Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV: = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Erlangen ; Nürnberg
2018
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | viii, 116 Seiten Illustrationen, Diagramme 21 cm |
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| 100 | 1 | |a Elsayed, Hassan |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV |b = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |c vorgelegt von Hassan Elsayed aus Sharkia, Ägypten |
| 246 | 1 | 3 | |a Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| 246 | 1 | 1 | |a Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| 264 | 1 | |a Erlangen ; Nürnberg |c 2018 | |
| 300 | |a viii, 116 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 Friedrich-Alexander-Universität Erlangen-Nürnberg |d 2018 | ||
| 650 | 0 | 7 | |a Immunreaktion |0 (DE-588)4133841-8 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Virus-like particles |0 (DE-588)1078933014 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a HIV |0 (DE-588)4200792-6 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a T-Lymphozyt |0 (DE-588)4127387-4 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Impfstoff |0 (DE-588)4026655-2 |2 gnd |9 rswk-swf |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 689 | 0 | 0 | |a HIV |0 (DE-588)4200792-6 |D s |
| 689 | 0 | 1 | |a T-Lymphozyt |0 (DE-588)4127387-4 |D s |
| 689 | 0 | 2 | |a Virus-like particles |0 (DE-588)1078933014 |D s |
| 689 | 0 | 3 | |a Immunreaktion |0 (DE-588)4133841-8 |D s |
| 689 | 0 | 4 | |a Impfstoff |0 (DE-588)4026655-2 |D s |
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| 999 | |a oai:aleph.bib-bvb.de:BVB01-030833348 | ||
Datensatz im Suchindex
| _version_ | 1804179342709751808 |
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| adam_text | TABLE OF CONTENTS
PAGE
TABLE OF
CONTENTS....................................................................................................................
I
LIST OF
ABBREVIATIONS............................................................................................................
IV
LIST OF
FIGURES........................................................................................................................
VI
ACKNOWLEDGM
ENT...............................................................................................................
VIII
S UM M
ARY..................................................................................................................................1
ZUSAMMENFASSUNG................................................................................................................3
1
INTRODUCTION........................................................................................................................5
1.1 H IV -1/A ID S
......................................................................................................................
5
1.1.1 DISCOVERY, STRUCTURE AND DIVERSITY
........
1.1.2 INFECTION, PATHOGENESIS AND TREATMENT.
1.1.3 IMMUNE RESPONSE AGAINST INFECTION
....
1.2
VACCINATION................................................................................................................
11
1.2.1 PRINCIPLE OF
VACCINATION..............................................................................................11
1.2.2 LICENSED VACCINE
DEVELOPMENT...............................................................................
11
1.2.3 HIV-1 VACCINE DEVELOPM
ENT...................................................................................13
1.2.3.1 VACCINATION CHALLENGES BY
HIV..............................................................................
13
1.2.3.2 VACCINATION TRIALS AGAINST H IV
................................................................................
14
1.2.4 BROADLY NEUTRALISING ANTIBODIES
(BNABS).................................................................16
1.2.5 VIRUS-LIKE PARTICLES (VLP)
VACCINATION....................................................................17
1.3 INTRASTRUCTURAL H ELP
......................................................................................................19
1.4 AIMS OF THE
STUDY.........................................................................................................20
2 MATERIALS AND
METHODS..................................................................................................
21
2.1 M
ATERIALS.........................................................................................................................21
2.1.1 CHEMICALS, REAGENTS AND
KITS..................................................................................
21
2.1.1.1 CHEMICALS AND
REAGENTS.........................................................................................
21
2.1.1.2
KITS..............................................................................................................................
22
2.1.2
CONSUMABLES..............................................................................................................23
2.1.3
INSTRUMENTS..................................................................................................................23
2.1.4 NUCLEIC
ACIDS..............................................................................................................
24
2.4.1.1
PLASMIDS....................................................................................................................
24
2.4.1.2
OLIGONUCLEOTIDES......................................................................................................
25
2.1.5
PEPTIDES......................................................................................................................
26
2.1.5.1 GAG
PEPTIDES............................................................................................................
