JP5833145B2 - 免疫原性bordetellabronchiseptica組成物 - Google Patents
免疫原性bordetellabronchiseptica組成物 Download PDFInfo
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- JP5833145B2 JP5833145B2 JP2013552317A JP2013552317A JP5833145B2 JP 5833145 B2 JP5833145 B2 JP 5833145B2 JP 2013552317 A JP2013552317 A JP 2013552317A JP 2013552317 A JP2013552317 A JP 2013552317A JP 5833145 B2 JP5833145 B2 JP 5833145B2
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Description
一つの態様において、本発明は、特に、Bordetella bronchiseptica及び単離されたパータクチン抗原を含んでなる免疫原性組成物を提供する。もう一つの態様において、前記パータクチン抗原は、組換えタンパク質である。もう一つの態様において、前記パータクチン抗原は、Bordetella bronchisepticaからのものである。もう一つの態様において、前記パータクチン抗原は、p68である。もう一つの態様において、前記組成物は、単離されたBsp22抗原を更に含んでなる。
“治療的有効量”は、本明細書中で使用する場合、抗原又はワクチン或いは組成物を投与される対象者(例えばイヌ)に免疫反応を誘発するものである抗原又はワクチン或いは組成物の、ウイルス、細菌、寄生虫又は真菌のような病原体による感染によって起こされる不都合な健康上の影響又はその合併症を含む疾病の徴候又は症状を予防又は寛解するために十分である量を指す。体液性免疫又は細胞仲介免疫、或いは体液性免疫及び細胞仲介免疫の両方を誘発することができる。抗原、ワクチン、又は組成物に対する動物の免疫原性反応は、抗体力価の測定、リンパ球増殖アッセイによって間接的に、又は野生型株を用いたチャレンジ後の徴候及び症状をモニターすることにより直接的に評価することができる。ワクチン又は組成物によって与えられる保護免疫は、チャレンジ生物の排出の減少、並びに/又は死亡率、罹患率、体温、及び対象者の全体的身体状態、健康、及び能力のような臨床的徴候の減少を測定することによって評価することができる。治療的に有効であるワクチン又は組成物の量は、使用される特定の免疫原、又は対象者の状態によって変更することができ、そして当業者によって決定されることができる。
本開示は、Bordetella bronchiseptica調製物と組合せた単離されたパータクチン抗原の封入が実質的に改善された効力及び安全性をもたらすという予期しない発見に基づく。即ち、細菌のチャレンジに先立ちイヌに非経口投与した場合、組成物は、気管気管支炎の発症を予防し、そして不都合な副作用をもたらさない。
本発明によって包含される免疫原性組成物及びワクチンは、CIRDCに対する有効な免疫反応を誘発するために動物に投与することができる。従って、本発明は、治療的有効量の本明細書中に記載される免疫原性組成物又はワクチンを動物に投与することによる、有効な免疫反応を刺激する方法を提供する。
動物において誘発された免疫反応の程度及び特質は、各種の技術を使用することによって評価することができる。例えば、血清を、接種された動物から収集し、そして免疫原に対して特異的な抗体の存在又は非存在を試験することができる。細胞性免疫反応の誘発を示すリンパ系組織中の反応性細胞障害性Tリンパ球(CTL)の検出は、T細胞増殖のようなアッセイによって達成することができる。関連する技術は、当技術分野において十分に記載されている。
免疫原性組成物又はワクチンを、更なる組成物又は化合物と組合せて−例えば、特定の疾病又は状態を治療する目的のために−投与することが好ましいことであり得る限り、免疫原性組成物又はワクチンを、都合よく、組成物の投与又は同時投与のために適したキットの形態に含めるか、又は組合せることができることは本発明の範囲内である。
抗体は、モノクローナル、ポリクローナル、又は組換え型のいずれかであることができる。抗体は、免疫原又はその一部に対して調製することができる。例えば、免疫原のアミノ酸配列に基づいて、又はクローニング技術によって組換え的に調製された合成ペプチド、或いは天然の遺伝子産物及び/又はその一部を、単離し、そして免疫原として使用することができる。免疫原は、当業者にとって周知の標準的な抗体産生技術によって、抗体を産生するために使用することができる。抗体の断片も、当業者にとって既知の方法によって抗体から調製することができ、そしてFab、F(ab’)2、及びFv断片を含む。
