JPH10507627A - Tリンパ球のインビトロ増殖の急速拡大方法(「rem」) - Google Patents
Tリンパ球のインビトロ増殖の急速拡大方法(「rem」)Info
- Publication number
- JPH10507627A JPH10507627A JP8508988A JP50898896A JPH10507627A JP H10507627 A JPH10507627 A JP H10507627A JP 8508988 A JP8508988 A JP 8508988A JP 50898896 A JP50898896 A JP 50898896A JP H10507627 A JPH10507627 A JP H10507627A
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- Prior art keywords
- cells
- cell
- lymphocytes
- culture
- rapid expansion
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- 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.)
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Abstract
Description
Claims (1)
- 【特許請求の範囲】 1. インビトロで培養培地中において初期Tリンパ球集団を急速に拡大させる 方法であって、以下の工程:初期Tリンパ球集団をインビトロで培養培地に添加 する工程;不釣り合いに多数の非分裂末梢血単核細胞(PBMC)を、フィーダ細胞 として該培養培地に添加し、得られる細胞集団が、拡大されるべき初期集団中に Tリンパ球の各々について少なくとも約40個のPBMCフィーダ細胞を含むようにす る工程;および該培養物をインキュベートする工程;を包含する、方法。 2. 前記初期Tリンパ球集団が、少なくとも1種のヒトCD8+抗原特異的細胞 傷害性Tリンパ球を含む、請求項1に記載の急速拡大方法。 3. 前記初期Tリンパ球集団が、少なくとも1種のヒトCD4+抗原特異的ヘル パーTリンパ球を含む、請求項1に記載の急速拡大方法。 4. 前記非分裂フィーダ細胞が、γ線照射PBMCフィーダ細胞を含む、請求項1 に記載の急速拡大方法。 5. PBMCフィーダ細胞の初期Tリンパ球に対する比が、少なくとも約200:1 である、請求項1に記載の急速拡大方法。 6. PBMCフィーダ細胞の初期Tリンパ球に対する比が、約400:1と約800:1 の間である、請求項5に記載の急速拡大方法。 7. 非分裂EBV形質転換リンパ芽球様細胞(LCL)をフィーダ細胞として添加す る工程をさらに包含する、請求項1に記載の急速拡大方法。 8. LCLフィーダ細胞の初期Tリンパ球に対する比が、少なくとも約10:1で ある、請求項7に記載の急速拡大方法。 9. LCLフィーダ細胞の初期Tリンパ球に対する比が、約50:1と約200:1の 間である、請求項8に記載の急速拡大方法。 10. 前記培養培地に抗CD3モノクローナル抗体を添加する工程をさらに包含 する、請求項1に記載の急速拡大方法。 11. 抗CD3モノクローナル抗体の濃度が、少なくとも約0.5ng/mlである、請 求項10に記載の急速拡大方法。 12. 抗CD3モノクローナル抗体の濃度が、少なくとも約1.0ng/mlである、請 求項11に記載の急速拡大方法。 13. 前記培養培地にIL-2を添加する工程をさらに包含する、請求項1に記載 の急速拡大方法。 14. IL-2の濃度が、少なくとも約10単位/mlである、請求項13に記載の急 速拡大方法。 15. 前記インキュベーションを少なくとも約9日間続け、かつ前記培養培地 にIL-2を添加する工程を3〜5日の間隔毎に繰り返す、請求項14に記載の急速 拡大方法。 16. ヒトT細胞に遺伝子形質導入する方法であって、以下の工程: 初期Tリンパ球集団をインビトロで培養培地に添加する工程; 不釣り合いに多数の非分裂末梢血単核細胞(PBMC)を、フィーダ細胞として該 培養培地に添加し、得られる細胞集団が、拡大されるべき初期集団中にTリンパ 球の各々について少なくとも約40個のPBMCフィーダ細胞を含むようにする工程; 該培養物をインキュベートする工程;および 該培養培地にベクターを添加する工程; を包含する、方法。 17. 非分裂EBV形質転換リンパ芽球様細胞(LCL)をさらなるフィーダ細胞と して添加する工程をさらに包含する、請求項16に記載の遺伝子形質導入方法。 18. 前記ベクターが、Tリンパ球を阻害する阻害化合物に対する耐性を付与 する選択マーカーを含むレトロウイルスベクターであり、かつ以下の工程: 該レトロウイルスベクターの添加後に少なくとも1日、前記培養物のインキュ ベーションを継続する工程;および 該継続されたインキュベーション工程の後に前記培養培地に該阻害化合物を添 加する工程; をさらに包含する、請求項17に記載の遺伝子形質導入方法。 19. 前記レトロウイルスベクターが、ポジティブ選択マーカーおよびネガテ ィブ選択マーカーの両方を含む、請求項18に記載の遺伝子形質導入方法。 20. 前記レトロウイルスベクターが、ハイグロマイシンホスホトランスフェ ラーゼ遺伝子および単純ヘルペスウイルスのチミジンキナーゼ遺伝子を融合遺伝 子としてコードする、請求項19に記載の遺伝子形質導入方法。
