Molluscum contagiosum virus (MCV) is a significant human pathogen causing benign tumours in the human skin. Molluscum contagiosum (MC) infection is most common in children, young adults and immunodeficient individuals. MCV replicates well in the human skin in vivo, but conditions that support MCV replication in vitro are unknown. The lack of in vitro cell culture system has significantly limited progress in MCV research. The aims of this study were, (i) To develop a reporter assay for the sensitive detection and quantitation of MCV infections in vitro, (ii) To express suitable MCV virion surface antigens to develop a sensitive MCV ELISA assay, and (iii) To raise and characterize monoclonal antibodies (mAbs) against these antigens for the detection of MCV in infected human skin. All goals were achieved. A reporter assay based on simultaneous transfection of luciferase/EGFP reporter plasmids and infection with live MCV worked well and was used to test the infectivity of MCV in several human and animal cells. A novel C-terminal MC084 ELISA was established and used to determine MCV seroprevalence in two European populations. mAbs against MC084 and MC133 were raised and partially characterized. Interesting findings were that MCV not only infects human keratinocytes, but also a wide range of animal and human cells in vitro, which, however, do not support replication. Our MCV ELISA gives seroprevalences in UK and German general populations comparable with previous Australian and Japanese studies (<40 years of age). Surprisingly, in older age groups (vaccinated against smallpox), a significantly higher MCV seroprevalence was observed. This was not due to crossreactivity with VACV. We propose this increase may be due to childhood MCV antibodies being boosted by subsequent vaccination or vice versa. Finally, the mAbs raised are unique reagents and will be used in future work to test a hypothesis that MCV replicates in human epidermal stem cells.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:590361 |
| Date | January 2013 |
| Creators | Sherwani, Subuhi |
| Publisher | Cardiff University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://orca.cf.ac.uk/57429/ |
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