JPH05500599A - ヒト血小板膜糖タンパク質iiiaの多型性並びに診断及び治療用としてのその適用 - Google Patents
ヒト血小板膜糖タンパク質iiiaの多型性並びに診断及び治療用としてのその適用Info
- Publication number
- JPH05500599A JPH05500599A JP2506829A JP50682990A JPH05500599A JP H05500599 A JPH05500599 A JP H05500599A JP 2506829 A JP2506829 A JP 2506829A JP 50682990 A JP50682990 A JP 50682990A JP H05500599 A JPH05500599 A JP H05500599A
- Authority
- JP
- Japan
- Prior art keywords
- allele
- probe
- gpiiia
- molecule
- cdna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
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Abstract
Description
Claims (36)
- 1.GPIIIA遺伝子の一部分にハイブリダイズするオリゴヌクレオチドプロ ーブ分子であって、該部分がGPIIIA cDNAのヌクレオチド196を含 んでおり、該ヌクレオチド196がシトシンであるとき、該分子が該部分にハイ ブリダイズすることを特徴とする該オリゴヌクレオチドプローブ分子。
- 2.該分子が約20塩基長であることを特徴とする請求項31に記載のオリゴヌ クレオチドプローブ分子。
- 3.GPIIIA遺伝子の一部分にハイブリダイズするオリゴヌクレオチドプロ ーブ分子であって、該部分が該遺伝子のヌクレオチド196を含んでおり、該ヌ クレオチド196がチミジンであるとき、該分子が該部分にハイブリダイズする ことを特徴とする該オリゴヌクレオチドプローブ分子。
- 4.該分子が約20塩基長であることを特徴とする請求項33に記載のオリゴヌ クレオチドプローブ分子。
- 5.血小板P1Aアロ抗原を型別するためのキットであって、(a)血小板P1 Aの対立遺伝子を他の対立遺伝子から識別する標識化オリゴヌクレオチドプロー ブの溶液を収容する容器、(b)GPIIIAのP1A1対立遺伝子もしくはP 1A2対立遺伝子に識別可能に結合する抗体であって、(i)GPIIIAの少 なくともアミノ酸33をコードするヌクレオチド配列によりコードされるポリペ プチド分子を認識し、(ii)GPIIIAの該P1A1対立遺伝子又はP1A 2対立遺伝子のいずれかに結合する該抗体の溶液を収容する容器、又は(c)血 小板P1Aアロ抗原の対立遺伝子のヌクレオチド配列を他の対立遺伝子から識別 する切断部位を認識するエンドヌクレアーゼの溶液を収容する容器と、(d)該 GPIIIA cDNAのヌクレオチド196に対応するヌクレオチドを含むG PIIIA遺伝子もしくはGPIIIA cDNAの少なくとも一部分を含むD NAを増幅するための手段とを備える該キット。
- 6.ヒト血小板又は赤血球膜糖タンパク質の型別方法であって、(a)第1の個 体のヒト血小板又は赤血球mRNAからcDNAを合成する段階と、(b)該c DNAを増幅して増幅cDNAを作成する段階と、その後、(c)該増幅cDN Aを分析して血小板又は赤血球表現型を決定する段階とを含む該方法。
- 7.第2の個体のヒト血小板mRNAからcDNAを合成し、次いで該第2の個 体のcDNAで段階(b)及び(c)を繰り返し、その後、該アロ抗原表現型に 基づいて該第1及び第2の個体を識別する段階を更に含むことを特徴とする請求 項36に記載の方法。
- 8.段階(c)が、(i)血小板又は赤血球アロ抗原の対立遺伝子のヌクレオチ ド配列を他の対立遺伝子から識別する切断部位を認識する制限エンドヌクレアー ゼで該増幅cDNAを消化する段階と、(ii)該cDNAフラグメントを分析 して血小板又は赤血球表現型を決定する段階を含むことを特徴とする請求項36 に記載の方法。
- 9.(i)該cDNAがGPIIIA mRNAの少なくともヌクレオチド19 6をコードする配列を含み、(ii)該制限エンドヌクレアーゼがGPIIIA のヌクレオチド196を含むヌクレオチド配列を含むか又は該配列に隣接する制 限部位を認識することを特徴とする請求項38に記載の方法。
- 10.段階(c)が(i)P1A対立遺伝子のヌクレオチド配列を他のP1A対 立遺伝子から識別する標識化対立遺伝子特異的オリゴヌクレオチドプローブに該 cDNAフラグメントをハイブリダイズさせる段階と、その後、(ii)該cD NAフラグメントにハイブリダイズした該プローブを分析して該P1Aアロ抗原 表現型を決定する段階を含むことを特徴とする請求項38に記載の方法。
- 11.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 40に記載の方法。
- 12.段階(c)が、(i)第1のラベルが第2のラベルから識別可能であり且 つプローブが第1のP1A対立遺伝子を別のP1A対立遺伝子から識別するヌク レオチドに相互に隣接してハイブリダイズするように、プローブ対の第1のプロ ーブが第1のラベルで標識され且つプローブ対の第2のプローブが第2のラベル で標識された1対のオリゴヌクレオチドプローブに該増幅cDNAをハイブリダ イさせて構築物を作成する段階と、その後、(ii)該構築物を反応媒体中でリ ガーゼと反応させる段階と、その後、(iii)該反応媒体を分析して該第1の プローブ及び該第2のプローブを含む連結産物の存在を検出する段階とを含むこ とを特徴とする請求項38に記載の方法。
- 13.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 42に記載の方法。
- 14.(a)第1の個体からゲノムDNAを得る段階と、(b)該ゲノムDNA を分析して血小板P1Aアロ抗原表現型を決定する段階とを含む、血小板P1A 膜糖タンパク質の型別方法。
