Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes: = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Würzburg
2017
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Volltext Inhaltsverzeichnis |
| Beschreibung: | 204 Seiten Illustrationen, Diagramme 21 cm |
Internformat
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| 100 | 1 | |a Garcia Guerrero, Estefania |d 1987- |e Verfasser |0 (DE-588)1142033139 |4 aut | |
| 245 | 1 | 0 | |a Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes |b = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |c submitted by Estefania Garcia Guerrero from Seville-Spain |
| 246 | 1 | 1 | |a Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| 264 | 1 | |a Würzburg |c 2017 | |
| 300 | |a 204 Seiten |b Illustrationen, Diagramme |c 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Julius-Maximilians-University Würzburg and University of Seville |d 2017 | ||
| 546 | |a Zusammenfassung in deutscher und englischer Sprache | ||
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| 650 | 0 | 7 | |a Krebs |g Medizin |0 (DE-588)4073781-0 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a T-Lymphozyt |0 (DE-588)4127387-4 |2 gnd |9 rswk-swf |
| 653 | |a Strategies to Obtain Tumor-Reactive Cells | ||
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| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:20-opus-150547 |a Garcia Guerrero, Estefania, 1987- |t Strategies to Obtain Tumor-Reactive Cells for Cancer Immunotherapy by Cell Sorting and Genetic Modifications of T Lymphocytes |d Würzburg : Universität Würzburg, 2017 |h Online-Ressource |
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Datensatz im Suchindex
| _version_ | 1804178405248204800 |
|---|---|
| adam_text | ABBREVIATIONS
17
A BSTRA
CT.................................................................................................
21
ZUSAM M ENFASSUNG (G ERM AN)
.......................................................
23
IN TRO DU CTIO N
........................................................................................25
1. ADOPTIVE T CELL IMMUNOTHERAPY FOR CANCER
...............................
27
2. CYTOTOXIC T CELLS: RECOGNITION OF TUMOR ANTIGENS
...................
32
3. CONVENTIONAL METHODS FOR OBTAINING TUMOR-SPECIFIC CYTOTOXIC
T
CELLS.......................................................................................
35
4. THE T CELL-TUMOR CELL INTERACTION
.................................................
38
5. TUMOR-INFILTRATING LYMPHOCYTE THERAPY
.......................................
41
6. GENE-MODIFIED T CELL THERAPY
......................................................
45
7. CAR DESIGN AND MODE OF ACTION
..................................................
48
8. CAR CLINICAL
TRIALS............................................................................
54
H YPO TH ESIS
............................................................................................
55
O B
JECTIVES.............................................................................................59
MATERIAL AND M E TH O D S
.................................................................
63
1. BIOLOGICAL MATERIAL
............................................................................
65
1.1. HEALTHY HUMAN
SAMPLES...................................................65
1.2. TUMOR HUMAN
SAMPLES.....................................................65
1.2.1. ACUTE MYELOID LEUKEMIA (AML)
.............................
65
1.2.2. MULTIPLE MYELOMA (M M )
..........................................
66
1.3. TUMOR CELL LINES
.................................................................
66
1.3.1. MULTIPLE MYELOMA CELL LINES
....................................
66
1.3.2. OTHER CELL LINES
..........................................................
67
2. NON-BIOLOGICAL MATERIAL
.....................................................................
69
LL
2 .1. EQUIPMENT AND CONSUMABLES
..........................................
69
2.2.
SOFTWARE.................................................................
71
2.3. CHEMICALS AND REAGENTS
..................................................
71
2.3.1. MOLECULAR BIOLOGY
.....................................................
71
2.3.2. CELL CULTURE AND IMMUNOLOGY.................................72
2.4. MEDIA AND
BUFFERS................................................................74
2.5. COMMERCIAL KITS
..................................................................
76
2.6. ANTIBODIES
.............................................................................
76
2.6.1. FLOW
CYTOMETRY.......................................................... 76
2.6.2. WESTERN BLOT
............................................................
