Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | 129 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |c Matthias Müllenbeck |
| 264 | 1 | |a Berlin |c 2013 | |
| 300 | |a 129 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
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| 502 | |a Berlin, Humboldt-Univ., Diss., 2013 | ||
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| 776 | 0 | 8 | |i Reproduziert als |a Müllenbeck, Matthias |t Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |d Ketsch bei Mannheim : Mikroform Dissertation, 2013 |h 2 Mikrofiches |w (DE-604)BV041075352 |
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Datensatz im Suchindex
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IMAGE 1
III. TABLE OF CONTENTS
I. ABSTRACT 3
II. ZUSAMMENFASSUNG 4
III. TABLE OF CONTENTS 5
IV. ABBREVIATIONS 8
1. INTRODUCTION 12
1.1 GENERAL FEATURES O F THE HUMAN I M M U N E S Y S T E M 1 2
1.1 GENERATION O F ANTIBODY-DIVERSITY A N D ANTIBODY-SPECIFICITY WITHIN
HUMORAL I M M U N E R E S P O N S E S 1 2
1.2 GENERATION AND MAINTENANCE O F HUMORAL IMMUNOLOGICAL M E M O R Y 1 5
1.3 HUMORAL IMMUNOLOGICAL M E M O R Y IN HUMAN P/INFECTIONS 1 7
1.4 P/-LIFE CYCLE AND MEROZOITE-SPECIFIC M O D E L ANTIGENS 2 0
2. OBJECTIVES 2 4
3. METHODS 25
3.1. IN VITRO GENERATION O F HUMAN MONOCLONAL ANTIBODIES BY IG-GENE
EXPRESSION VECTOR CLONING O F SINGLE-CELL SORTED M E M O R Y B CELLS 2 5
3.1.1 SAMPLE ACQUISITION AND PBMC ISOLATION 2 6
3.1.2 FLUORESCENCE-ACTIVATED CELL SORTING (FACS) 2 6
3.1.3 REVERSE TRANSCRIPTION - POLYMERASE CHAIN REACTION 2 8
3.1.4 CDNA-SYNTHESIS 2 8
3.1.5 SEMI-NESTED PCR FOR AMPLIFICATION O F HUMAN IG G E N E TRANSCRIPTS
2 9
3.1.6 HUMAN IG G E N E S E Q U E N C E ANALYSIS 3 1
3.1.7 DIRECTIONAL CLONING INTO IGGL EXPRESSION VECTORS 3 2
3.1.8 PREPARATION O F EUKARYOTIC EXPRESSION VECTORS 3 2
3.1.9 RESTRICTION ENDONUCLEASE DIGESTION 3 2
3.1.10 LIGATION 3 4
3.1.11 PREPARATION AND TRANSFORMATION O F C O M P E T E N T BACTERIA 3 4
3.1.12 SCREENING O F BACTERIAL COLONIES BY PCR 3 5
3.1.13 PURIFICATION O F PLASMID DNA 3 6
3.1.14 RECOMBINANT ANTIBODY PRODUCTION 3 6
3.1.15 CELL CULTURE O F HEK293T CELLS 3 6
3.1.16 POLYETHYLENIMINE-MEDIATED DNA-TRANSFECTION O F HEK293T CELLS 3 7
3.1.17 ANTIBODY HARVEST AND PURIFICATION 3 7
3.2. ANTIBODY CHARACTERIZATION 3 8
3.2.1. ENZYME-LINKED I M M U N O S O R B A N T A S S A Y (ELISA) 3 8
3.2.1.1. MEASUREMENT OF IGG CONCENTRATIONS 38
3.2.1.2. REACTIVITY ELISA WITH PF ANTIGENS MSP3, GLURP, GMZ2 AND
SHORT-OVERLAPPING PEPTIDES THEREOF 39
3.2.1.3. POLYREACTIVITY ELISA WITH LPS, DNA, AND INSULIN 39
3.2.2. IMAGING O F P/-INFECTED RBCS (IRBCS) BY IMMUNOFLUORESCENT
MICROSCOPY 3 9 3.2.3. ANTIBODY IRBC-BINDING A S S E S S M E N T BY FACS
4 0
3.1.18 EVALUATION O F ANTIBODY-ANTIGEN INTERACTION B Y SURFACE PLASMON
RESONANCE * SCANS 4 1
3.2.5. IN VITRO P/-PARASITE CULTIVATION 4 1
3.2.6. A S S E S S M E N T O F ANTIBODY-MEDIATED NEUTRALIZATION O F P F
B Y A FIREFLY- LUCIFERASE EXPRESSING TRANSGENIC 3D7 PF- PARASITE LINE IN
VITRO 4 2
3.2.7. A S S E S S M E N T O F ANTIBODY-MEDIATED NEUTRALIZATION O F
P/-PARASITE LINES 3D7, HB3, DD2, AND IT2 BY SYBR GREEN STAINING IN VITRO
4 2
3.3. SECRETORY AND MEMBRANE IGG PCR 4 3
5
HTTP://D-NB.INFO/1030073848
IMAGE 2
TABLE O F CONTENTS
3.4. EX VIVO ANALYSIS O F S E R U M ANTIBODIES O F MALARIA I M M U N E
INDIVIDUALS B Y M A S S SPECTROMETRY 4 3
3.5. STATISTICAL ANALYSIS AND STOCHASTICAL SIMULATION 4 4
4. RESULTS 4 6
4.1. MALARIA I M M U N E AFRICAN INDIVIDUALS P O S S E S S HIGH TITERS O
F ANTI-P/IGG S E R U M ANTIBODIES WITH ^/-NEUTRALIZING ACTIVITY 4 6
4.2. MALARIA IMMUNE AFRICAN INDIVIDUALS S H O W HIGH FREQUENCY O F P /
RCACTIVE CM A N D ATM 4 9
4.3. SELECTION O F THREE INDIVIDUALS W I T H HIGH ^/-NEUTRALIZING SERUM
ANTIBODY TITERS AND HIGH FREQUENCIES O F P/-REACTIVE CM A N D ATM FOR IG
G E N E EXPRESSION VECTOR CLONING 5 1
