The function and regulation of single immunoglobulin IL-1-related receptor (SIGIRR) in kidney disease [Elektronische Ressource] / vorgelegt von Maciej Marcin Lech

Maciej Lech

Sujets

• Biologie
• Gesundheit

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 7
• Langue: English
• Poids de l'ouvrage: 4 Mo

Extrait

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Aus der Medizinischen Poliklinik – Innenstadt
der Ludwig-Maximilians-Universität München
Komm.Direktor: Prof.Dr.med. Martin Reincke

The function and regulation of single
immunoglobulin IL-1-related receptor (SIGIRR)
in kidney disease


Dissertation
zum Erwerb des Doktorgrades der Humanbiologie
an der Medizinischen Fakultät der
Ludwig-Maximilians-Universität
zu München


vorgelegt von
Maciej Marcin Lech

2007 2


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Mit Genehmigung der Medizinischen Fakultät der Ludwig-Maximilians-Universität,
München




Berichterstatter: Priv.Doz.Dr. H.-J. Anders
2. Berichterstatter: Prof.Dr. W.Samtleben

Mitberichterstatter: Prof.Dr. W. Zimmermann
Prof.Dr. V. Jansson


Dekan: Prof.Dr.med. D. Reinhardt

Tag der mündlichen Prüfung: 11.12.2007





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Acknowledgements

I would like to sincerely thank my supervisor, PD Dr. Hans Joachim Anders, for his trust in my skills,
support and guidance during my work in the laboratory. I am grateful his big interest in my project
and plenty of helpful discussions and for the scientific freedom I had.

My special thanks to Dr. Bruno Luckow, who has also been abundantly helpful; for all the
constructive discussion and help in solving scientific problems; and Prof. Detlef Schlöndorff for
providing the opportunities to discuss this research work and for his valuable suggestions.
I am grateful to all the Clinical Biochemistry Department lab members, for the atmosphere that they
have created. Thank you for all the help in my project and for all the fun that we had with each other.
Working with you was a pleasure.

I cannot end without thanking my friends, on whose constant encouragement and support I have
relied. This work would not have been possible without You. Thanks for the great vibes You have
been generating; for helping me deal with all the problems; for your love and trust in me and that I
could call this foreign city home.

Z całego serca dziękuję mojej rodzinie; dziękuję za Wasze wsparcie, miłość i wyrozumiałość.
W podzięce za Wasz trud i cięŜką pracę, ktŏre sprawiły, Ŝe osiągnąłem tak wiele, dedykuję Wam tę
pracę.

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LIST OF ABBREVIATIONS:

AP-1 – activator protein 1
APC – antigen presenting cell
ATF – activating transcription factor
BMDC – bone marrow dendritic cell
BSA – bovine serum albumin
CAM – cell adhesion molecule
cDNA – complementary DNA
DC – dendritic cell
DEPC – diethylene pyrocarbonate
DMEM – Dulbecco’s modified Eagle’s medium
DNA – deoxyribonucleic acid
DNTPs – deoxynucleotide triphosphates
ECD – extracellular domain
ECM – extracellular matrix
EGF – epidermal growth factor
ELISA – enzyme-linked immuno sorbent assay
FAS (APT1) – apoptosis antigen 1
FCS – fetal calf serum
FITC – fluorescein isothiocyanate
Flt3L – FMS-like tyrosine kinase 3 ligand
GMCSF – granulocyte-macrophage colony-stimulating factor
HEK – human epithelial kidney cells
HMGBP1 – high mobility group box protein 1
IC – immune complex
ICD – intracellular domain
IFN – interferon
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Ig – immunoglobulin
IL-1R – interleukin 1 receptor
IRAK – interleukin-1 receptor associated kinase
IRF – interferon regulatory factor
ITS – insulin/transferrine/selenium
JNK – C-jun N-terminal protein kinase
KC – keratinocyte-derived chemokine
LB – luria-bertani broth
LBP – LPS binding protein
LPS – lipopolysacharide
MCP-1 (CCL2) – monocyte chemoattractant protein-1
MD-2 – myeloid differentiation protein 2
MDA5 – melanoma differentiation associated protein 5
MHC – major histocompatibility complex
MIP-2 (CXCL2) – macrpphage inflammatory protein 2
MODS – multiple organ disfunction syndrome
MOF – multiple organ failure
MyD88 – myeloid differentiation protein 88
NADH – nicotinamidadenindinukleotid
NF- B – nuclear factor B
NLS – nuclear localization sequence
NOD – nucleotide-binding oligomerization domain
PAMPs – pathogen-associated molecular patterns
PBS – phosphate-buffered saline
PCR – polimerase chain reaction
PE – phycoerythrin
PGE-1 – prostaglandin 1
PI3K – phosphoinositide 3-kinase

kk9
PRR – pattern recognition receptors
PS – penicillin/streptomycin
RIG – retinoic acid-inducible gen 1
RNA – ribonucleic acid
RT – reverse transcriptase
SARM – sterile alpha and HEAT/armadillo motif protein
SIGIRR – single immunoglobulin IL-1-related receptor
SLE – systemic lupus erythematosus
SNP – single nucleotide polymorphism
snRNP – small nuclear ribonucleoproteins
SOCS-1 – suppressor of cytokine signaling 1
ss/ds – single-, double-stranded
TBS – tris buffered saline
TIR8 – toll interleukin-1 receptor 8
TIZ – TRAF6-inhibitory zinc finger protein
TLR – toll-like receptor
TMB – 3,3’,5,5’-tetramethylbenzidine
TNF – tumor necrosis factor
TRAF – TNF receptor associated factor
TRAM – TRIF-related adaptor molecule
TRIF – TNF receptor-inhibitory factor
U - unit
ZCCHC11 – Zinc finger CCHC domain containing protein 11
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TABLE OF CONTENTS

1. INTRODUCTION

1.1. Innate and adaptive immune system
1.2. TLR/IL-1R superfamily signaling
1.3. Regulation of TLR/IL-1R superfamily signaling
1.4. Structure and function of SIGIRR
1.5. Aim of the research project

2. MATERIAL & METHODS

2.1. Molecular biology methods
2.1.1. Cloning of human and murine SIGIRR
2.1.2. Electrophoresis of DNA
2.1.3. Ligation of DNA fragments
2.1.4. Generation of competent cells for electro-transformation
2.1.5. Transformation of bacterial cells by electroporation
2.1.6. Small scale plasmid preparation – mini prep
2.1.7. Analysis of DNA with restriction enzymes
2.1.8. RNA isolation from cultured cells
2.1.9. Isolation of RNA from tissues
2.1.10. cDNA synthesis and the real-time PCR analysis
2.1.11. Northern blot
2.2. Cell culture methods and cytokine detection
2.2.1. Cell lines
2.2.2. Calcium phosphate transfection
2.2.3. Reporter gene analysis
2.2.4. Primary cell culture – tubular epithelial cells
2.2.5. Primary cell culture – mesangial cells