RU2009115177A - Живые бактериальные вакцины для профилактики и лечения вирусной инфекции - Google Patents

Живые бактериальные вакцины для профилактики и лечения вирусной инфекции Download PDF

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RU2009115177A
RU2009115177A RU2009115177/15A RU2009115177A RU2009115177A RU 2009115177 A RU2009115177 A RU 2009115177A RU 2009115177/15 A RU2009115177/15 A RU 2009115177/15A RU 2009115177 A RU2009115177 A RU 2009115177A RU 2009115177 A RU2009115177 A RU 2009115177A
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Дэвид Гордон БЕРМЬЮДЕЗ (CA)
Дэвид Гордон БЕРМЬЮДЕЗ
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Abstract

1. Состав живой вакцины для защиты животного от заражения птичьим гриппом, содержащий живую аттенюированную бактерию Сальмонелла, содержащий: ! аттенюирующую мутацию в генетическом локусе хромосомы указанной бактерии, которая ослабляет вирулентность указанной бактерии; ! антиген-выражающую конструкцию ДНК, содержащую: ! нуклеотидную последовательность, кодирующую иммуногенный полипептид, содержащий антиген птичьего гриппа H или N, ! иммуногенную часть антигена H или N или; ! их комбинацию, ! где вышеуказанная нуклеотидная последовательность функционально связана с промотором, обеспечивающим выражение указанного иммуногенного полипептида с указанной конструкции ДНК; ген, кодирующий иммуногенный полипептид, имеющий по крайней мере один кодон, оптимизированный для выражения в бактериях, и указанный живой состав вакцины вызывает иммунную реакцию к по меньшей мере одному антигену птичьего гриппа при пероральном введении животному. ! 2. Живой состав вакцины по п.1, где указанные живые аттенюированные бактерии Сальмонелла выбраны из группы, состоящей из Salmonella enterica serovar Typhimuriurn (S. Typhimuriurn), Salmonella enterica serovar Typhi (S. typhi), 34 Salmonella enterica serovar Paratyphi B (S. paratyphi B), Salmonella enterica serovar Paratyphi C (S. paratyphi Q), Salmonella enteric serovar Hadar (S. Hadar), Salmonella enterica serovar Enteriditis (S. Enteriditis), Salmonella enterica serovar Kentucky (S. kentucky), Salmonella enteric serovar Infantis (S. infantis), Salmonella enterica serovar Pullorum (S. pullorum), Salmonella enterica serovar Gallinarum (S. gallinarum), Salmonella enterica serovar Muenchen (S. muenchen), 25 Salmonella enterica serovar Anatum (S. anatum), Salmonella enterica serovar Dublin (S. dublin), Salmonella enterica serovar Derby (S. derby), Salmonella enterica serovar Choleraesuis var. kunzendorf, и Salmonella enterica serovar Minnesota. ! 3. Живой состав вакцины по п.2, где указанной живой аттенюированной бактерией Сальмонеллой является S.enterica sero

Claims (25)

