Buschow Ch. In-vivo imaging of an inducible oncogenic tumor antigen visualizes tumor progression and predicts cytotoxic T lymphocyte tolerance (Berlin, 2009). - ОГЛАВЛЕНИЕ / CONTENTS
Навигация

Архив выставки новых поступлений | Отечественные поступления | Иностранные поступления | Сиглы
ОбложкаBuschow Ch. In-vivo imaging of an inducible oncogenic tumor antigen visualizes tumor progression and predicts cytotoxic T lymphocyte tolerance: Diss. … Dr. rer. nat. - Berlin: Freie Univ., 2009. - 69 p.: ill., graph. - Ref.: p.54-62.
 

Оглавление / Contents
 
1  Introduction ................................................. 4
   1.1  Mouse models for tumor immunology research .............. 4
        1.1.1  Oncogene transgenic mouse models ................. 4
        1.1.2  SV40 Large T antigen as transgenic oncogene in
               mouse models ..................................... 5
        1.1.3  Immunological characteristics of Tag ............. 5
        1.1.4  Conditional mouse tumor models ................... 6
        1.1.5  Controlled tumor development following Cre4oxP
               mediated recombination ........................... 7
        1.1.6  Leakiness of LoxP-flanked attenuators ............ 9
        1.1.7  The LoxP-Tag transgenic mouse model .............. 9
   1.2  Small Animal Imaging ................................... 11
        1.2.1  Firefly luciferase for in vivo imaging .......... 13
        1.2.2  Mouse models for bioluminescent imaging ......... 14
   1.3  Adoptive T cell therapy ................................ 16
   1.4  Aim of this work ....................................... 17
2  Material and Methods ........................................ 18
   2.1  Restriction digestion .................................. 18
   2.2  Gel electrophoresis and gel extraction ................. 18
   2.3  Ligation reaction ...................................... 18
   2.4  Propagation of plasmids in bacteria .................... 18
   2.5  Purification of plasmids ............................... 19
   2.6  Cell culture ........................................... 19
   2.7  Construction of the TagLuc fusion gene ................. 19
   2.8  Plasmids for analysis of transformation and
        luciferase activity of the TagLuc fusion protein ....... 20
   2.9  Analysis of transformation and luciferase activity of
        the TagLuc fusion protein .............................. 20
   2.10 Western blot analysis .................................. 20
   2.11 Plasmids for generation of transgenic mice ............. 21
   2.12 Analysis of Cre-mediated recombination of
        LoxP-TagLuc-pA and LoxP-TagLuc plasmids ................ 21
   2.13 Mice ................................................... 21
   2.14 PCR .................................................... 22
   2.15 Bioluminescent imaging ................................. 22
   2.16 Histology and immunohistochemistry ..................... 23
   2.17 In vivo cytotoxicity assay ............................. 23
   2.18 ELISA .................................................. 24
   2.19 Adoptive T cell transfer ............................... 24
3  Results ..................................................... 25
   3.1  Design and in vitro analysis of the TagLuc fusion
        gene ................................................... 25
        3.1.1  Western blot analysis of the TagLuc fusion
               protein ......................................... 26
        3.1.2  Oncogenic and luciferase activity of the
               TagLuc protein .................................. 28
   3.2  Development of conditional TagLuc transgenic mice ...... 30
        3.2.1  Generation of LoxP-TagLuc-pA transgenic mice
               and in vivo expression of TagLuc ................ 30
        3.2.2  CTL response towards Tag in LoxP-TagLuc-pA
               transgenic mice ................................. 32
        3.2.3  Generation of LoxP-TagLuc transgenic mice ....... 34
        3.2.4  CTL response towards Tag in LoxP-TagLuc
               transgenic mice ................................. 38
        3.2.5  Cre-independent TagLuc gene expression in
               individual organs from LoxP-TagLuc-pA and
               LoxP-TagLuc transgenic mice ..................... 39
        3.2.6  Analyzing in vitro expression of
               LoxP-TagLuc-pA and LoxP-TagLuc constructs by
               transient transfection .......................... 42
   3.3  Tumor development in LoxP-TagLuc mice .................. 43
        3.3.1  LoxP-TagLuc mice did not develop sporadic
               tumors .......................................... 43
        3.3.2  Detection of hepatocellular carcinoma in
               LoxP-TagLuc transgenic mice crossed to liver-
               specific Alb-Cre-trangenic mice ................. 45
        3.3.3  Ex vivo analysis of the liver tumor from
               LoxP-TagLuc x Alb-Cre double transgenic mice .... 47
   3.4 Evaluating the therapeutic potential of adoptively-
       transferred Tag-specific T cells into LoxP-TagLuc and
       LoxP-TagLuc-pA mice ..................................... 48
4  Discussion .................................................. 50
5  References .................................................. 54
6  Summary ..................................................... 54
7  Zusammenfassung ............................................. 64
8  Acknowledgements ............................................ 65
9  Appendix .................................................... 66
   Plasmid maps ................................................ 66
   LoxP-TagLuc-pA .............................................. 66
   LoxP-TagLuc ................................................. 67
   pCAG-Tag .................................................... 68
   pCAG-TagLuc ................................................. 69


Архив выставки новых поступлений | Отечественные поступления | Иностранные поступления | Сиглы
 

[О библиотеке | Академгородок | Новости | Выставки | Ресурсы | Библиография | Партнеры | ИнфоЛоция | Поиск]
  Пожелания и письма: branch@gpntbsib.ru
© 1997-2024 Отделение ГПНТБ СО РАН (Новосибирск)
Статистика доступов: архив | текущая статистика
 

Документ изменен: Wed Feb 27 14:23:06 2019. Размер: 9,226 bytes.
Посещение N 1455 c 07.02.2012