは100ng〜10mg、とりわけ10μg〜1mg含むことを特徴とする、請求項19ないし21のいずれかに記載の医薬組成物。
- ワクチンとして調合することを特徴とする、請求項19ないし22のいずれかに記載の医薬組成物。
- さらなる活性成分、とりわけ免疫強化サイトカイン、抗炎症物質、抗菌物質またはそれらの組み合わせを含むことを特徴とする、請求項19ないし23のいずれかに記載の医薬組成物。
- ポリカチオン性ペプチド、とりわけポリアルギニン、ポリリシンよりなる群から選ばれたポリカチオン性ポリマー、または抗菌ペプチド、とりわけカテリシジン由来の抗菌ペプチド、または成長ホルモン、とりわけヒト成長ホルモンをさらに含むことを特徴とする、請求項19ないし24のいずれかに記載の医薬組成物。
- 補助物質、とりわけ薬理学的に許容しうる担体、緩衝物質、安定化剤またはそれらの組み合わせをさらに含むことを特徴とする、請求項19ないし25のいずれかに記載の医薬組成物。
- 該病原性ウイルスの感染の治療または予防用医薬を製造するための請求項1ないし18のいずれかに記載のポリペプチドまたは断片の使用。
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Cited By (2)
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JP2021511054A (ja) * | 2018-01-24 | 2021-05-06 | ザ カウンシル オブ ザ クイーンズランド インスティテュート オブ メディカル リサーチ | Hpv免疫療法 |
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2003
- 2003-07-24 WO PCT/EP2003/008112 patent/WO2004011650A2/en active Application Filing
- 2003-07-24 AU AU2003254585A patent/AU2003254585A1/en not_active Abandoned
- 2003-07-24 CA CA002484941A patent/CA2484941A1/en not_active Abandoned
- 2003-07-24 CN CNA038097885A patent/CN1650012A/zh active Pending
- 2003-07-24 EP EP03771083A patent/EP1523557A2/en not_active Withdrawn
- 2003-07-24 JP JP2004523778A patent/JP2005533855A/ja active Pending
- 2003-07-24 US US10/512,790 patent/US7528223B2/en not_active Expired - Fee Related
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007105565A1 (ja) * | 2006-03-13 | 2007-09-20 | Keio University | インフルエンザ感染阻害ペプチド、インフルエンザウイルス感染阻害剤、リポソーム、インフルエンザ予防・治療剤 |
JP2021511054A (ja) * | 2018-01-24 | 2021-05-06 | ザ カウンシル オブ ザ クイーンズランド インスティテュート オブ メディカル リサーチ | Hpv免疫療法 |
Also Published As
Publication number | Publication date |
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CA2484941A1 (en) | 2004-02-05 |
US7528223B2 (en) | 2009-05-05 |
JP2009298821A (ja) | 2009-12-24 |
WO2004011650A2 (en) | 2004-02-05 |
US20070134262A1 (en) | 2007-06-14 |
AU2003254585A1 (en) | 2004-02-16 |
EP1523557A2 (en) | 2005-04-20 |
WO2004011650A3 (en) | 2004-06-24 |
CN1650012A (zh) | 2005-08-03 |
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