JP2016504913A - 培養物中のウイルス収量を増加させるためのiNOS阻害剤の使用 - Google Patents
培養物中のウイルス収量を増加させるためのiNOS阻害剤の使用 Download PDFInfo
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Abstract
Description
(a)V27細胞にHSV−1 d27.1ベクターを感染させること;および
(b)アウリントリカルボン酸、バルプロ酸またはデキサメタゾンを含む細胞培養物中で感染したV27細胞を培養すること
を含む、HSV−1 d27.1ベクターを培養するための方法に関する。
本発明を記載する際に、以下の用語が用いられ、以下に示されるように定義されることが意図される。
本発明を詳細に記載する前に、本発明が、そのようなものとして勿論変化してもよい特定の製剤またはプロセスパラメータに限定されないことが理解されるべきである。また、本明細書で用いられる用語は、本発明の特定の実施形態のみを記載するためのものであり、限定を意図するものではない。
以下は、本発明を実行するための特定の実施形態の例である。実施例は、例示目的のみで提供されるものであり、いかなる意味でも本発明の範囲を限定することを意図するものではない。
細胞
Vero由来V27細胞(Riceら、J.Virol.(1989)63:3399〜3407頁)およびヒト胚性腎細胞(HEK)由来293細胞(Grahamら、J.Gen.Virol.(1977)36:59〜74頁)を、Applied Genetic Technologies Corporation(AGTC、Alachua、FL)から得て、Vero細胞およびHeLa細胞を、American Type Culture Collection(ATCC、Manassas、VA)から購入した。全ての細胞を、10%ウシ胎仔血清(FBS;HyClone)およびV27細胞についてはゲネチシン(50mg/ml;Invitrogen)または他の細胞については1%ペニシリン/ストレプトマイシン(Cellgro Mediatech、Manassas、VA)のいずれかを含有するDulbecco改変Eagle培地(DMEM;HyClone、South Logan、UT)中で維持した。
wtHSV−1 KOS株およびwtHSV−1 KOS株のICP27欠損誘導体:ベクターd27−1(RiceおよびKnipe、J.Virol.(1990)64:1704〜1715頁)、rHSV−rep2/cap2およびrHSV−EGFP(Kangら、Gene Ther.(2009)16:229〜239頁)ならびにその産生株ICP27コンプリメントV27細胞系を、AGTC(Alachua、FL)から得た。Advanced Biotechnologies Inc.(ABI、Columbia MD)から購入したwtHSV−1 MacIntyre株およびwtHSV−1 KOS株を、Vero、293またはHeLa細胞系中で増殖させた。感染性ベクター粒子を、培養上清を回収することにより感染の72h後に収穫した。DNase耐性粒子/ml(DRP/ml)中のHSV−1ストックの力価を、Taqmanアッセイにより決定した。粗培養培地内のウイルスゲノムを、37℃で60minのDNaseI(最終50U/ml;Promega)の存在下での処理、次に50℃で60minのプロテイナーゼK(Invitrogen)消化(1U/ml)、次いで、95℃で30minの変性により定量した。線状化されたプラスミドpZero195UL36(AGTC,Inc.、Alachua、FLから得た)を用いて、標準曲線を作製した。プライマー−プローブセットは、ベクターゲノムUL36配列(HSV−UL36 F:5’−GTTGGTTATGGGGGAGTGTGG(配列番号1);HSV−UL36 R:5’−TCCTTGTCTGGGGTGTCTTCG(配列番号2);HSV−UL36プローブ:5’−6FAM−CGACGAAGACTCCGACGCCACCTC−TAMRA(配列番号3))に特異的であった。PCR産物の増幅を、以下のサイクリングパラメータ:50℃で2minの1サイクル、95℃で10minの1サイクル;95℃で15sec、および60℃で60secの40サイクルを用いて達成した。
ATA(Sigma−A1895アウリントリカルボン酸実用等級、85%以上(滴定)、粉末)ストック溶液を、100mM重炭酸ナトリウム水溶液中500μM濃度として作製した。ATAストック溶液を、DMEM±10%ウシ胎仔血清(FBS、Hyclone、Waltham、MA)中でさらに希釈し、12〜60μM ATAの範囲の濃度(8.5〜21μg ATA/ml)にした。0.15の感染多重度(MOI=0.15)でのHSV−1感染(典型的には、6ウェルプレート中で6x105細胞)を、1〜2h、総最終培地容量の40%(2/5vol)中で行い、残りの培地(総最終容量の60%または3/5)を希釈工程中に添加した。次いで、細胞を72時間インキュベートし、上清を収穫して、1ミリリットルあたりの形成単位(PFU/ml)およびDRP/ml力価アッセイを行った。
デキサメタゾン(Sigma−D4902)を、無水アルコール中で2mg/mlに溶解した。これをDMEMで希釈して、1M濃度を達成し、−20℃で保存した。
