Molecular Studies of TNF-alpha and IL-6 Gene Expression Upon FcgammaRI Activation

Boyd, Jamie S (2001) Molecular Studies of TNF-alpha and IL-6 Gene Expression Upon FcgammaRI Activation. MSc(R) thesis, University of Glasgow.

Full text available as:
[img]
Preview
PDF
Download (11MB) | Preview

Abstract

The high affinity Fc receptor for IgG, FcgammaRI, is expressed constitutively on the surface of macrophages and monocytes and plays a crucial role in the co-ordination of the humoral and cellular arms of the human immune system. The activation of the receptor by aggregation at the cell surface leads to recruitment of accessory signalling molecules the activation of a wide range of responses including antibody-directed cell cytotoxicity. However, the nature of the response is dependent on the differentiation state of the cell. It has been previously reported that in the monocyte-like U937 cell line when the cell is differentiated to a more macrophage-like phenotype a molecular switch causes an absolute change in the signal cascade that FcgammaRI initiates. However the effects of this switch on levels of gene expression in the nucleus have not been investigated. The work presented here details the development of two complementary assay systems to measure the levels of expression of the cytokines TNF-alpha and IL-6 in response to the activation of FcgammaRI in monocytes and macrophages. A competitive RT-PCR assay was developed to measure levels of mRNA in the cell and a reporter gene assay was developed to study in more detail the promoter sequences of the two genes. Both assays were shown to work in principle using the TNF-alpha mRNA and gene promoter, and transfection experiments utilising the TNF-alpha promoter coupled to a reporter gene showed a change in expression in response to induced cell differentiation. Although the work proceeded no further than this stage, it provided two valuable tools for further examination of the novel mechanism FcgammaRI employs to control gene expression.

Item Type: Thesis (MSc(R))
Qualification Level: Masters
Additional Information: Adviser: Janet M Allen
Keywords: Medicine, Genetics
Date of Award: 2001
Depositing User: Enlighten Team
Unique ID: glathesis:2001-75723
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 19 Dec 2019 09:15
Last Modified: 19 Dec 2019 09:15
URI: http://theses.gla.ac.uk/id/eprint/75723

Actions (login required)

View Item View Item

Downloads

Downloads per month over past year