- 15.第2の個体からゲノムDNAを得る段階と、次いで該第2の個体のゲノム DNAで段階(b)を繰り返し、その後、該P1Aアロ抗原表現型に基づいて該 第1及び第2の個体を識別する段階を更に含むことを特徴とする請求項42に記 載の方法。
- 16.段階(b)が、(i)第1のP1A対立遺伝子のヌクレオチド配列を別の P1A対立遺伝子のヌクレオチド配列から識別する切断部位を認識する制限エン ドヌクレアーゼで該ゲノムDNAを消化してDNAフラグメントを作成する段階 と、その後、(ii)該DNAフラグメントを標識化プローブで分析して該P1 Aアロ抗原表現型を決定する段階を含むことを特徴とする請求項44に記載の方 法。
- 17.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 46に記載の方法。
- 18.段階(b)が、(i)該ゲノムDNAを制限エンドヌクレアーゼで消化し てDNAフラグメントを作成する段階と、その後、 (ii)血小板P1Aアロ 抗原の対立遺伝子のヌクレオチド配列を他の対立遺伝子から識別する標識化対立 遺伝子特異的オリゴヌクレオチドプローブに該DNAフラグメントをハイブリダ イズさせる段階と、その後、(iii)該DNAフラグメントにハイブリダイズ した該プローブを分析して該P1Aアロ抗原表現型を決定する段階を含むことを 特徴とする請求項44に記載の方法。
- 19.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 48に記載の方法。
- 20.段階(b)が、(i)第1のラベルが第2のラベルから識別可能であり且 つプローブ対がP1A対立遺伝子を別のP1A対立遺伝子から識別する該ゲノム DNAのヌクレオチドに相互に隣接してハイブリダイズするように、プローブ対 の第1のプローブが第1のラベルで標識され且つプローブ対の第2のプローブが 第2のラベルで標識された1対のオリゴヌクレオチドプローブに該ゲノムDNA をハイブリダイズさせて構築物を作成する段階と、その後、(ii)該構築物を 反応媒体中でリガーゼと反応させる段階と、その後、(iii)該反応媒体を分 析して該第1のプローブ及び該第2のプローブを含む連結産物の存在を検出する 段階とを含むことを特徴とする請求項44に記載の方法。
- 21.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 50に記載の方法。
- 22.GPIIIAに関する血小板の型別方法であって、(a)第1の個体から ゲノムDNAを得る段階と、(b)該ゲノムDNAを増幅して増幅ゲノムDNA を作成する段階と、(c)該増幅ゲノムDNAを分析して血小板P1Aアロ抗原 表現型を決定する段階とを含む該方法。
- 23.第2の個体からゲノムDNAを得る段階と、次いで該第2の個体のゲノム DNAで段階(b)及び(c)を繰り返し、その後、該アロ抗原表現型に基づい て該第1及び第2の個体を識別する段階を更に含むことを特徴とする請求項52 に記載の方法。
- 24.段階(C)が、 (i)第1のP1A対立遺伝子のヌクレオチド配列を別 のP1A対立遺伝子から識別する切断部位を認識する制限エンドヌクレアーゼで 該増幅ゲノムDNAを消化してDNAフラグメントを作成する段階と、その後、 (ii)該DNAフラグメントを分析して該アロ抗原表現型を決定する段階を含 むことを特徴とする請求項52に記載の方法。
- 25.段階(c)が、(i)P1A対立遺伝子のヌクレオチド配列を他のP1 A対立遺伝子のヌクレオチド配列から識別する標識化対立遺伝子特異的オリゴヌ クレオチドプローブに該増幅ゲノムDNAをハイブリダイズさせる段階と、その 後、(ii)該増幅ゲノムDNAフラグメントにハイブリダイズした該プローブ を分析して該アロ抗原表現型を決定する段階を含むことを特徴とする請求項52 に記載の方法。
- 26.該P1A対立遺伝子がP1A2対立遺伝子であることを特徴とする請求項 55に記載の方法。
- 27.段階(c)が、(i)第1のラベルが第2のラベルから識別可能であり且 つプローブがP1A対立遺伝子を別のP1A対立遺伝子から識別するヌクレオチ ドに相互に隣接してハイブリダイズするように、プローブ対の第1のプローブが 第1のラベルで標識され且つプローブ対の第2のプローブが第2のラベルで標識 された1対のオリゴヌクレオチドプローブに該増幅ゲノムDNAをハイブリダイ ズさせて構築物を形成する段階と、その後、(ii)該構築物を反応媒体中でリ ガーゼと反応させる段階と、その後、(iii)該反応媒体を分析して該第1の プローブ及び該 第2のプローブを含む連結産物の存在を検出する段階とを 含 むことを特徴とする請求項52に記載の方法。
- 28.該P1A対立遺伝子がP1A2対立遺伝子であること を特徴とする請 求項57に記載の方法。
- 29.GPIIIAのフラグメントに対応するアミノ酸配列を含むポリペプチ ド分子であって、該フラグメントがGPIIIAのアミノ酸33を含んでおり、 該分子がGPIIIA以外のものであることを特徴とする該ポリペプチド分子。
- 30.該分子が免疫原性であることを特徴とする請求項59に記載のポリペプチ ド分子。
- 31.GPIIIAのP1A1及びP1A2形を識別する抗体。
- 32.GPIIIAの別のP1A形よりも優先的にGPIIIAのあるP1A形 に結合する抗体の製造方法であって、(a)請求項59に記載のポリペプチドを 含む抗原分子で哺乳動物を免疫感作する段階と、その後、(b)該哺乳動物から リンパ球を取り出す段階と、(c)該リンパ球を哺乳動物ミエローマ細胞に融合 させてハイブリドーマ細胞を作成する段階と、(d)該ハイブリドーマ細胞を培 養する段階と、その後、(e)GPIIIAのP1A1及びP1A2 形を識別 するモノクローナル抗体を分泌するハイブリドー マ細胞を選択、単離及びクロ ーニングする段階とを含む該 方法。
- 33.輸血後紫斑病又は新生児同種免疫血小板減少症の治療方法であって、薬 理的に有効な濃度のポリペプチド及び生理学的に適合可能なキャリヤーを配合し てなる調合物を第1の個体に投与する段階を含み、該第1の個体が(i)輸血後 紫斑病患者又は新生児同種免疫血小板減少症を発病する危険のある胎児の母親で あり且つ(ii)抗P1A1又は抗P1A2抗体を有しており、該ポリペプチド が抗P1A1抗体及び抗P1A2抗体から構成される群から選択される抗体に結 合することを特徴とする該方法。