1. Состав живой вакцины для защиты животного от заражения птичьим гриппом, содержащий живую аттенюированную бактерию Сальмонелла, содержащий:
аттенюирующую мутацию в генетическом локусе хромосомы указанной бактерии, которая ослабляет вирулентность указанной бактерии;
антиген-выражающую конструкцию ДНК, содержащую:
нуклеотидную последовательность, кодирующую иммуногенный полипептид, содержащий антиген птичьего гриппа H или N,
иммуногенную часть антигена H или N или;
их комбинацию,
где вышеуказанная нуклеотидная последовательность функционально связана с промотором, обеспечивающим выражение указанного иммуногенного полипептида с указанной конструкции ДНК; ген, кодирующий иммуногенный полипептид, имеющий по крайней мере один кодон, оптимизированный для выражения в бактериях, и указанный живой состав вакцины вызывает иммунную реакцию к по меньшей мере одному антигену птичьего гриппа при пероральном введении животному.
2. Живой состав вакцины по п.1, где указанные живые аттенюированные бактерии Сальмонелла выбраны из группы, состоящей из Salmonella enterica serovar Typhimuriurn (S. Typhimuriurn), Salmonella enterica serovar Typhi (S. typhi), 34 Salmonella enterica serovar Paratyphi B (S. paratyphi B), Salmonella enterica serovar Paratyphi C (S. paratyphi Q), Salmonella enteric serovar Hadar (S. Hadar), Salmonella enterica serovar Enteriditis (S. Enteriditis), Salmonella enterica serovar Kentucky (S. kentucky), Salmonella enteric serovar Infantis (S. infantis), Salmonella enterica serovar Pullorum (S. pullorum), Salmonella enterica serovar Gallinarum (S. gallinarum), Salmonella enterica serovar Muenchen (S. muenchen), 25 Salmonella enterica serovar Anatum (S. anatum), Salmonella enterica serovar Dublin (S. dublin), Salmonella enterica serovar Derby (S. derby), Salmonella enterica serovar Choleraesuis var. kunzendorf, и Salmonella enterica serovar Minnesota.
3. Живой состав вакцины по п.2, где указанной живой аттенюированной бактерией Сальмонеллой является S.enterica serovar Typhimurium (S.typhimurium).
4. Живой состав вакцины по п.1, где указанная аттенюированная мутация находится в генетическом локусе, выбранном из группы, состоящей из phoP, phoQ, Mt, cya, crp, poxA, rpoS, htrA, nuoG, pmi, galE, pabA, pts, damA, purA, purB, purI, zwf, gua, cadA, rfic, rjb, rfa, ompR, Suwwan, а также их комбинации.
5. Живой состав вакцины по п.4, где аттенюирующей мутацией является мутация делеции.
6. Живой состав вакцины по п.5, где аттенюирующая мутация содержит по крайней мере мутацию частичной делеции phoP.
7. Живой состав вакцины по п.1, где указанная бактерия Сальмонелла содержит летальную мутацию, содержащую делецию в гене asd, и указанный иммуногенный полипептид содержит слитый белок, содержащий антиген V или его иммуногенную часть, связанную с антигеном F1 или его иммуногенной частью, кодируемым антиген-экспрессирующим мультикопированном плазмидом.
8. Живой состав вакцины по п.7, где бэкграундом воспроизведения указанного мультикопированном плазмиде является ColE1, pUC, M15, or pBR322.
9. Живой состав вакцины по п.1, где указанный живой состав вакцины содержит множество живых сероваров Сальмонеллы.
10. Живой состав вакцины по п.1, где указанная живая аттенюированная бактерия Сальмонелла генетически стабилизирована по сравнению с диким типом Сальмонелла того же серовара.
11. Живой состав вакцины по п.1, где указанная живая аттенюированная бактерия Сальмонелла генетически стабилизирована против генетического обмена с другими организмами по сравнению с диким типом Сальмонеллы того же серовара.
12. Живой состав вакцины содержащий бактерию Сальмонеллу, которая выражает по крайней мере один антиген птичьего гриппа H и N, а так же иммуногенную часть указанного Н или N антигена.
13. Живой состав вакцины по п.12, где бактерия Сальмонелла содержит S.typhirmurium.
14. Живой состав вакцины по п.1, полученный из набора, содержащего (а) первый контейнер, содержащий оптимизированный кодоном антиген бактериальной экспрессии патогенного штамма птичьего гриппа, в который входят уникальные генетически произведённые сайты рестрикции, входящие по крайней мере в один из плазмидов бактериальной белковой экспрессии или бактериального хромосомного вектора белковой экспрессии, что позволяет быстрый обмен малых сегментов; и (б) второй контейнер, составляющий бактериальные флагелларные векторы, имеющие, по крайней мере, один флагелларный антиген, где бактерия Сальмонеллы составляет множество уникальных хромосомных векторов локализации, направленных, по меньшей мере, на IS200, элементы фагов и метаболические гены для введения кодона выражения.
