TWI456057B - 具有增強之逃脫核內體能力的重組卡介苗（ｂｃｇ）菌株 - Google Patents

具有增強之逃脫核內體能力的重組卡介苗（ｂｃｇ）菌株 Download PDF

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TWI456057B
TWI456057B TW094141904A TW94141904A TWI456057B TW I456057 B TWI456057 B TW I456057B TW 094141904 A TW094141904 A TW 094141904A TW 94141904 A TW94141904 A TW 94141904A TW I456057 B TWI456057 B TW I456057B
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functional
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nucleic acid
protein
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Rong Gai Sun
David Michael Hone
Jerald C Sadoff
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Aeras Global Tb Vaccine Found
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Claims (34)

  1. 一種分枝桿菌，它經基因工程改造成包括編碼一可表現及可分泌之功能性核內體溶解性(endosomalytic)蛋白質的核酸序列，此蛋白質在pH6-8處呈活性，且該可表現及可分泌之功能性核內體溶解性蛋白質為穿破溶解素(perfringolysin)或其一種功能性變異體，其中，所指之可表現及可分泌之功能性核內體溶解性穿破溶解素之一種功能性變異體係得自對穿破溶解素的胺基酸序列進行取代、刪除或加入而且該穿破溶解素之一種功能性變異體顯示至少80%對穿破溶解素的胺基酸序列之同源性。
  2. 如申請專利範圍第1項之分枝桿菌，其中，所指之可表現及可分泌之功能性核內體溶解性蛋白質的一胺基酸序列乃如SEQ ID NO：2所示者。
  3. 如申請專利範圍第1項之分枝桿菌，其中，所指之核酸序列專一性地針對穿破溶解素或其一變異體。
  4. 如申請專利範圍第3項之分枝桿菌，其中，所指之核酸序列係選自SEQ ID NO：1及SEQ ID NO：3所組成之群體。
  5. 如申請專利範圍第1項之分枝桿菌，其中，所指之分枝桿菌係經基因工程改造成包括編碼一細胞凋亡性蛋白質或細胞凋亡之一功能性增強物的核酸序列。
  6. 如申請專利範圍第5項之分枝桿菌，其中，所指之細胞凋亡性蛋白質或所指之細胞凋亡之功能性增強物係選自caspase 8，死亡受體-5，Fas及Fas細胞質功能域/CD4外 功能域融合蛋白質(Fas cytoplasmic domain/CD4 ectodomain fusion protein)。
  7. 如申請專利範圍第1項之分枝桿菌，其中，所指之分枝桿菌係經基因工程改造成包括編碼一或更多所關注之蛋白質的核酸序列。
  8. 如申請專利範圍第7項之分枝桿菌，其中，所指之一或更多所關注之蛋白質包括結核桿菌(Mtb)抗原。
  9. 如申請專利範圍第1項之分枝桿菌，其中，所指之分枝桿菌係經基因工程改造成以包括編碼一種細胞凋亡性蛋白質或細胞凋亡之一種功能性增強物的核酸序列；以及編碼一或更多所關注之蛋白質的核酸序列。
  10. 如申請專利範圍第9項之分枝桿菌，其中，所指之編碼一或更多所關注之蛋白質的核酸序列包括編碼抗原、免疫調節因子或佐劑之任何組合的至少一傳遞者核苷酸序列(passenger nucleotide sequence)。
  11. 如申請專利範圍第1項之分枝桿菌，其中，該可表現及可分泌之功能性核內體溶解性蛋白質在被所指之分枝桿菌屬感染之細胞的核內體內呈現之pH值6-8處呈活性。