4.4. IGG+ GMZ2-REACTIVE CM A N D ATM DIFFER IN THEIR LEVEL O F SOMATIC
HYPER MUTATIONS AND S H O W N O CLONAL RELATIONSHIP 5 3
4.5. /"/-REACTIVE ANTIBODIES O F CM AND ATM S H O W SIMILAR BINDING
CAPACITIES TO PF ANTIGENS 5 6
4.6. POLYREACTIVE ANTI-P/ANTIBODIES ARE ENRICHED IN T H E ATM
COMPARTMENT 5 9
4.7. CM AND ATM ANTIBODIES S H O W SIMILAR LEVELS O F P/-NEUTRALIXATION
FOR HB3 , DD2, AND IT2 P/-STRAIN 6 2
4.8. ATM EXPRESS ACTIVATION-ASSOCIATED SURFACE MOLECULES HLA-DR, CD24,
CD84, CD86 AND CD319 AND S H O W SIGNS O F ACTIVE PROLIFERATION 6 4
4.9. CM AND ATM ANTIBODIES ARE P R E S E N T IN T H E SERUM O F MALARIA
I M M U N E INDIVIDUALS 6 6
5. DISCUSSION 70
5.1. SELECTION O F THREE INDIVIDUALS WITH RELATIVE SUPERIOR P/-MALARIA
IMMUNITY 7 0 5.2. MONOMERIC FLUORESCENTLY-LABELED P/-ANTIGENS ALLOW
FACS-SORTING O F P/SPECIFIC M E M O R Y B CELLS 7 1
5.3. LACK O F SHARED CLONALLY EXPANDED B CELLS I N CM A N D ATM
INDICATES DIFFERENT ROUTES O F DEVELOPMENT IN P/-INFECTIONS 7 2
5.4. POLYREACTIVE ANTIBODIES ARE ENRICHED IN THE ANTI-P/ATM COMPARTMENT
7 3
5.5. ANTIBODIES O F CM AND ATM BROADLY NEUTRALIZE P/-MEROZOITES RBC
INVASION 7 6 5.6. ATM ARE ACTIVATED B CELLS THAT CONTRIBUTE TO THE
ANTI-P/SERUM ANTIBODY POOL 7 8 5.7. HUMORAL IMMUNOLOGICAL M E M O R Y TO
P/-MALARIA INFECTIONS IS GENERATED AND FUNCTIONAL 7 9
6. OUTLOOK 82
7. REFERENCES 8 3
8. SUPPLEMENTAL MATERIAL 9 4
8.1. ANTIBODIES 9 4
8.2. ANTIGENS 9 5
8.3. BACTERIA 9 6
8.4. BACTERIAL CULTURE MEDIA 9 6
8.5. BUFFERS AND SOLUTIONS 9 6
8.6. CELL LINES 1 0 2
8.7. CELL CULTURE MEDIUM 1 0 2
8.8. CHEMICALS 1 0 3
8.9. COMMERCIAL KITS 1 0 3
8.10. ENZYMES AND ADDITIVES.;. 1 0 3
8.11. EXPRESSION VECTORS 1 0 4
8.12. NUCLEOTIDES A N D NUCLEIC ACIDS 1 0 4
8.13. INSTRUMENTS A N D CONSUMABLES 1 0 5
8.14. SOFTWARE 1 0 8
8.15. WEB RESOURCES 1 0 9
6
IMAGE 3
9. SUPPLEMENTARY INFORMATION 110
10. SUPPLEMENTAL TABLES 115
11. APPENDIX 123
11.1. ACKNOWLEDGEMENTS 1 2 3
11.2. CURRICULUM VITAE 1 2 4
11.3. PUBLICATIONS AND POSTER 1 2 6
11.4. PATENTS 1 2 8
11.5. ETHICAL APPROVALS 1 2 8
11.6. STATEMENT O N OATH 1 2 9
7 |
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| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
| dewey-search | 610 |
| dewey-sort | 3610 |
| dewey-tens | 610 - Medicine and health |
| discipline | Medizin |
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| spelling | Müllenbeck, Matthias 1983- Verfasser (DE-588)1029987564 aut Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections Matthias Müllenbeck Berlin 2013 129 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Berlin, Humboldt-Univ., Diss., 2013 (DE-588)4113937-9 Hochschulschrift gnd-content Reproduziert als Müllenbeck, Matthias Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections Ketsch bei Mannheim : Mikroform Dissertation, 2013 2 Mikrofiches (DE-604)BV041075352 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025668338&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Müllenbeck, Matthias 1983- Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |
| subject_GND | (DE-588)4113937-9 |
| title | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |
| title_auth | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |
| title_exact_search | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |
| title_full | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections Matthias Müllenbeck |
| title_fullStr | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections Matthias Müllenbeck |
| title_full_unstemmed | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections Matthias Müllenbeck |
| title_short | Antibodies of classical- and atypical memory B cells neutralize human "Plasmodium falciparum" infections |
| title_sort | antibodies of classical and atypical memory b cells neutralize human plasmodium falciparum infections |
| topic_facet | Hochschulschrift |
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