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Novel approaches for the isolation and expansion of cytomegalovirus-specific T cells for adoptive immunotherapy

Samuel, ER; (2013) Novel approaches for the isolation and expansion of cytomegalovirus-specific T cells for adoptive immunotherapy. Doctoral thesis , UCL (University College London). Green open access

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Abstract

CMV infections post hematopoietic stem cell transplantation (HSCT) can be effectively controlled through the adoptive transfer of donor derived CMV-specific T cells (CMV-T). Current strategies involve a second leukapheresis collection from the original donor to manufacture CMV-T, which is undesirable in the related donor setting and often not possible in unrelated donor transplants. To overcome these limitations I have investigated using a small aliquot of the original granulocyte-colony stimulating factor (G-CSF) mobilised HSCT graft to manufacture CMV-T. I have explored the T cell response to CMVpp65 peptide stimulation in G-CSF mobilised PBMC and subsequently examined the identification and isolation of CMV-T based on the activation markers CD154 and CD25. CD25+ enriched CMV-T from G-CSF mobilised PBMC were shown to contain a higher proportion of FoxP3 expressing cells than non-mobilised PBMC and also showed superior suppression of T cell proliferation. The successful expansion of CMV-T enriched through CD154 created a product which contained CD4+ and CD8+ cells, demonstrated a high specificity for CMV, secreted cytotoxic effector molecules and lysed CMVpp65 peptide loaded PHA blasts. These data provided the first published evidence that CMV-T can be effectively manufactured from G-CSF mobilised PBMC and that they share the same characteristics as CMV-T isolated in an identical manner from conventional non-mobilised PBMC. This provides a novel strategy for adoptive immunotherapy that abrogates the need for successive donation. The final part of the thesis explored the clinical scale-up of CD154 based enrichment of CMV-T from a G-CSF mobilised donor to assess the feasibility of manufacturing a viable therapeutic cell therapy. This has involved developing a GMP compliant, clinical scale system for the isolation of functional CD154+ CMV-T and has demonstrated the feasibility of translating this process into routine clinical practice, for the prophylactic and pre-emptive treatment of CMV reactivation following allogeneic HSCT.

Type: Thesis (Doctoral)
Title: Novel approaches for the isolation and expansion of cytomegalovirus-specific T cells for adoptive immunotherapy
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Third-party copyright material has been removed from this thesis.
UCL classification: UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Cancer Institute > Research Department of Haematology
URI: http://discovery.ucl.ac.uk/id/eprint/1421260
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