15. Способ иммунизации животного против птичьего гриппа, включающий введение живого состава вакцины, который содержит бактерию Сальмонеллу, выражающую по крайней мере один из антигенов H и N птичьего гриппа, и иммуногенную часть этого антигена.
16. Способ по п.15, где состав живой вакцины адаптирован, чтобы защитить животное от заражения птичьим гриппом, где бактерия Сальмонелла содержит аттенюирующую мутацию в генетическом локусе хромосомы указанной бактерии, которая ослабляет вирулентность указанной бактерии; антигено-выражающая конструкция ДНК, содержащая кодирование последовательности нуклеотидов для иммуногенного полипептида, содержащего антиген птичьего гриппа H и N, иммуногенную часть этого антигена H и N или их комбинацию, где указанная последовательность нуклеотидов связана операбельным путем с промотером, который позволяет выражение вышеупомянутого иммуногенного полипептида от вышеупомянутой конструкции ДНК; ген, кодирующий иммуногенный полипептид имеет, по крайней мере, один кодон, оптимизированный для бактериальной экспрессии, и вышеупомянутый состав вакцины выявляет иммунную реакцию хотя бы к одному антигену птичьего гриппа при введении животному оральным путем.
17. Способ по п.16, где бактерия Сальмонелла выбрана из группы, состоящей из Salmonella enterica serovar Typhimuriurn (S. typhimurium), Salmonella enterica serovar Typhi (S. typhi), 34 Salmonella enterica serovar Paratyphi B (S. paratyphi B), Salmonella enterica serovar Paratyphi C (S. paratyphi Q, Salmonella enterica serovar Hadar (S. hadar), Salmonella enterica serovar Enteriditis (S. enteriditis), Salmonella enterica serovar Kentucky (S. kentucky), Salmonella enterica serovar Infantis (S. infantis), Salmonella enterica serovar Pullorum (S. pullorum), Salmonella enterica serovar Gallinarum (S. gallinarum), Salmonella enterica serovar Muenchen (S. muenchen), Salmonella enterica serovar Anatum (S. anatum), Salmonella enterica serovar Dublin (S. dublin), Salmonella enterica serovar Derby (S. derby), and Salmonella enterica serovar Choleraesuis var. kunzendorf, и Salmonella enterica serovar Minnesota.
18. Способ по п.17, где бактерия Сальмонеллы содержит S.enterica serovar Typhimurium (S. typhimurium).
19. Способ по п.16, где аттенюирующая мутация находится в генетическом локусе, выбранном из группы, состоящей из phoP, phoQ, Mt, cya, crp, poxA, rpoS, htrA, nuoG, pmi, galE, pabA, pts, damA, purA, purB, purI, zwf, gua, cadA, rfic, rjb, rfa, ompR, Suwwan и их комбинации.
20. Способ по п.19, где аттенюирующая мутация содержит по крайней мере частичную делецию phoP.
21. Способ по п.16, где бактерия Сальмонелла содержит летальную мутацию, содержащую делецию гена asd, и указанный иммуногенный полипептид содержит слитый белок, содержащий антиген V или его иммуногенную часть, связанную с антигеном F1 или с его иммуногенной частью, кодируемой на антигено-выражающем, мультикопированном плазмиде.
22. Способ по п.16, где бактерия Сальмонелла генетически стабилизирована по сравнению с диким типом Сальмонеллы того же серовара.
23. Способ по п.16, где живая аттенюированная бактерия Сальмонелла генетически стабилизирована путем делеции элементов IS200 и бактериальных элементов фагов и профагов, и генетически изолирована от внешнего фагового заражения путем составного выражения репрессора фага P22.
24. Набор, адаптированный для использования в получении живого состава вакцины, содержащего бактерию Сальмонелла, которая выражает по крайней мере один из антигенов H и N птичьего гриппа, и иммуногенную часть указанного антигена H и N, содержащий: (а) первый контейнер, содержащий антиген оптимизированный кодоном бактериальной экспрессии против патогенного штамма птичьего гриппа, в который входят уникальные генетически произведенные сайты рестрикции, входящие по крайней мере в один из бактериального плазмида белковой экспрессии или вектор экспрессии бактериального хромосомного белка; и (б) второй контейнер, составляющий бактериальные флагелларные векторы, имеющие по крайней мере один флагелларный антиген.
25. Набор по п.24, дополнительно содержащий бактериальный штамм, где кодон бактериального выражения позволяет быстрый обмен малых сегментов, где бактериальный штамм составляет множество уникальных хромосомных векторов локализации направленных по меньшей мере на IS200, элементы фагов и на метаболические гены для введения кодона экспрессии.
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