- 34.GPIIIA遺伝子の一部分に対応するヌクレオチド配列を含む単離DN A分子であって、該部分がヌクレオチド196を含んでおり、該分子がGPII IA遺伝子に一致しないことを特徴とする該単離DNA分子。
- 35.ヌクレオチド196がシトシンであることを特徴とする請求項64に記載 の単離DNA分子。
- 36.ヌクレオチド196がチミジンであることを特徴とする請求項64に記載 の単離DNA分子。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US07/343,827 US5091302A (en) | 1989-04-27 | 1989-04-27 | Polymorphism of human platelet membrane glycoprotein iiia and diagnostic and therapeutic applications thereof |
US343,827 | 1989-04-27 |
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JPH05500599A true JPH05500599A (ja) | 1993-02-12 |
JP3221679B2 JP3221679B2 (ja) | 2001-10-22 |
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US (3) | US5091302A (ja) |
EP (1) | EP0477201B1 (ja) |
JP (1) | JP3221679B2 (ja) |
AT (1) | ATE161055T1 (ja) |
AU (1) | AU5551690A (ja) |
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WO (1) | WO1990012593A1 (ja) |
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US4351901A (en) * | 1980-03-24 | 1982-09-28 | Cetus Corporation | Method for single nucleotide alteration |
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CA1338457C (en) * | 1986-08-22 | 1996-07-16 | Henry A. Erlich | Purified thermostable enzyme |
US5091302A (en) * | 1989-04-27 | 1992-02-25 | The Blood Center Of Southeastern Wisconsin, Inc. | Polymorphism of human platelet membrane glycoprotein iiia and diagnostic and therapeutic applications thereof |
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- 1989-04-27 US US07/343,827 patent/US5091302A/en not_active Expired - Lifetime
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1990
- 1990-04-23 AT AT90908017T patent/ATE161055T1/de not_active IP Right Cessation
- 1990-04-23 JP JP50682990A patent/JP3221679B2/ja not_active Expired - Lifetime
- 1990-04-23 AU AU55516/90A patent/AU5551690A/en not_active Abandoned
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- 1990-04-23 EP EP90908017A patent/EP0477201B1/en not_active Expired - Lifetime
- 1990-04-23 WO PCT/US1990/002104 patent/WO1990012593A1/en active IP Right Grant
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1991
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1995
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EP0477201B1 (en) | 1997-12-10 |
ATE161055T1 (de) | 1997-12-15 |
US5670337A (en) | 1997-09-23 |
JP3221679B2 (ja) | 2001-10-22 |
DE69031804D1 (de) | 1998-01-22 |
AU5551690A (en) | 1990-11-16 |
US5391714A (en) | 1995-02-21 |
EP0477201A1 (en) | 1992-04-01 |
US5091302A (en) | 1992-02-25 |
WO1990012593A1 (en) | 1990-11-01 |
DE69031804T2 (de) | 1998-04-02 |
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