78
3. MOLECULAR
DYNAMICS...........................................................................79
4. DOUBLET CELL CULTURE AND FUNCTIONAL TESTS
...................................
81
4.1. PBMC
PURIFICATION................................................................81
4.2. CD3 DEPLETION AND IRRADIATION
..........................................
81
4.3. PRIMARY CO-CULTURE
.............................................................
82
4.4. FACS-BASED CELL SORTING
..................................................
83
4.5. IMMUNOPHENOTYPE
...............................................................
84
4.6. SECONDARY
CO-CULTURES......................................................85
4.6.1. CYTOTOXICITY ASSAY
....................................................
85
4.6.2. SUPPRESSION
ASSAY...................................................85
4.6.3. ACTIVATION ASSAY
........................................................
86
5. CAR MANUFACTURING AND FUNCTIONAL TESTS
....................................
88
5.1. PREPARATION OF BCMA CAR-ENCODING PLASMIDS
...........
88
5.2. PREPARATION OF VIRAL
VECTORS..............................................89
5.3. TITRATION OFLENTIVIRUS
..........................................................
90
5.4. ISOLATION OF HUMAN T CELL
SUBSETS..................................91
5.5. LENTIVIRAL TRANSDUCTION OF T CELLS
....................................
92
5.6. ENRICHMENT OF CAR+ T CELLS
.............................
92
5.7. ANTIGEN DEPENDENT EXPANSION
........................................
93
5.8. IMMUNOLOGICAL AND FUNCTIONAL TESTS
..............................
93
5.8.1. IMMUNOPHENOTYPE
...................................................
93
5.8.2. CYTOTOXICITY ASSAY
....................................................
94
5.8.3. CYTOKINE SECRETION ASSAY AND ELISA
..................
94
5.8.4. CFSE PROLIFERATION ASSAY
.......................................
95
5.9. FUNCTIONAL TESTS IN THE PRESENCE OF SOLUBLE
B CM
A.....................................................................................
95
5.9.1. CYTOTOXICITY ASSAY
..........
.......................................95
5.9.2. ACTIVATION
ASSAY........................................................ 96
6. WESTERN
BLOTTING..................................................................................97
7. PRECLINICAL IN VIVO
EXPERIMENTS............................................ 99
7 .1. MULTIPLE MYELOMA XENOGRAFT MODEL
.................................
99
7.2. ADOPTIVE TRANSFER OF T CELLS AND ANALYSIS OF ANTITUMOR
EFFICACY...................................................................................99
8. STATISTICAL ANALYSIS
...........................................................................
100
RESULTS..................................................................................................101
PART I: *DOUBLET TEC H NO LO G Y* TO OBTAIN TUM OR
REACTIVE T CELLS
1. REACTIVITY OF PMHC COMPLEXES CORRELATES WITH THE STRENGTH OF
PMHC-TCR INTERACTION
..................................................................
105
1.1. MODEL
SYSTEM.....................................................................105
1.2. STABILITY OF COMPUTATIONAL SIMULATIONS
...........................
106
1.3. EFFECT OF THE PEPTIDE IN HLA STRUCTURE
...........................
109
1.4. EFFECTS OF TCR BINDING ON PHLA DYNAMICS..................112
1.5. SALT-BRIDGE PATTERNS AND ELECTROSTATICS
........................
114
2. THE BASICS OF *DOUBLET TECHNOLOGY*: CO-CULTURE, INCUBATION
TIME AND FACS-BASED CELL SORTING
...........................................
122
2.1. CO-CULTURE CONDITIONS
........................................................
122
2.2. INCUBATION
TIME...................................................................
125
2.3. FACS-BASED CELL
SORTING................................................126
DOUBLET T CELLS SHOW HIGHER PERCENTAGE OF EFFECTOR CELLS AND
SPECIFIC CYTOTOXIC ACTIVITY AS COMPARED TO NON-DOUBLET T
CELLS...............................................................................
.................
128
3 .1.
IMMUNOPHENOTYPE............................................................
128
3.2. CYTOTOXIC
ACTIVITY...............................................................