26
2.1.5.2 TETANUS
PEPTIDES.....................................................................................................
26
2.1.5.3HBSAG
PEPTIDES........................................................................................................26
2.1.6
STANDARDS....................................................................................................................27
C O - N J O I
2.1.7 PROTEINS, LICENSED VACCINES AND ADJUVANTS
...........................................................
27
2.1.8
ANTIBODIES.....................................................................................................................
27
2.1.9
ENZYMES.......................................................................................................................
28
2.1.10BUFFERS AND M
EDIA......................................................................................................
28
2.1.10.1 BUFFERS AND MEDIA FOR MOLECULAR BIOLOGICAL METHODS
........................................
28
2.1.10.2 BUFFERS AND MEDIA FOR PROTEIN BIOCHEMICAL METHODS
........................................
29
2.1.10.3 BUFFERS AND MEDIA FOR CYTOLOGICAL
METHODS........................................................30
2.1.10.4 BUFFERS AND MEDIA FOR IMMUNOLOGICAL METHODS
.................................................
31
2.1.11
BACTERIA........................................................................................................................
31
2.1.12EUKARYOTIC CELL
LINES....................................................................................................
32
2.1.13ANIMALS..........................................................................................................................32
2.1.14SOFTWARE.......................................................................................................................
32
2.2 M
ETHODS........................................................................................................................
33
2.2.1 MOLECULAR BIOLOGICAL
METHODS...................................................................................
33
2.2.1.1 PLASMID DNA
ISOLATION.............................................................................................
33
2.2.1.2 QUANTITATION OF PLASMID DNA
CONCENTRATIONS.......................................................33
2.2.1.3 DIGESTION OF DNA WITH RESTRICTION
ENZYMES.........................................................34
2.2.1.4 AGAROSE GEL
ELECTROPHORESIS...................................................................................
34
2.2.1.5 GEL EXTRACTION OF DNA
FRAGMENTS...........................................................................
34
2.2.1.6 POLYMERASE CHAIN REACTION
(PCR)..........................................................................
34
2.2.1.7 LIGATION OF DNA
FRAGMENTS......................................................................................
35
2.2.1.8 TRANSFORMATION OF
BACTERIA......................................................................................
35
2.2.2 CYTOLOGICAL
METHODS.................................................................................................36
2.2.2.1 CULTIVATION OF CELL
LINES............................................................................................
36
2.2.2.2 TRANSFECTION OF
CELLS..................................................................................................37
2.2.2.3 PREPARATION OF VIRUS-LIKE
PARTICLES.........................................................................37
2.2.3 PROTEIN BIOCHEMICAL METHODS
..................................................................................
38
2.2.3.1 SDS-POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE).....................................38
2.2 .3 2 COOMASSIE STAINING OF POLYACRYLAMIDE
GELS......................................................... 38
2.2.3.3 SILVER STAINING OF POLYACRYLAMIDE
GELS...................................................................
38
2.2.3.4 WESTERN BLOT
ANALYSES.............................................................................................
39
2.2.3.5 EXPRESSION AND PURIFICATION OF RECOMBINANT ENV CONB GP120
........................
39
2.2.3.6 EXPRESSION AND PURIFICATION OF GST-GAG FUSION PROTEIN
...................................
40
2 2 .3 .1 EXPRESSION AND PURIFICATION OF RECOMBINANT B12 ANTIBODY
SUBTYPES
...............
41
2.2.3.8 DETERMINATION OF PROTEIN CONCENTRATIONS
..............................................................
41
2.2.4 IMMUNOLOGICAL
METHODS.............................................................................................42
2.2.4.1 IMMUNIZATION OF
MICE...............................................................................................
42
2.2.4.2 COLLECTION OF
SERA......................................................................................................
43
2.2.4.3 LYMPHOCYTES
PREPARATION.......................................................................................
43
2.2.4.4
IN VITRO RE-STIMULATION OF LYMPHOCYTES AND INTRACELLULAR CYTOKINE
STAINING
......
43
2.2.4.5 CYTOKINE-SPECIFIC ELISA
..........................................................................................44
2.2.4.6 ANTIGEN-SPECIFIC ANTIBODY
ELISA............................................................................
