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Title: An investigation of cell -type and stimulus specific expression patterns of inflammatory signal transduction mediators and their splice variants
Author: Salamaxani, Eleni
ISNI:       0000 0004 2743 2671
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2009
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The scope of research for this thesis is to see the biological relevance of cell type specific expression patterns of inflammation signalling genes by using computer based and experimental approaches. Our aim is to gain an enhanced understanding of the mechanisms that regulate inflammatory signalling. In order to do this we needed to define and identify the expression patterns of the individual inflammatory signalling components that are involved in Cardiovascular Disease (CVD). At the same time it was our endeavor to understand the relevance of isogenes and alternative splicing in inflammatory signalling process by studying the Mitogen Activated Protein Kinase (MAPK) signalling system. We tested genes JNKI and JNK2, and found that IL-lB, PMA and LPS regulate their expression in a cell type, time and stimulus specific manner. PMA seems to trigger JNKI expression in all tested cell lines, in a time dependent fashion. JNK2 shows a more restricted expression pattern depending on the cell line tested.IL-I Baffects both THP-l and endothelial cells where all 4 JNKI isoforms are regulated in a time dependent manner again. In addition, JNK2 is not expressed in these cells lines. LPS stimulated HeLa cells express both JNK 1 and JNK2 genes, once again regulated on a time dependent manner. The above findings providing the trigger and the foundation to try specify the biological relevance of JNK isoforms and see their differences. We created a JNKI negative cell line to check the JNK2 isoform expression. Thus we tried to clone the 4 different JNK2 isoforms and transfect the siJNK 1 HeLa cells. After stimulation of ILl b, to induce expression of endogenous IL-l b, we collected the samples after certain time points and compare the samples by Real-Time PCR.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available