132
A SUBSET OF NON-DOUBLET T CELLS HAVE IMMUNO-SUPPRESSIVE
FUNCTION.............................................................................................135
4.1.
IMMUNOPHENOTYPE............................................................135
4.2. IMMUNOSUPPRESSIVE
FUNCTION.........................................135
CLINICAL APPLICATION: DOUBLET T CELLS FROM AML PATIENTS HAVE
SPECIFIC CYTOTOXIC ACTIVITY AGAINST PRIMARY BLAST CELLS
...........
138
5 .1. PATIENT
SAMPLES..................................................................138
5.2. CYTOTOXIC ACTIVITY OF DOUBLET T CELLS AGAINST BLAST
CELLS.................................................. 139
INTERIM CONCLUSION FOR *DOUBLET TECHNOLOGY*
...........................
143
PART II: CAR TEC H NO LO G Y TO GENERATE TUM OR
REACTIVE T CELLS
1. SUBSTANTIAL EXPRESSION OF BCMA ON MYELOMA CELLS AND
GENERATION OF BCMA CAR T CELLS
.............................................
147
1.1. BCMA EXPRESSION ON MYELOMA CELLS
............................
147
1.2. BCMA EXPRESSION ON NON-MYELOMA CELLS
....................
148
1.3. CAR DESIGN: 2 GENERATION CAR WITH 4-1BB
DOMAIN..................................................................................149
1.4. TRANSDUCTION OF T CELLS BY LENTIVIRAL GENE TRANSFER..... 152
2. BCMA CAR DESIGN AFFECTS ANTI-MYELOMA FUNCTION
..............
156
3. BCMA CAR T CELLS ELIMINATE MYELOMA CELLS IN VITRO
............
157
3.1. CYTOTOXIC ACTIVITY OF BCMA CAR T CELLS
.......................
157
3.2. CYTOKINE PRODUCTION OF BCMA CAR T CELLS
.................
158
3.3. PROLIFERATION OF BCMA CAR T CELLS.............................159
4. THE SERUM OF MULTIPLE MYELOMA PATIENTS CONTAINS A SOLUBLE
FORM OF BCM A WHICH IS CORRELATED WITH DISEASE STATUS
.....
161
5. SOLUBLE BCMA DOES NOT ABROGATE THE EFFICACY OF BCMA CAR
T
CELLS................................................................................................163
5.1. CYTOTOXIC ACTIVITY IN THE PRESENCE OF SBCMA...............163
5.2. ACTIVATION IN THE PRESENCE OF SBCMA
...........................
164
5.3. SBCMA PROTEIN DETECTION
................................................
165
6. BCMA CAR T CELLS ERADICATE TUMOR IN VIVO
...........................
167
7. INTERIM CONCLUSION FOR CAR TECHNOLOGY
...................................
167
D ISCU SSIO N
...........................................................................................171
1. *DOUBLET TECHNOLOGY* TO OBTAIN TUMOR-REACTIVE T CELLS FROM
ACUTE MYELOID LEUKEMIA PATIENTS
..................................
173
2. CAR TECHNOLOGY TO GENERATE TUMOR-REACTIVE T CELLS FROM
MULTIPLE MYELOMA PATIENTS
............................................................
178
3. CLINICAL
TRANSLATION..........................................................................