45
2.2.4.7 IFNY ELISPOT
ASSAY.................................................................................................
46
2.2.4.8 IFNY/IL-4 DUAL ELISPOT
ASSAY.................................................................................47
2.2.4.9
IMMUNOPRECIPITATION................................................................................................
47
2.2.4.10 AVIDITY
ASSAY............................................................................................................48
2.2.4.11 FEY RECEPTOR ACTIVATION
ASSAY..............................................................................
48
2.2.5 ETHICS
STATEMENT.........................................................................................................
49
2.2.6 STATISTICAL
ANALYSIS......................................................................................................
49
3 R
ESULTS...............................................................................................................................50
3.1 PROTEIN PRODUCTION & PURIFICATION
..........................................................................
50
3.1.1 RECOMBINANT ENVGPL 2 0
...........................................................................................50
3.1.2 GST-GAG PROTEIN
(P55)..............................................................................................51
3.1.3 RECOMBINANT B12 IGG
SUBTYPES..............................................................................
52
3.2 PRODUCTION AND CHARACTERIZATION OF WILDTYPE HIV-VLPS
(WT-VLPS)...............53
3.3 INTRASTRUCTURAL HELP BY GAG IM
MUNOGENS............................................................54
3.3.1 DOSE-FINDING
EXPERIMENT............................................................................................54
3.3.2 GAG-SPECIFIC T CELL RESPONSES AFTER
PRIMING...........................................................59
3.3.3 MODULATION OF THE HIV ENV SPECIFIC ANTIBODY RESPONSE BY AN
ADJUVANTED HIV
GAG-PROTEIN
IMMUNIZATION........................................................................................
62
3.4 HETEROLOGOUS INTRASTRUCTURAL HELP BASED ON
TETANUS-TOXOID.......................66
3.4.1 GENERATION OF VLP CONTAINING FRAGMENT C OF
TETANUS-TOXOID...............................67
3.4.2 IMPACT OF TETANOL PRIMING ON ENV-SPECIFIC IGG RESPONSES TO VLP
VACCINES
CONTAINING
TT-FRC.......................................................................................................67
3.4.3 IMPACT OF TETANOL IMMUNIZATION ON ENV-SPECIFIC IGG RESPONSES TO
VLP
VACCINES CONTAINING TT-P2D1 T HELPER CELL EPITOPES
...........................................
68
3.4.3.1 GENERATION AND CHARACTERIZATION OF VLPS INCORPORATING P2D1
.......................
69
3.4.3.2 HUMORAL IMMUNE RESPONSE INDUCED BY THE LICENSED TETANOL VACCINE
..........
70
3.4.3.3 ANALYSIS OF TETANUS-TOXOID BASED INTRASTRUCTURAL HELP
.......................................
72
3.4.4 TETANUS-SPECIFIC T CELL
RESPONSES.........................................................................
73
3.4.5 IMPACT OF ISH ON THE FUNCTIONALITY OF ENV-SPECIFIC ANTIBODIES
..................
75
3.4.5.1 HIV-1 ENV ANTIBODY AVIDITY
....................................................................................
75
3.4.5.2 FEY EFFECTOR
FUNCTIONALITY.........................................................................................
77
3.4.5.3 DURABILITY OF INDUCED ENV-SPECIFIC
ANTIBODIES.....................................................78
3.5 ANALYSIS OF HBSAG BASED INTRASTRUCTURAL HELP
...................................................
79
3.5.1 HBSAG T HELPER EPITOPE SCREENING
.........................................................................
80
3.5.2 HB-GAG-VLPS PRODUCTION AND CHARACTERIZATION
....................................................
84
3.5.3 HUMORAL RESPONSE AFTER PRIMING IMMUNIZATION
.....................................................
85
3.5.4 IMPACT OF HBVAXPRO PRIMING ON ENV-SPECIFIC RESPONSE AFTER
HB-GAG-VLP
BOOSTER
IMMUNIZATION................................................................................................