バルプロ酸(Sigma−P4543)を、水中で1M濃度となるように溶解した。V27細胞を、6x105細胞/ウェルでの感染の前日に6ウェルプレート中に播種した。1Mバルプロ酸をDMEM−10%FBS 1mlに加え、5μMの濃度を達成した。これをよくボルテックスし、ウェルに添加した。プレートを6時間インキュベートし、吸引し、感染性HSV−1 d27−1ストックを、DMEM(添加物なし)1mlあたりMOI0.15(=細胞播種密度x0.15/pfu力価のHSVストック)で添加した。
293細胞(2.5x106個)に、Kangら、Gene Ther(2009)16:229〜239頁により記載されたようにrHSV−rep2/cap2とrHSV−EGFPベクターの両方を同時に感染させた。感染後2〜4hで、感染性培地をDMEM+10%FBS等価物と交換し、感染前培養容量を倍化した。収穫時に、細胞ペレットを−80℃で凍結した。DRP力価を、96ウェルブロックサーモサイクラー(Applied Biosystems;7500 Real Time PCR system)中でのリアルタイムポリメラーゼ連鎖反応(qPCR)により定量した。粗試料を3サイクルの凍結および解凍にかけた後、ベンゾナーゼ(250U/ml)、2mM MgCl2、1%最終濃度タンパク質等級Tween80(Calbiochem)の存在下でインキュベートし、37℃で60minインキュベートした後、50℃で60min、0.25%トリプシン(Gibco)で消化した。最後に、37℃で30min、DNaseI(50U/ml;Promega)で処理した後、次いで95℃で20min変性させた。線状化されたプラスミドpDC67/+SV40を用いて、標準曲線を作製した。プライマー−プローブセットは、サルウイルス40(SV40)ポリ(A)配列:rAAV−F:5’−AGCAATAGCATCACAAATTTCACAA−3’(配列番号4);rAAV−R:5’−GCAGACATGATAAGATACATTGATGAGTT−3’(配列番号5);rAAV−プローブ:5’6−FAM−AGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTC−TAMRA−3’(配列番号6)に特異的であった。PCR産物の増幅を、以下のサイクリングパラメータ:50℃で2minの1サイクル、95℃で10minの1サイクル;95℃で15secおよび60℃で60secの40サイクルを用いて達成した。
集密な293細胞(75cm2フラスコ中の2.4x106個の細胞)を、およそ20hにわたってDMEM+10%FCS中で培養した後、1プラーク形成単位/細胞(PFU/細胞)のHSV−1 MacIntyre株を90min感染させた。20μM(最終)濃度のATAを感染の90min後に添加した。感染細胞を感染の24h後に収穫した。十分な品質および量の全RNAを、製造業者の説明書に従ってRNeasy Plus Mini Kit(QIAGEN、Valencia、CA)を用いて懸濁液から単離した。全ゲノム発現プロファイリングを、Asuragen,Inc.によるAffimetrix Human U133 Plus 2.0Array上で行った。3μgの全RNAを入力材料として提供した。
6x105個の293細胞(AGTCから)を、50μMのATAを含むか、または含まないDMEMおよび10%FBSの存在下で1〜2時間、MOI 1でwtHSV−1 McIntyre株に感染させ、DMEMおよび10%FBSで40%に希釈した(最終ATA濃度20μM)。Qiagen RNeasyミニキットを用いて24h後に3回、全RNA試料を収穫し、カラム上でDNase処理し、溶出させた。溶出液RNAの3回試料を合わせ、分光光度計O.D.の読取りを260および280nmで溶出液に対して行い、濃度を決定した。試料RNAを、SABiosciences RT2第一鎖キットを用いて鋳型cDNAに変換した。次いで、cDNAを、ヒトJAK/STATシグナリング経路PCRアレイ(PAHS−039A)中で用いた。
Dulbecco’s Modified Eagles Medium(DMEM)(Hyclone、Waltham、MA)中のV27細胞を、1のMOIでHSV−1 d27.1ベクターに感染させ、10%ウシ胎仔血清(FBS)を用いて、または用いずに30μMのATA(Sigma、St.Louis、MO)で処理した。ATAを用いない対照実験も行った。V27細胞溶解物を感染の24時間後(h.p.i.)に得て、溶解物に対してウェスタンブロットを実行し、iNOSタンパク質を精製ウサギ抗iNOS/NOS TypeIIpAb(BD Biosciences、カタログ番号610332)を用いることにより検出した。結果を図1に示す。
ATAがV27細胞中でのrHSV−1 d27−1(d27−1)ベクター収量を増加させることができるかどうかを試験するために、ATAを感染(工程1)または希釈工程(工程2)の間に培地に印加した(図2A)。MOI=0.15でのd27−1感染を、最終培地容量の2/5で行い、残りの3/5の培地を希釈工程の間に添加した。
