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Title: Cellular gene expression profiles of human macrophages exposed to Chlamydia pneumoniae and treated with low density lipoprotein
Keywords: Atherosclerosis, Chlamydia Pneumoniae, Low density lipoprotein, Differential Display RT-PCR, Real Time PCR, Foam cells
Issue Date: 3-Feb-2005
Citation: LIM CHIN TIONG, WILLIAM (2005-02-03). Cellular gene expression profiles of human macrophages exposed to Chlamydia pneumoniae and treated with low density lipoprotein. ScholarBank@NUS Repository.
Abstract: Chlamydia pneumoniae is an obligate intracellular bacterium which has been reported to be associated with atherosclerosis. The presence of Chlamydia pneumoniae in the diseased state as opposed to the normal state signified its possible association. Differential display (DD)-RT-PCR was the method employed to analyze mRNA derived from U937 macrophages and/or treatment with low density lipoprotein. Low density lipoprotein is another key factor being taken into consideration as foam cells are formed when macrophages take up oxidized low density lipoproteins. Five study models have been designated in our study, modeled after possible formation or events leading to atherosclerosis in the presence of Chlamydia pneumoniae and low density lipoproteins. We have isolated, sequenced and identified 235 cDNA fragments that shows different expression profiles within the study models as compared to the controls at the various time points, 24, 72 and 96 hours respectively. 190 were found to exhibit differential expression whereas 45 displayed unaltered expression profiles. 32 differentially expressed mRNAs showed no similarity hits whereas 158 of the differentially expressed transcripts matched known genes, which encodes for components involved in the ubiquitination (nuc2, CDC27, CTSH and HSPC150), cell proliferation (BCL2A1, CDC2, I?-NAC, CDC42), immune response (BLAME, C1QBP, PPP2RB,GPR6), structural (TUBE1, FLOT1, ACTG1 and TPM1) and components of the protein translation and modifications (EEF1A1, IF2). Real time PCR and semi quantitative RT-PCR is used to authenticate the altered expression profiles of the 29 selected genes of interest observed within the models. The data obtained justified the approach of using mRNA differential display in our study of Chlamydia pneumoniae and low density lipoprotein towards the development and progression of the atherosclerosis of which it is known to be caused by inflammation.
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