Изучение экспрессии генов при дифференцировке и созревании дендритных клеток человека
Диссертация
В нашей работе впервые показана повышенная экспрессия как ФНО-ос в нДК (в 40 раз в нДК, но сравнению с моноцитами), так и его рецептора II (в 10.2 раза). Дифференциальная интенсивность сигналов, полученных гибридизацией на макроматрице, для ФНО-а была подтверждена методом полуколичественной ПЦР на матрице кДНК четырех доноров. Эти результаты интересны тем, что показывают '" включение" генов… Читать ещё >
Содержание
- ГЛАВА 1. ОБЗОР ЛИТЕРАТУРЫ
- 1. 1. Определение, классификация и функции дендритных клеток
- 1. 1. 1. Типы дендритных клеток
- 1. 1. 2. Фенотип
- 1. 1. 2. 1. Морфология
- 1. 1. 2. 2. Поверхностные антигены
- 1. 1. 3. Функции дендритных клеток
- 1. 1. 3. 1. Иммунный ответ
- 1. 1. 3. 2. Толерантность к собственным антигенам
- 1. 1. Определение, классификация и функции дендритных клеток
- 1. 2. Дендритные клетки в клинической иммунологии
- 1. 3. Дендритные клетки и В-лимфоциты
- 1. 4. Созревание дендритных клеток
- 1. 5. Методы изучения дендритных клеток
- 1. 5. 1. История
- 1. 5. 2. Получение дендритных клеток in vitro
- 1. 5. 3. Методы изучения экспрессии генов в дендритных клетках
- 1. 5. 3. 1. Секвенирование библиотек кДНК
- 1. 5. 3. 2. SAGE метод
- 1. 5. 3. 3. Дифференциальный дисплей мРНК
- 1. 5. 3. 4. Гибридизация на кДНК матрицу
- 2. 1. Получение дендритных клеток in vitro
- 2. 2. Фенотипирование дендритных клеток моноклональными антителами
- 2. 3. Выделение суммарной РНК и обработка ДНКазой
- 2. 4. Обратная транскрипция и амплификация мишеней для макроматриц
- 2. 5. Мечение кДНК мишеней для анализа па макроматрице
- 2. 6. Гибридизация, детекция и количественный анализ макроматриц
- 2. 7. Мечение кДНК мишеней для микроматриц
- 2. 8. Гибридизация на микроматрицу и количественный анализ
- 2. 9. Полуколичественная полимеразная цепная реакция на матрице кДНК
- 2. 10. Прямое секвенирование нуклеотидных последовательностей фрагментов кДНК и поиск гомологии
- 3. 1. Получение in vitro незрелых и зрелых дендритных клеток человека. Фенотипирование поверхностных маркеров
- 3. 2. Мониторинг экспрессии генов в моноцитах и незрелых дендритных клетках методом гибридизации на макроматрице
- 3. 3. Подтверждение профилей гибридизации на кДНК макроматрицу методом полуколичественной ПЦР на матрице кДНК
- 3. 4. Мониторинг экспрессии генов в незрелых и зрелых дендритных клетках методом гибридизации на микроматрицу
- 3. 5. Подтверждение профилей гибридизации на кДНК микроматрице методом полуколичественной Г11 (Р на матрице кДНК
- 3. 6. Экспрессия генов ренин-ангиотензиновой системы
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