さらなる実験を行って、最適な条件およびHSV力価に対するFBSの存在または非存在の効果を決定した。ATA含有培地中の血清、特に、10%ウシ胎仔血清(FBS)の存在が、ATAにより誘導されるHSV−1力価収量のための最も重要なパラメータであった。希釈工程中の無血清培地中へのATAの追加(工程2でのATA)は、ウイルス力価の低下をもたらし、「DRP」力価は対照レベルより下まで低下し、PFU/ml力価は検出以下であった(図3)。無血清培地中のさらにより劇的な効果は、3μM濃度で出発するATAを感染工程1の間に追加した場合に認められ(データは示さない)、DRP/mlおよびPFU/ml力価の両方が検出限界以下であった。
図4Aは、wtHSV−1増殖を許容する細胞系:293細胞、HeLa細胞およびVero細胞におけるwtHSV−1株、KOSおよびMcIntyreの両方の上清力価を示す。wtHSV−1株KOSおよびMcIntyreを、50μMのATAを工程1で添加し、工程2の後に最終濃度を20μMのATAに希釈するATA−HSVプロトコールを用いて、293細胞、HeLa細胞およびVero細胞中で増殖させた。ウイルスを含有する上清を、上記のように3日後に収穫した。ATAは、Vero細胞においては両方のウイルス型の収量を増加させた。wtHSV−1 McIntyre株は293細胞中ではATA誘導後に最も高い力価に達したが、ATAは293細胞中ではHSV−1 KOS成長を阻害すると考えられた。最後に、ATAはまた、HeLa細胞中では両方の型のHSV−1ウイルスを阻害すると考えられた。二元配置ANOVA;Bonferroni検定;−ATA対+ATA:***p<0.001、ns:p>0.05;n=4独立実験。
HSV産生中のATAの存在がHSVベクターを用いて産生されたrAAVビリオンの収量に影響したかどうかを決定するために、rAAV力価が、293細胞中で産生されたrHSV−1ストックから存在するATA残留物により影響されたことを示す以下の実験を実施した(図5A)。異なる濃度のATA濃度(12μMまたは20μM最終濃度)の下で製造されたATAを含有するrHSV−1ストックを用いて60mmプレート中の293細胞中でrHSV−rep2/cap2およびrHSV−EGFPベクターの同時感染により、rAAV−GFPベクターを産生した(材料および方法を参照されたい)。rAAV DRP/ml力価は、感染中に20μM ATAを添加して製造された(ATA工程1)rHSV−rep2/cap2ストックを用いた場合に、rAAV産生中のATA濃度がおよそ3μMであった「ATAなし」対照(*p<0.05)に対してわずかに1.3倍増加した。有意なrAAV力価の増加は、12μM ATAを用いて製造されたrHSV−rep2/cap2ストックを用いた場合に検出された(図5A)。一元配置ANOVA;Tukey検定:*p<0.05;20μM ATA対ATAなし;n=4独立実験。
ヒトゲノムアレイによるATAの作用機序ならびにVeroとV27がサル由来細胞系であるという事実を明らかにするために、wtHSV−1増殖を、いくつかのヒト細胞系中で試験した(図4A〜4B)。2つの野生型HSV−1(wtHSV−1)株、KOSおよびMcIntyreの増殖を、3つの細胞系:ヒト胚性腎293(293細胞)、ヒト子宮頸がんHeLa細胞およびアフリカミドリザル腎上皮Vero細胞において比較した。実験を、50μMのATAを感染工程中に追加し、20μM ATA濃度にさらに希釈するATA Iプロトコールスキームに従って10%FBS含有培地中で実行した(材料および方法を参照されたい)。ヒト由来293細胞においては、wtHSV−1 McIntyreウイルス力価のみが、1.4x108DRP/mlから1.2x109DRP/mlまで、ATAにより有意に増加した(***p<0.001)。Vero細胞においては、ATAはwtHSV−1 KOS株においてのみ、1.7x108DRP/mlから9.1x108DRP/mlまで力価を有意に増加させた(***p<0.001;両統計値:二元配置ANOVA、Bonferroni検定;n=4)。驚くべきことに、HeLa細胞においては、ATAはKOSとMcIntyre HSV−1株の両方の増殖を阻害すると考えられた。
iNOS阻害剤であるデキサメタゾンもまた培養物中でのrHSV収量を増加させたかどうかを決定するために、以下の実験を行った。図6は、d27−1/GFP HSV−1ウイルス力価に対するデキサメタゾンの効果を示す。最終的な力価d27−1/GFP HSV−1は、一般に、未処理対照と比較してdex予備処理または処理の後にわずかに上昇した。
iNOS阻害剤であるバルプロ酸もまた培養物中でのrHSV収量を増加させたかどうかを決定するために、以下の実験を行った。図7は、d27−1/GFP HSV−1ウイルス力価に対するバルプロ酸(VA)による予備処理の効果を示す。5mM濃度のVAは、d27−1/GFP HSV−1の力価をわずかに上昇させたが、5mM未満およびそれを超える濃度は、未処理の対照と比較した場合、d27−1/GFP HSV−1力価に対する阻害効果を有すると考えられた。
Claims (14)
- アウリントリカルボン酸を含む細胞培養物中でウイルスを培養することを含む、ウイルスを産生するための方法。
- 前記ウイルスがヘルペスウイルスである、請求項1に記載の方法。
- 前記ヘルペスウイルスが単純ヘルペスウイルス1型(HSV−1)である、請求項2に記載の方法。
- 前記HSV−1が野生型HSV−1である、請求項3に記載の方法。
- 前記HSV−1が組換えHSV−1ベクターである、請求項3に記載の方法。
