Identification of gene markers for in vitro toxicity control testing of pertussis vaccines

Moncassin, Noela (2005) Identification of gene markers for in vitro toxicity control testing of pertussis vaccines. MSc(R) thesis, University of Glasgow.

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Abstract

Whole-cell (WCV) and acellular (ACV) pertussis vaccines play an essential role in immunization programs for the prevention of the disease pertussis (whooping cough). Pertussis toxin (PT), one of the Bordetella pertussis toxins, is a major component in both WCV and ACV. The histamine sensitisation test (HIST) is a toxicity test, in mice, used to assure the absence of significant residual PT activity in pertussis vaccines. HIST is a lethal test and large variations in test performance have been observed. The objective of this project was to investigate the gene expression of selected cell lines treated with PT and detoxified pertussis toxin (dPT) using microarray technology and, from the expression profiles obtained, to identify gene markers of the toxicological effects of PT which may form the base to develop an in vitro assay as an alternative to HIST. Two cell types were investigated: human umbilical vein endothelial cells (HUVEC) and bronchial epithelial cells (NL20). Based on morphological and cytotoxicity studies, the cells were treated with PT and dPT at a concentration of 2.5 mug/ml for 6h. The gene expression profiles obtained lead to the speculation that PT could play an important role in the induction of the cell-mediated immunity (up-regulation of: galectin 3, small inducible cytokine subfamily 20, Thy-1 cell surface antigen and CD63 genes) and that PT could also have an important role in the induction of vascular permeability (up-regulation of: platelet-derived growth factor, vascular endothelial growth factor c) and that the effect may take place at the brain level (up- regulation of: glial fibrillary acidic protein, chlorine channel 3, cholinergic receptor). RT PCR study needs further investigation, nevertheless, the ICAM 1 gene (specifically up regulated by PT + TNF-alpha treatment) may serve as a gene marker of PT toxicity. ln addition, data from a cell migration study with endothelial cells suggested that PT could be involved in angiogenesis. The phenotypic and genomic data presented in this study suggest that, in order to develop a replacement for the in vivo HIST control test, an in vitro permeability assay with monolayer endothelial cells may be worth investigating.

Item Type: Thesis (MSc(R))
Qualification Level: Masters
Additional Information: Advisers: Dorothy Xing; Yuan Zhao
Keywords: Pharmaceutical sciences, Genetics, Toxicology
Date of Award: 2005
Depositing User: Enlighten Team
Unique ID: glathesis:2005-71151
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 10 May 2019 10:49
Last Modified: 10 May 2019 10:49
URI: http://theses.gla.ac.uk/id/eprint/71151

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