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US29993094A | 1994-08-31 | 1994-08-31 | |
US08/299,930 | 1994-08-31 | ||
US08/317,100 | 1994-10-03 | ||
US08/317,100 US5827642A (en) | 1994-08-31 | 1994-10-03 | Rapid expansion method ("REM") for in vitro propagation of T lymphocytes |
PCT/US1995/011182 WO1996006929A2 (en) | 1994-08-31 | 1995-08-31 | Rapid expansion method ('rem') for in vitro propagation of t lymphocytes |
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JPH10507627A true JPH10507627A (ja) | 1998-07-28 |
JP4001912B2 JP4001912B2 (ja) | 2007-10-31 |
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JP50898896A Expired - Fee Related JP4001912B2 (ja) | 1994-08-31 | 1995-08-31 | Tリンパ球のインビトロ増殖の急速拡大方法(「rem」) |
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US (2) | US5827642A (ja) |
EP (1) | EP0778887A1 (ja) |
JP (1) | JP4001912B2 (ja) |
AU (1) | AU707880B2 (ja) |
CA (1) | CA2198633C (ja) |
WO (1) | WO1996006929A2 (ja) |
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EP0348413A1 (en) * | 1987-03-11 | 1990-01-03 | The Children's Hospital, Incorporated | Method for the generation of antigen-specific t cell lines and therapeutic use thereof |
US5057423A (en) * | 1987-12-18 | 1991-10-15 | University Of Pittsburgh | Method for the preparation of pure LAK-active lymphocytes |
US5399346A (en) * | 1989-06-14 | 1995-03-21 | The United States Of America As Represented By The Department Of Health And Human Services | Gene therapy |
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DE69128037T2 (de) * | 1990-11-13 | 1998-05-07 | Immunex Corp | Bifunktionelle wählbare fusionsgene |
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- 1995-08-31 JP JP50898896A patent/JP4001912B2/ja not_active Expired - Fee Related
- 1995-08-31 CA CA2198633A patent/CA2198633C/en not_active Expired - Lifetime
- 1995-08-31 EP EP95931690A patent/EP0778887A1/en not_active Withdrawn
- 1995-08-31 WO PCT/US1995/011182 patent/WO1996006929A2/en not_active Application Discontinuation
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CA2198633C (en) | 2010-03-30 |
WO1996006929A3 (en) | 1996-03-28 |
US6040177A (en) | 2000-03-21 |
AU707880B2 (en) | 1999-07-22 |
WO1996006929A2 (en) | 1996-03-07 |
EP0778887A1 (en) | 1997-06-18 |
JP4001912B2 (ja) | 2007-10-31 |
AU3503195A (en) | 1996-03-22 |
CA2198633A1 (en) | 1996-03-07 |
US5827642A (en) | 1998-10-27 |
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