- 前記組換えHSV−1ベクターがHSV−1 d27.1ベクターである、請求項5に記載の方法。
- 前記ウイルスが293、HeLaまたはVero細胞中で培養される、請求項1〜6のいずれか1項に記載の方法。
- 前記ウイルスがV27細胞中で培養される、請求項7に記載の方法。
- (a)V27細胞をHSV−1 d27.1ベクターに感染させること;および
(b)前記感染したV27細胞を、アウリントリカルボン酸、バルプロ酸またはデキサメタゾンを含む細胞培養物中で培養すること
を含む、HSV−1 d27.1ベクターを培養するための方法。 - 細胞培養物がアウリントリカルボン酸を含む、請求項9に記載の方法。
- 前記細胞培養物が血清をさらに含む、請求項9または10に記載の方法。
- 血清がウシ胎仔血清である、請求項11に記載の方法。
- アウリントリカルボン酸および293、HeLaまたはVero細胞を含む細胞培養物。
- V27細胞を含む、請求項13に記載の細胞培養物。
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Publication number | Priority date | Publication date | Assignee | Title |
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CA2966620A1 (en) | 2014-11-05 | 2016-05-12 | Voyager Therapeutics, Inc. | Aadc polynucleotides for the treatment of parkinson's disease |
SG11201703419UA (en) | 2014-11-14 | 2017-05-30 | Voyager Therapeutics Inc | Modulatory polynucleotides |
US10597660B2 (en) | 2014-11-14 | 2020-03-24 | Voyager Therapeutics, Inc. | Compositions and methods of treating amyotrophic lateral sclerosis (ALS) |
US11697825B2 (en) | 2014-12-12 | 2023-07-11 | Voyager Therapeutics, Inc. | Compositions and methods for the production of scAAV |
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Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH08502043A (ja) * | 1992-07-31 | 1996-03-05 | プレジデント アンド フェローズ オブ ハーバード カレッジ | ヘルペスウイルスワクチン |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5139941A (en) | 1985-10-31 | 1992-08-18 | University Of Florida Research Foundation, Inc. | AAV transduction vectors |
WO1992001070A1 (en) | 1990-07-09 | 1992-01-23 | The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce | High efficiency packaging of mutant adeno-associated virus using amber suppressions |
US5173414A (en) | 1990-10-30 | 1992-12-22 | Applied Immune Sciences, Inc. | Production of recombinant adeno-associated virus vectors |
RU2012335C1 (ru) | 1991-07-04 | 1994-05-15 | Уфимский научно-исследовательский институт глазных болезней | Ингибитор вируса простого герпеса |
DE69233013T2 (de) | 1991-08-20 | 2004-03-04 | The Government Of The United States Of America As Represented By The Secretary Of National Institute Of Health, Office Of Technology Transfer | Adenovirus vermittelter gentransfer in den gastrointestinaltrakt |
US7223411B1 (en) | 1992-07-31 | 2007-05-29 | Dana-Farber Cancer Institute | Herpesvirus replication defective mutants |
