Консервативный конформационный эпитоп в стержневом регионе гемагглютинина вирусов гриппа А филогенетически близких подтипов H1, H2, H5 и H6
Поверхностный гликопротеид гемагглютинин (НА) является основным антигенным компонентом вирусов гриппа. Вируснейтрализующие антитела, индуцированные гемагглютинином, составляют основу гуморального иммунитета, защищающего организм человека от гриппозной инфекции. Антигенная структура гемагглютинина подвержена постоянным изменениям в результате селективного давления иммунной системы… Читать ещё >
Содержание
- Список употребляемых сокращений
- I. Введение 5 П. Обзор литературы
- ПЛ. Общее строение вириона вируса гриппа и функции вирусных 10 белков
- П. 2. Гемагглютинин вирусов гриппа А, его структура и функции
- II. 2.1. Рецептор связывающий сайт 20 П. 2.2. Пептид слияния
- 11. 2. 3. Участок нарезания гемагглютинина (протеолитисеский сайт)
- 11. 2. 4. Антигенная структура гемагглютинина 27 П. З. Изменения гемагглютинина в процессе адаптации вирусов гриппа к новому хозяину (на примере адаптации к легким мышей)
- III. Материалы и методы
- III. 1. Вирусы
- Ш. 2. Клетки
- Ш. З. Моноклональное антитело С
- Ш. 4. Реакция торможения гемагглютинации (РТГА)
- Ш. 5. Нейтрализация инфекционности вирусов в культуре клеток
- Ш. 6. Пассирование вируса в легких мышей
- 111. 7. Титрование инфекционности вирусов на куриных эмбрионах
- 111. 8. Титрование инфекционности вирусов на мышах
- 111. 9. Получение радиоактивно меченых вирусспецифических белков 48 для реакции радиоиммунопреципитации
- ШЛО. Получение вирусспецифических белков, радиоактивно меченых в пульс — чейз условиях. 111.11. Радиоактивное мечение вирусспецифических белков в присутствии туникамицина
- ШЛ2. Реакция радиоиммунопреципитации (РИП)
- 111. 13. Электрофорез белков в полиакриламидном геле
- II. L14. Определение нуклеотидной последовательности гена НА
- 111. 15. Компьютерный анализ нуклеотидных последовательностей 56 генов НА
- 111. 16. Сравнение аминокислотных последовательностей НА 56 различных штаммов и анализ локализации замен на 3-мерной структуре НА подтипа НЗ
- 111. 17. Пассивная иммунизация мышей МКА С179 и 56 экспериментальная инфекция
- IV. Результаты
IV.1. Взаимодействие МКА С179 с вирусами гриппа подтипа Н5 и Н6 58 IV.1.1. Нейтрализация вирусов подтипов Н5 и Н6 моноклональным антителом С179 в культуре клеток IV. 1.2. Распознавание НА подтипов Н5 и Нб МКА С179 в реакции радиоиммунопреципитации (РИП) IV.1.3. Влияние созревания НА подтипа Н6 на его распознавание МКА 60 С179 в РИП
IV. 1.4. Взаимодействие МКА С179 с НА подтипа Н6, меченным в присутствии туникамицина, в РИП rV.1.5. Сравнение аминокислотных последовательностей участков, распознаваемых МКА С179, в НА подтипов HI, Н2, Н5, и Нб IV.1.6. Потенциальные сайты гликозилирования в НА подтипов HI, 69 Н2, Н5 и Нб
IV.2. Адаптация птичьего штамма вируса гриппа A (H5N2) к легким 69 мышей.
IV.2.1. Репродукция вируса в легких мышей в ходе адаптации.
Вирулентность адаптированного варианта Mal. dk/PA/84-MA IV.2.2. Антигенные свойства НА вирусов Mal. dk/PA/84 и 73 Mal. dk/PA/84-MA. Резистентность к а- и р-ингибиторам гемагглютинации
IV.2.3. Анализ первичной структуры НА вирусов Mal. dk/PA/84 и 74 Mal. dk/PA/84-MA
IV.2.4. Реакции in vitro МКА С179 с адаптированными к мышам вариантами птичьих штаммов с подтипами НА Н5 и Н2 IV.3. Эксперименты in vivo по профилактике и лечению летальной бронхопневмонии мышей, вызванной вирусами гриппа Hl, Н2 и Н5 подтипов, моноклональным антителом С
1У.3.1. Профилактический эффект МКА С179 при инфекции, 79 вызываемой вирусом гриппа, А человека с НА подтипа Н
IV.3.2. Профилактический и лечебный эффекты МКА С179 при 80 инфекции, вызываемой вирусом гриппа птиц с НА подтипа Н
IV.3.3. Профилактический и лечебный эффекты МКА С179 при 81 инфекции, вызываемой вирусом гриппа птиц с НА подтипа Н
V. Обсуждение
VI. Выводы
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