  12. 如申請專利範圍第1項之分枝桿菌，其中，所指之分枝桿菌係BCG。
  13. 一種使一分枝桿菌能夠由核內體脫逃之方法，它包含下列之步驟： 以基因工程改造所指之分枝桿菌，使之含有，編碼一可表現及可分泌之功能性核內體溶解性蛋白質的核酸序列，且該功能性核內體溶解性蛋白質係穿破溶解素O(perfringolysin O)。
  14. 如申請專利範圍第13項之方法，其中，所指之功能性核內體溶解性蛋白質係被SEQ ID NO：3編碼之一突變之穿破溶解素O。
  15. 如申請專利範圍第13項之方法，其中，所指之分枝桿菌係經減毒之分枝桿菌。
  16. 如申請專利範圍第15項之方法，其中，所指之經減毒之分枝桿菌係BCG。
  17. 如申請專利範圍第13項之方法，其中，所指之分枝桿菌係經基因工程改造成以包括編碼一細胞凋亡性蛋白質或細胞凋亡之一功能性增強物的核酸序列。
  18. 如申請專利範圍第17項之方法，其中，所指之細胞凋亡性蛋白質或細胞凋亡之功能性增強物係選自caspase 8，死亡-受體-5，Fas及Fas細胞質功能域/CD4外功能域融合蛋白質。
  19. 如申請專利範圍第13項之方法，其中，所指之分枝桿菌係經基因工程改造成包括編碼一或更多所關注之蛋白質的核酸序列。
  20. 如申請專利範圍第19項之方法，其中，所指之一或更多所關注之蛋白質包括結核桿菌(Mtb)抗原。
  21. 如申請專利範圍第13項之方法，其中，所指之分 枝桿菌係經基因工程改造成包括編碼一細胞凋亡性蛋白質或細胞凋亡之一功能性增強物的核酸序列；以及編碼一或更多所關注之蛋白質的核酸序列。
  22. 如申請專利範圍第21項之方法，其中，所指之編碼一或更多所關注之蛋白質的核酸序列包括編碼抗原、免疫調節因子或佐劑之任何組合的至少一傳遞者核苷酸序列。
  23. 一種組成物，它包含一種分枝桿菌，它經基因工程改造成包括編碼一可表現及可分泌之功能性核內體溶解性蛋白質的核酸序列，此蛋白質在pH6-8處呈活性，且該功能性核內體溶解性蛋白質係穿破溶解素O(perfringolysin O)。
  24. 如申請專利範圍第23項之組成物，其中，所指之功能性核內體溶解性蛋白質係被SEQ ID NO：3所編碼之一種突變穿破溶解素O。
  25. 如申請專利範圍第23項之組成物，其中，被所指之分枝桿菌表現之所指功能性核內體溶解性蛋白質的表現乃使得所指之重組分枝桿菌由核內體脫逃。
  26. 如申請專利範圍第23項之組成物，其中，所指之分枝桿菌係經減毒之分枝桿菌。
  27. 如申請專利範圍第26項之組成物，其中，所指之經減毒之分枝桿菌係BCG。
  28. 如申請專利範圍第23項之組成物，其中，所指之 分枝桿菌係經基因工程改造以包括編碼一細胞凋亡性蛋白質或細胞凋亡之一功能性增強物之核酸序列。
  29. 如申請專利範圍第28項之組成物，其中，所指之細胞凋亡性蛋白質或所指之細胞凋亡之功能性增強物係選自caspase 8，死亡一受體-5，Fas及Fas細胞質功能域/CD4外功能域融合蛋白質。
  30. 如申請專利範圍第23項之組成物，其中，所指之分枝桿菌係經基因工程改造成包括編碼一或更多所關注之蛋白質之核酸序列。
  31. 如申請專利範圍第30項之組成物，其中，所指之一或更多所關注之蛋白質包括結核桿菌(Mtb)抗原。
  32. 如申請專利範圍第23項之組成物，其中，所指之分枝桿菌係經基因工程改造成包括編碼一細胞凋亡性蛋白質或細胞凋亡之一功能性增強物的核酸序列；以及編碼一或更多所關注之蛋白質的核酸序列。
  33. 如申請專利範圍第32項之組成物，其中，所指之編碼一或更多所關注之蛋白質的核酸序列包括編碼抗原、免疫調節因子或佐劑之任何組合的至少一傳遞者核苷酸序列。
  34. 如申請專利範圍第23項之組成物，其中，所指之組成物為疫苗。
TW094141904A 2004-12-01 2005-11-29 具有增強之逃脫核內體能力的重組卡介苗（ｂｃｇ）菌株 TWI456057B (zh)

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