生後概略8週間の、そして良好な一般的健康状態の40匹のイヌを、Bordetella bronchisepticaについて微小凝集試験(MAT)によって、そしてイヌ呼吸器コロナウイルス(CRCoV)について間接蛍光抗体アッセイ(IFA)によって予備スクリーニングした。血清中和(SN)を、更に抗体レベルを評価するために使用した。0日目、全てのイヌは、MAT(≦16)によって決定されるようにBordetella bronchisepticaに対する抗体について陰性であり、そしてIFA(<40)によって決定されるようにCRCoVに対する抗体について陰性であった。全てのイヌは、更に一回目のワクチン接種(0日目)に先立つ鼻腔拭取り検体の単離試験によって決定されるように、Bordetella bronchiseptica及びCRCoVを持っていなかった。
T02及びT03群の試験ワクチンは、二回目の投与後、全て(100%)のワクチン接種したイヌに抗体反応を誘発し、ウイルス抗原に対する活性な免疫化を示した。抗体反応は、大多数のワクチン接種したイヌにおいて二回目のワクチン接種後増加し、二回目のワクチン接種の追加免疫効果を示した。ウイルス画分の中での抗体反応が、多数のウイルス及び細菌(B.bronchiseptica)抗原の存在下で達成され、免疫学的干渉の欠如を示したことを注記することは重要である。MATの血清学は、Bordetellaに対する保護に相関しないが、しかしむしろ適した研究動物を登録するための価値あるスクリーニングの道具である。結論として、ワクチン接種したイヌにおける免疫学的反応に基づけば、ウイルス抗原の効力は、5種多価ワクチンにおいて予測される。
A.一価:
この研究の目的は、病原性B.bronchisepticaチャレンジに対する生後8週間のイヌにおける、Bordetella bronchiseptica抽出物−サブユニットワクチンの免疫の期間を証明することにある。
この研究の目的は、イヌにおける多価呼吸器混合ワクチンの免疫の期間を証明することである。ワクチンは、パータクチン、Bsp22、又は両方のいずれかの組換え抗原で補充された次の抗原性成分を含有する:改変生CAV−2、改変生CPIV、不活化CIV、不活化CRCoV及びBordetella bronchiseptica抽出物。
Claims (18)
- Bordetella bronchiseptica及び単離されたパータクチン抗原を含んでなる、免疫原性組成物。
- 前記パータクチン抗原が、Bordetella bronchisepticaタンパク質である、請求項1に記載の免疫原性組成物。
- 前記パータクチン抗原が、組換えp68タンパク質である、請求項2に記載の免疫原性組成物。
- 前記Bordetella bronchisepticaが、バクテリン又は細菌抽出物である、請求項1ないし3のいずれか1項に記載の免疫原性組成物。
- 前記組成物が、アジュバント非添加である、請求項1ないし4のいずれか1項に記載の免疫原性組成物。
- 前記パータクチン抗原が、1μgないし30μgの間で存在する、請求項1ないし5のいずれか1項に記載の免疫原性組成物。
- 前記パータクチン抗原が、尿素でパータクチン封入体を可溶化することによって調製され、該組成物がパータクチン抗原の凝集物を含まない、請求項1ないし6のいずれか1項に記載の免疫原性組成物。
- 該パータクチン抗原が、脂質付加されている、請求項1ないし7のいずれか1項に記載の免疫原性組成物。
- 前記組成物が、イヌへの非経口投与のための希釈剤又は賦形剤を更に含んでなる、請求項1ないし8のいずれか1項に記載の免疫原性組成物。
- 前記組成物が、脂質付加された単離されたBsp22抗原を更に含んでなる、請求項1ないし9のいずれか1項に記載の免疫原性組成物。
- 前記組成物が、イヌパラインフルエンザウイルス(CPIV)、イヌアデノウイルス−2(CAV−2)、イヌ呼吸器コロナウイルス(CRCoV)、Mycoplasma cynos(M.Cynos)及びイヌインフルエンザウイルス(CIV)からなる群から選択されるイヌ呼吸器病原体からの抗原を更に含んでなる、請求項1ないし10のいずれか1項に記載の免疫原性組成物。
- 前記組成物が、イヌ呼吸器病原体からの抗原の少なくとも二つ、三つ又は四つを含んでなる、請求項11に記載の免疫原性組成物。
- 前記組成物が、イヌパラインフルエンザウイルス(CPIV)、イヌアデノウイルス−2(CAV−2)、イヌ呼吸器コロナウイルス(CRCoV)及びイヌインフルエンザウイルス(CIV)からの抗原を含んでなる、請求項11に記載の免疫原性組成物。
- イヌにおけるイヌ呼吸器病原体による感染の治療又は予防のための、請求項1ないし13のいずれか1項に記載の免疫原性組成物の使用。
- 前記組成物が、前記イヌにおける前記イヌ呼吸器病原体による感染を、6ヶ月又はそれより多い期間予防する、請求項14に記載の使用。
- 該イヌ呼吸器病原体が、Bordetella bronchisepticaである、請求項14又は15に記載の使用。
- 該イヌ呼吸器病原体が、イヌパラインフルエンザウイルス(CPIV)、イヌアデノウイルス−2(CAV−2)、イヌ呼吸器コロナウイルス(CRCoV)及びイヌインフルエンザウイルス(CIV)の少なくとも一つ、二つ、三つ又は四つを更に含んでなる、請求項16に記載の使用。
- 該組成物が、複数の前記病原体による感染を治療又は予防する、イヌ感染性呼吸器病症候群(CIRDC)の治療又は予防のための、請求項14−17のいずれか1項に記載の使用。
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