US6833271B2 (en) | 1996-12-04 | 2004-12-21 | Medi-Cult A/S | Serum-free cell culture media |
GB9816856D0 (en) | 1998-08-03 | 1998-09-30 | Univ London | Cell lines for virus growth |
US6783972B1 (en) * | 1998-09-22 | 2004-08-31 | University Of Florida Research Foundation | Methods for large-scale production of recombinant AAV vectors |
US6593123B1 (en) | 2000-08-07 | 2003-07-15 | Avigen, Inc. | Large-scale recombinant adeno-associated virus (rAAV) production and purification |
US7091029B2 (en) * | 2002-09-23 | 2006-08-15 | Applied Genetics Technologies Corporation | High titer recombinant AAV production |
US7427396B2 (en) | 2004-06-03 | 2008-09-23 | Genzyme Corporation | AAV vectors for gene delivery to the lung |
CN100348718C (zh) * | 2005-10-18 | 2007-11-14 | 中国人民解放军军事医学科学院生物工程研究所 | 一种支持hek293细胞贴附培养的无动物来源成分无血清培养基 |
CA2692460A1 (en) * | 2007-06-29 | 2009-01-08 | Korea Research Institute Of Chemical Technology | Hiv reverse transcriptase inhibitors |
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Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH08502043A (ja) * | 1992-07-31 | 1996-03-05 | プレジデント アンド フェローズ オブ ハーバード カレッジ | ヘルペスウイルスワクチン |
Non-Patent Citations (6)
Title |
---|
CHEN, C., ET AL.: "Inhibition of cytokine-induced JAK-STAT signalling pathways by an endonuclease inhibitor aurintricar", BRITISH JOURNAL OF PHARMACOLOGY, vol. 137, JPN6017039979, 2002, pages 1011-1020 * |
ERLANDSSON, AC, ET AL.: "Herpes simplex virus 1 infection and glucocorticoid treatment regulate viral yield, glucocorticoid r", JOURNAL OF ENDOCRINOLOGY, vol. 175, JPN6017039974, 2002, pages 165-176 * |
KODUKULA, P., ET AL.: "Macrophage Control of Herpes Simplex Virus Type 1 Replication in the Peripheral Nervous System", THE JOURNAL OF IMMUNOLOGY, vol. 162, JPN6017039977, 1999, pages 2895-2905 * |
MYSKIW CHAD, JOURNAL OF VIROLOGY, vol. V81 N6, JPN5016003187, March 2007 (2007-03-01), pages 027 - 3032 * |
NEYTS, J., ET AL.: "Poly(hydroxy)carboxylates as selective inhibitors of cytomegalovirus and herpes simplex virus replic", ANTIVIRAL CHEMISTRY & CHEMOTHERAPY, vol. 3(4), JPN6017039975, 1992, pages 215-222 * |
OTSUKI, A., ET AL.: "Histone Deacetylase Inhibitors Augment Antitumer Efficacy of Herpes-based Oncolytic Viruses", MOLECULAR THERAPY, vol. 16(9), JPN6017039972, 2008, pages 1546-1555 * |
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