183
4. FINAL REMARKS AND FURTHER W
ORK..................................................166
C O N CLUSIO N
S.......................................................................................187
R EFEREN
CES.........................................................................................191
CURRICULUM V IT A E
...........................................................................205
|
| any_adam_object | 1 |
| author | Garcia Guerrero, Estefania 1987- |
| author_GND | (DE-588)1142033139 |
| author_facet | Garcia Guerrero, Estefania 1987- |
| author_role | aut |
| author_sort | Garcia Guerrero, Estefania 1987- |
| author_variant | g e g ge geg |
| building | Verbundindex |
| bvnumber | BV044872905 |
| collection | ebook |
| ctrlnum | (OCoLC)1017028692 (DE-599)DNB1142033309 |
| discipline | Medizin |
| format | Thesis Book |
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| id | DE-604.BV044872905 |
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| institution | BVB |
| language | English |
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| physical | 204 Seiten Illustrationen, Diagramme 21 cm |
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| publishDateSort | 2017 |
| record_format | marc |
| spelling | Garcia Guerrero, Estefania 1987- Verfasser (DE-588)1142033139 aut Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie submitted by Estefania Garcia Guerrero from Seville-Spain Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie Würzburg 2017 204 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Julius-Maximilians-University Würzburg and University of Seville 2017 Zusammenfassung in deutscher und englischer Sprache Herstellung (DE-588)4159653-5 gnd rswk-swf Immuntherapie (DE-588)4026640-0 gnd rswk-swf Krebs Medizin (DE-588)4073781-0 gnd rswk-swf T-Lymphozyt (DE-588)4127387-4 gnd rswk-swf Strategies to Obtain Tumor-Reactive Cells (DE-588)4113937-9 Hochschulschrift gnd-content Krebs Medizin (DE-588)4073781-0 s Immuntherapie (DE-588)4026640-0 s T-Lymphozyt (DE-588)4127387-4 s Herstellung (DE-588)4159653-5 s DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:20-opus-150547 Garcia Guerrero, Estefania, 1987- Strategies to Obtain Tumor-Reactive Cells for Cancer Immunotherapy by Cell Sorting and Genetic Modifications of T Lymphocytes Würzburg : Universität Würzburg, 2017 Online-Ressource B:DE-101 application/pdf http://d-nb.info/1142033309/04 Inhaltsverzeichnis https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-150547 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030267305&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Garcia Guerrero, Estefania 1987- Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie Herstellung (DE-588)4159653-5 gnd Immuntherapie (DE-588)4026640-0 gnd Krebs Medizin (DE-588)4073781-0 gnd T-Lymphozyt (DE-588)4127387-4 gnd |
| subject_GND | (DE-588)4159653-5 (DE-588)4026640-0 (DE-588)4073781-0 (DE-588)4127387-4 (DE-588)4113937-9 |
| title | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| title_alt | Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| title_auth | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| title_exact_search | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| title_full | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie submitted by Estefania Garcia Guerrero from Seville-Spain |
| title_fullStr | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie submitted by Estefania Garcia Guerrero from Seville-Spain |
| title_full_unstemmed | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie submitted by Estefania Garcia Guerrero from Seville-Spain |
| title_short | Strategies to obtain tumor-reactive cells for cancer immunotherapy by cell sorting and genetic modifications of T lymphocytes |
| title_sort | strategies to obtain tumor reactive cells for cancer immunotherapy by cell sorting and genetic modifications of t lymphocytes zellsortierung und genetische modifikation von t lymphozyten zur gewinnung tumorreaktiver zellen fur die krebsimmuntherapie |
| title_sub | = Zellsortierung und genetische Modifikation von T-Lymphozyten zur Gewinnung tumorreaktiver Zellen für die Krebsimmuntherapie |
| topic | Herstellung (DE-588)4159653-5 gnd Immuntherapie (DE-588)4026640-0 gnd Krebs Medizin (DE-588)4073781-0 gnd T-Lymphozyt (DE-588)4127387-4 gnd |
| topic_facet | Herstellung Immuntherapie Krebs Medizin T-Lymphozyt Hochschulschrift |
| url | http://d-nb.info/1142033309/04 https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-150547 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030267305&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT garciaguerreroestefania strategiestoobtaintumorreactivecellsforcancerimmunotherapybycellsortingandgeneticmodificationsoftlymphocyteszellsortierungundgenetischemodifikationvontlymphozytenzurgewinnungtumorreaktiverzellenfurdiekrebsimmuntherapie AT garciaguerreroestefania zellsortierungundgenetischemodifikationvontlymphozytenzurgewinnungtumorreaktiverzellenfurdiekrebsimmuntherapie |
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