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Title: Pancreatic cancer as a target for adoptive T-cell immunotherapy
Author: Anderson, John Edward
ISNI:       0000 0001 3421 8727
Awarding Body: University of Manchester
Current Institution: University of Manchester
Date of Award: 2007
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The aim of this thesis was to investigate pancreatic cancer as a target for chimeric immune (CIR) based adoptive T ceil therapy and to irlcntiif\! presence of carcinoembryonic (CEA) and 5T4 antigen targets pancreatic tumour tissue. order to confirm the effectiveness of any it is important to identify a suitable model system. The immunotherapy mechanism on this thesis is based requires CIR specific recognition of the target antigens CEA or 5T4. Therefore two pancreatic and one gastric cancer cell expressing these were identified by immunofluorescent staining and Western Blot analysis. The general is numbers of T cells are required when targeting cancer tissue with adoptive immunotherapy. It is therefore necessary to be able to generate the of cells required a laboratory setting. different methods of T cell activation, using either anti-CD3s OKT3 or antibiotin T cell activation beads, were assessed comparing the rates of cell expansion, viability and cytokine production. To enable T cells to express CIR requires the genetic modification of T lymphocytes, also termed transduction. this thesis a retrovirus based on a murine leukaemia was used. As part the genetic modification process a retroviral clone expressing the 5T4 trans gene was produced. Pancreatic cancer patients' T cells were transduced two alternative transgenes . expressing a CIR specific for either the 5T4 or CEA antigen. levels of transduction efficiency achieved were assessed by staining and flow cytometer analysis. The specific cytotoxic function of the transduced cells was compared against that of untransduced T cells same patients by co-culturing the cells with the three Cancer lines previously shown to express CEA and 5T4 surface antigens. Cancer survival was assessed using a WST-1 cell proliferation assay. results confirmed specific cancer cell cytolysis was mediated by CIR T cells. After showing the adoptive immunotherapy model used was effective at reducing pancreatic and gastric cancer cell numbers vitro, it was important to identify the target antigens' expression true solid pancreatic cancer tissue specimens. Tissue was collected from pancreaticoduodenectomy that had been resected because of the suspicion cancer. Immunohistochemical staining identified the presence of CEA 5T4 antigens the pancreatic tumour tissue. of the findings of the in vitro model used this thesis and the expression of the target antigens indicated by immunohistochemical an~ true pancreatic cancer tissue, adoptive T cell immunotherapy have the potential, further development, to play a role the over survival outcome of pancreatic cancer patients.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available