JP5500762B2 - エシェリキア・コリおよびサルモネラに対する鳥用混合ワクチン - Google Patents
エシェリキア・コリおよびサルモネラに対する鳥用混合ワクチン Download PDFInfo
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- JP5500762B2 JP5500762B2 JP2006532983A JP2006532983A JP5500762B2 JP 5500762 B2 JP5500762 B2 JP 5500762B2 JP 2006532983 A JP2006532983 A JP 2006532983A JP 2006532983 A JP2006532983 A JP 2006532983A JP 5500762 B2 JP5500762 B2 JP 5500762B2
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Description
本発明はサルモネラ菌およびエシェリキア・コリ感染症に対する鳥用混合ワクチンに関する。特に、本発明は、サルモネラ菌およびエシェリキア・コリの変異型生、弱毒化株をそれぞれ含む家禽用ワクチン組成物に関する。本発明はまた、サルモネラ菌およびエシェリキア・コリの弱毒化微生物を含むワクチンを投与することによって、ニワトリを含む家禽における細菌性疾患を予防する方法に関する。
養鶏業において、孵化したばかりのヒナはサルモネラ菌に特に感染しやすい。この細菌は糞便物から広がるため、幼若動物では土壌から、また場合によっては汚染された加工飼料から感染することがあり、高い死亡率とそれに伴う深刻な経済的影響をもたらし得る。サルモネラ菌に加え、家禽舎の環境では糞便汚染を通じて多数のエシェリキア・コリ(E. Coli)を維持するため、呼吸器管および腸管を介して家禽の全身性感染を引き起こす。エシェリキア・コリ感染症はエシェリキア・コリ症と称される。その結果生じる菌血症は敗血症に進行し死に至るか、感染症が漿膜表面、心膜、関節および他の臓器に及ぶ。このようにエシェリキア・コリおよびサルモネラ菌の疾患は、養鶏業にとって深刻な脅威として継続して存在する。
本発明の利点は、生ワクチン組成物が宿主中で細胞性および体液性免疫応答の両方を提供することである。
本発明は、安全かつ有効な鳥用ワクチン組成物であって、免疫原的有効量のエシェリキア・コリ遺伝子欠失変異微生物およびサルモネラ・ティフィムリウム遺伝子欠失変異微生物の組合せ、並びに医薬上許容される担体を含む組成物を提供する。
国内家禽における鳥類エシェリキア・コリ症は、血清型O78、O1、O2および特に未分類株を含む強毒のエシェリキア・コリとしばしば関係している。感染症は、しばしば別の家禽の集団疾患(community diseases)への暴露、または集団疾患による感染に続いて、通常、呼吸器管を通じて発症する。ニワトリでは、エシェリキア・コリ症は一般に3〜10週齢のブロイラーに罹患し、高い罹病率および死亡率を伴う。鳥類エシェリキア・コリ症の最も重篤な症状は、心膜炎、肝周囲炎および気嚢炎により特徴付けられる敗血症である。他の問題としては関節炎および蜂巣炎がある。家禽からのエシェリキア・コリ単離体は、例えばアンピリシン、クロラムフェニコール、オキシテトラサイクリン、ネオマイシン、ゲンタマイシン、ニトロフラン、ナリジキシ酸、ポリミキシンB、スルホンアミドまたはその類似物などの薬剤に対してしばしば抵抗性である。しかしながら、2003年3月27日にアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託され、ATCC番号PTA−5094を有する株の識別特徴(identifying characteristics)を有する、エシェリキア・コリaroA遺伝子欠失変異微生物、エシェリキア・コリaroA−は、ニワトリを含む家禽における鳥類エシェリキア・コリ症に対するワクチンとしての使用に安全かつ有効である。エシェリキア・コリaroA− PTA−5094ワクチン免疫原は、それをニワトリに投与した場合に、良い細胞性および体液性免疫反応を提供する。さらに、かかるワクチンは容易に製造でき、集団適用、即ち噴霧または飲水を介した投与が可能である。エシェリキア・コリaroA− PTA−5094ワクチンおよびその構築は、本出願と同時に出願した同時係属の特許出願番号______にも記載されており、出展明示により本発明の一部とする。加えて、本発明はさらに、本明細書中に記載した混合ワクチンの一部の免疫原として、欠失変異体、特にaroA欠失変異体を含む他の生、弱毒化エシェリキア・コリ遺伝子変異体が考えられる。
aroA遺伝子欠失エシェリキア・コリ変異体の構築
I)受容菌
親生物は、獣医学研究所(VLA)、アドレストン(Addlestone)、サリー州、英国に寄託され、1995年にVLAで血清型を決定された鳥類エシェリキア・コリ症の臨床例から単離されたエシェリキア・コリの鳥類単離体である。親株は、1日齢SPFヒナにおけるそのコロニー形成性、侵入性、持続性および病原性について、並びにその抗生物質感受性パターンについてのイン・ビトロ(in vitro)特徴づけにより、選択された。受容菌株は、形質転換供与菌(100bp欠失したaroAを有するPNG101を含むエシェリキア・コリK12 S17 λ pir)と野生型親株(野生型エシェリキア・コリ単離体EC34195)との間の接合により作製された。
通常の芳香族生合成経路の酵素である3−ホスホエノールピルビルシキメート−5−リン酸合成酵素をコードするaroA遺伝子は、serC−aroAオペロン中のSerCに隣接し且つプロモーターの末端に位置する。受容菌におけるaroA遺伝子機能の喪失により、チロシン、フェニルアラニン、トリプトファン、p−アミノ安息香酸(PABA)および2,3−ジヒドロキシ安息香酸を含む芳香族代謝物の要求性となる。脊椎動物の組織中で見られない代謝物であるPABAの要求性は、インビボ(in vivo)増殖を弱めることとなる。
a)PCRプライマーをSrfIおよびBglII制限部位並びに終止コドンが組み込まれるように設計して、前記の単離された家禽エシェリキア・コリO78からaroA遺伝子の5’端および3’端の約650bpの2個別々のPCR産物を増幅する。
