TLR dependant hepcidin expression in THP-1 cells in response to TLR4 and TLR1/2 ligands.
Cardiff Metropolitan University
MetadataShow full item record
Iron is an essential mineral in all forms of life and in regards to pathological conditions its highly sought after as the availability of iron to the invading pathogen determines the fate of the pathogen, this desire to obtain and retain iron is revealed by the many systems whose role it is to acquire and protect/store iron. These systems have been observed in life ranging from microbes to higher organisms and hepcidin, the peptide of interest in this study is one which is conserved thought out many distinct species . In organisms such as bacteria these systems manifest themselves for the most part as high-affinity iron uptake mechanisms such as; siderophore-based systems, heme acquisition systems, and transferrin/lactoferrin receptors and these systems have been evolved to directly compete with the hosts high affinity iron withholding systems e.g. Hepcidin . Hepcidin is the master regulator of iron throughout the body and its induction under normal conditions is controlled by systemic iron and iron associated regulatory proteins, yet in a pathological situation a variety of TLR’s and cytokines are utilized to up regulate hepcidin and down regulate the sole iron exporter ferroportin (Fpn-1) . It’s this system that is under investigation, particularly TLR4 and TLR1/2 dimer dependent hepcidin induction via ligands LPS and Pam3CSK4 respectively  in THP-1 monocytes, macrophages and dendritic cells. Ligands were used to stimulate cell types for different lengths of time, 6 & 24 hour, and then mRNA was extracted using TRIzol and cDNA of HAMP gene generated via reverse-transcriptase-PCR, this DNA was then amplified to levels in which semi-quantitative analysis is viable, this being agarose gel electrophoresis and densitometry. Results quality is largely substandard as negative controls show evidence of contamination and the gel electrophoresis showed intrinsic problems with method and materials used as shown by the non-specific bandings and blurred bands, although dendritic cell samples provided enough data to conclude that dendritic cells up-regulate hepcidin in response to TLR ligands, this is a previously unknown yet relatively predictable response and with this information it opens other avenues of research into the role of hepcidin during inflammation and infection especially those concerning the adaptive immune system due to the dendritic cell being considered as the bridge between the innate and adaptive immune system..
BSc Biomedical Science
Showing items related by title, author, subject and abstract.
Al-Lawati, Hiba Abdul Redha (Cardiff Metropolitan University, 2011)Linking immunity with iron can be well illustrated by the production of hepcidin as it is considered to be the master regulator of iron metabolism in mammals. Iron is an essential element for microbes and higher animals ...
Al Shuraiqi, Rajaa (Cardiff Metropolitan University, 2011)Hepcidin a hepatic peptide hormone mainly produces by the liver tissue, it is also produce in a small amount by other tissue including lung, heart and macrophages. Hepcidin plays a critical role in iron regulation and ...
Whida, Amal Masre (Cardiff Metropolitan University, 2009)Iron has an important role in the survival of nearly all microorganisms, therefore, one of the strategies used by the immune system in the fight against infection is to reduce iron availability. Thus, iron regulation is ...