86
3.5.5 HBSAG-SPECIFIC T CELL RESPONSES, DNA VERSUS LICENSED VACCINE
.......................
89
3.5.6 IMPACT OF ISH ON THE FUNCTIONALITY OF ENV-SPECIFIC
ANTIBODIES....................91
4
DISCUSSION........................................................................................................................93
4.1 ADJUVANTED PROTEIN VACCINE INTRASTRUCTURAL HELP
.......................................................
93
4.2 TETANUS-TOXOID INTRASTRUCTURAL
HELP..............................................................................
96
4.3 HBSAG INTRASTRUCTURAL
HELP..........................................................................................100
5
BIBLIOGRAPHY..................................................................................................................
105
|
| any_adam_object | 1 |
| author | Elsayed, Hassan |
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| ctrlnum | (OCoLC)1135622762 (DE-599)DNB1176654942 |
| discipline | Medizin |
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| language | English |
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| spelling | Elsayed, Hassan Verfasser aut Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten vorgelegt von Hassan Elsayed aus Sharkia, Ägypten Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten Erlangen ; Nürnberg 2018 viii, 116 Seiten Illustrationen, Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Friedrich-Alexander-Universität Erlangen-Nürnberg 2018 Immunreaktion (DE-588)4133841-8 gnd rswk-swf Virus-like particles (DE-588)1078933014 gnd rswk-swf HIV (DE-588)4200792-6 gnd rswk-swf T-Lymphozyt (DE-588)4127387-4 gnd rswk-swf Impfstoff (DE-588)4026655-2 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content HIV (DE-588)4200792-6 s T-Lymphozyt (DE-588)4127387-4 s Virus-like particles (DE-588)1078933014 s Immunreaktion (DE-588)4133841-8 s Impfstoff (DE-588)4026655-2 s DE-604 B:DE-101 application/pdf http://d-nb.info/1176654942/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030833348&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Elsayed, Hassan Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten Immunreaktion (DE-588)4133841-8 gnd Virus-like particles (DE-588)1078933014 gnd HIV (DE-588)4200792-6 gnd T-Lymphozyt (DE-588)4127387-4 gnd Impfstoff (DE-588)4026655-2 gnd |
| subject_GND | (DE-588)4133841-8 (DE-588)1078933014 (DE-588)4200792-6 (DE-588)4127387-4 (DE-588)4026655-2 (DE-588)4113937-9 |
| title | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| title_alt | Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| title_auth | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| title_exact_search | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| title_full | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten vorgelegt von Hassan Elsayed aus Sharkia, Ägypten |
| title_fullStr | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten vorgelegt von Hassan Elsayed aus Sharkia, Ägypten |
| title_full_unstemmed | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten vorgelegt von Hassan Elsayed aus Sharkia, Ägypten |
| title_short | Harnessing T helper cell responses induced by licensed vaccines for improving virus-like particle based vaccination against HIV |
| title_sort | harnessing t helper cell responses induced by licensed vaccines for improving virus like particle based vaccination against hiv verbesserung partikel basierter hiv impfstoffe durch rekrutierung bereits bestehender t helferzellantworten |
| title_sub | = Verbesserung Partikel-basierter HIV Impfstoffe durch Rekrutierung bereits bestehender T-Helferzellantworten |
| topic | Immunreaktion (DE-588)4133841-8 gnd Virus-like particles (DE-588)1078933014 gnd HIV (DE-588)4200792-6 gnd T-Lymphozyt (DE-588)4127387-4 gnd Impfstoff (DE-588)4026655-2 gnd |
| topic_facet | Immunreaktion Virus-like particles HIV T-Lymphozyt Impfstoff Hochschulschrift |
| url | http://d-nb.info/1176654942/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=030833348&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT elsayedhassan harnessingthelpercellresponsesinducedbylicensedvaccinesforimprovingviruslikeparticlebasedvaccinationagainsthivverbesserungpartikelbasierterhivimpfstoffedurchrekrutierungbereitsbestehenderthelferzellantworten AT elsayedhassan verbesserungpartikelbasierterhivimpfstoffedurchrekrutierungbereitsbestehenderthelferzellantworten |
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