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The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling
Dissertation (MSc)--University of Pretoria, 2011.
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| Format: | Thesis |
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University of Pretoria
2013
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| _version_ | 1867613516421136384 |
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| access_status_str | Open Access |
| author2 | Cromarty, Allan Duncan |
| author_browse | Cromarty, Allan Duncan |
| author_facet | Cromarty, Allan Duncan |
| collection | Thesis |
| dc_rights_str_mv | © 2011, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc)--University of Pretoria, 2011. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/25753 |
| institution | University of Pretoria (South Africa) |
| last_indexed | 2026-06-10T12:37:23.532Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2013 |
| publishDateRange | 2013 |
| publishDateSort | 2013 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/25753 The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling Cromarty, Allan Duncan Siderius, M. prishantha101@yahoo.co.uk Subramoney, Preya Us28 Antibody Tumour Cytokine Interleukin 6 il-6 Signal transducer Activator of transcription stat3 Oncogene Ligand G protein-coupled receptors gpcr Human cytomegalovirus hcmv UCTD Dissertation (MSc)--University of Pretoria, 2011. Human cytomegalovirus (HCMV) is a ubiquitous virus of the herpes type that infects a high percentage of some populations. One of the most researched genes expressed by HCMV with close homology to human chemokine receptors is the US28 G protein-coupled receptor. Study design: This study was initiated to elucidate the intracellular signalling pathways of an inflammatory factor (IL-6) and an angiogenic factor (STAT3) triggered by the viral US28 oncogene and the presence of US28 in the HCMV viral particle. These pathways were observed by introducing the US28 gene into two human cell lines by infection with a HCMV strain that expresses the US28 gene (wild type), and two HCMV strains where the US28 gene was deleted (ÄUS28 and ÄUS28/UL33). Special attention was directed at the expression of IL-6 after promotion of the US28 gene and subsequent phosphorolation of STAT3. A new US28 antibody was validated and a method developed in an attempt to determine US28 on the viral particle. The following techniques were applied: Cell culture work, two mammalian cell lines were used, HFF’s and U373 MG. Virus stock titre determination to determine the multiplicity of infection. Protein quantitation to determine very small quantities of protein for Western blot analysis. ELISA for the quantitative determination of IL-6. Western blotting for phospho- STAT3 determination and validation of the US28 antibody. Immunocytochemistry was used for back titrations of virally infected cells. Immunofluorescence assay and use of confocal microscopic techniques was used for the location of the US28 gene in the virion and for tSTAT3 translocation to the nucleus. Conclusion: A clear increase in IL-6 secretion (495% ± 1%) was seen, and this was after only an hour in HCMV WT infected cells. From the increase in IL-6 secretion a subsequent increase in STAT3 phosphorylation was detected in the same samples. A clear link has been established between IL-6 and STAT3. A method to determine whether US28 was present in the HCMV viral particle was designed and preliminary results obtained. The results were inclusive. Pharmacology unrestricted 2013-09-06T23:57:56Z 2011-06-23 2013-09-06T23:57:56Z 2011-04-08 2011-06-23 2011-06-22 Dissertation Subramoney, P 2011, The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://hdl.handle.net/2263/25753 > E11/299/gm http://hdl.handle.net/2263/25753 http://upetd.up.ac.za/thesis/available/etd-06222011-170730/ © 2011, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | Us28 Antibody Tumour Cytokine Interleukin 6 il-6 Signal transducer Activator of transcription stat3 Oncogene Ligand G protein-coupled receptors gpcr Human cytomegalovirus hcmv UCTD The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title | The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title_full | The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title_fullStr | The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title_full_unstemmed | The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title_short | The role of human cytomegalovirus encoded viral G protein-coupled receptors in onco-modulatory signalling |
| title_sort | role of human cytomegalovirus encoded viral g protein coupled receptors in onco modulatory signalling |
| topic | Us28 Antibody Tumour Cytokine Interleukin 6 il-6 Signal transducer Activator of transcription stat3 Oncogene Ligand G protein-coupled receptors gpcr Human cytomegalovirus hcmv UCTD |
| url | http://hdl.handle.net/2263/25753 http://upetd.up.ac.za/thesis/available/etd-06222011-170730/ |