自己不活化ベクター(SacB、PNG101)は中間のクローニングベクターであった。供与菌(100bp欠失したaroAを有するPNG101を含むS17)と野生型エシェリキア・コリ単離体との間で接合を行った。
原株(master seed)の調製
エシェリキア・コリaroA−株(実施例1にて構築したもの)をトリプシン大豆寒天プレート上で一度生育させ、トリプシン大豆肉汁培地中にて3回継代する。培養物をガラス瓶中に分け、密閉して凍結乾燥させる。
ニワトリにおけるエシェリキア・コリおよびサルモネラ菌感染症に対するエシェリキア・コリ遺伝子欠失変異微生物およびサルモネラ・ティフィムリウム遺伝子欠失変異微生物の組合せの有効性の評価
本評価において、使用するエシェリキア・コリ遺伝子欠失変異微生物は、上記実施例1および2にて作製されたエシェリキア・コリaroA−株であり、使用するサルモネラ・ティフィムリウム遺伝子欠失変異微生物は、米国特許第6,231,871号B1に記載のように調製されたサルモネラ・ティフィムリウムSTM−1である。
Claims (9)
- 免疫原的有効量の受入番号PTA−5094でアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託された微生物および受入番号N93/43266でオーストラリアン・ガバメント・アナリティカル・ラボラトリーズ(AGAL)に寄託されたサルモネラ・ティフィムリウムSTM−1の組合せならびに医薬上許容される担体を含む、鳥用ワクチン組成物。
- 担体が組織または細胞培地での使用に好適な平衡塩類溶液である、請求項1記載の組成物。
- 担体が蒸留水である、請求項1記載の組成物。
- 免疫原的有効量が5.0×102cfu〜5.0×1010cfuの受入番号PTA−5094でアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託された微生物および5.0×102cfu〜5.0×1010cfuの受入番号N93/43266でオーストラリアン・ガバメント・アナリティカル・ラボラトリーズ(AGAL)に寄託されたサルモネラ・ティフィムリウムSTM−1を提供するのに十分な量である、請求項1〜3のいずれか1項に記載の組成物。
- 免疫原的有効量が3.0×106〜6.0×106cfuの受入番号PTA−5094でアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託された微生物および2.0×106〜6.0×106cfuの受入番号N93/43266でオーストラリアン・ガバメント・アナリティカル・ラボラトリーズ(AGAL)に寄託されたサルモネラ・ティフィムリウムSTM−1を提供するのに十分な量である、請求項1〜3のいずれか1項に記載の組成物。
- エシェリキア・コリおよびサルモネラ・ティフィムリウムによってもたらされる疾患または感染症に対して家禽を保護する方法であって、免疫原的有効量の受入番号PTA−5094でアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託された微生物および受入番号N93/43266でオーストラリアン・ガバメント・アナリティカル・ラボラトリーズ(AGAL)に寄託されたサルモネラ・ティフィムリウムSTM−1の組合せを含む鳥用ワクチン組成物を該家禽に投与することを含む、方法。
- 家禽が、ニワトリ、アヒル、ガチョウ、シチメンチョウ、バンタム、ウズラ、キジおよびハトからなる群から選択される、請求項6記載の方法。
- 該鳥用ワクチン組成物が集団適用にて投与される、請求項6または7記載の方法。
- 免疫原的有効量が3.0×106〜6.0×106cfuの受入番号PTA−5094でアメリカン・タイプ・カルチャー・コレクション(ATCC)に寄託された微生物および2.0×106〜6.0×106cfuの受入番号N93/43266でオーストラリアン・ガバメント・アナリティカル・ラボラトリーズ(AGAL)に寄託されたサルモネラ・ティフィムリウムSTM−1を提供するのに十分な量である、請求項6〜8のいずれか1項に記載の方法。
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US20080260777A1 (en) * | 2007-04-19 | 2008-10-23 | Sotomayor Konky | Composition and method for controlling intestinal pathogenic organisms |
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JP2012131796A (ja) | 2012-07-12 |
BRPI0410355B1 (pt) | 2016-09-13 |
JP2006528979A (ja) | 2006-12-28 |
AU2004240585B2 (en) | 2010-08-19 |
AR044339A1 (es) | 2005-09-07 |
TW200503755A (en) | 2005-02-01 |
CN1787836A (zh) | 2006-06-14 |
US20040234550A1 (en) | 2004-11-25 |
CN102274495A (zh) | 2011-12-14 |
AU2004240585A1 (en) | 2004-12-02 |
CA2524939C (en) | 2012-12-04 |
WO2004103401A3 (en) | 2005-02-10 |
BRPI0410355A (pt) | 2006-07-04 |
ZA200509982B (en) | 2011-05-25 |
WO2004103401A2 (en) | 2004-12-02 |
CA2524939A1 (en) | 2004-12-02 |
NZ543558A (en) | 2008-02-29 |
EP1622643A2 (en) | 2006-02-08 |
KR101150318B1 (ko) | 2012-06-08 |
US7297338B2 (en) | 2007-11-20 |
KR20060012001A (ko) | 2006-02-06 |
MXPA05012219A (es) | 2006-02-08 |
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