Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy
Adoptive transfer of cytomegalovirus (CMV)-specific T cells has shown promising results in preventing pathological effects caused by opportunistic CMV infection in patients following allogeneic hematopoietic stem cell transplantation. The majority of studies have used steady-state leukapheresis for...
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| Format: | info:eu-repo/semantics/doctoralThesis |
| Language: | eng |
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Servicio de Publicaciones de la Universidad de Navarra
2015
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| Online Access: | https://hdl.handle.net/10171/38528 |
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| author | Beloki, L. (Lorea) Olavarría, E. (Eduardo) Ramirez, N. (Natalia) |
| author_facet | Beloki, L. (Lorea) Olavarría, E. (Eduardo) Ramirez, N. (Natalia) |
| author_sort | Beloki, L. (Lorea) |
| collection | DSpace |
| description | Adoptive transfer of cytomegalovirus (CMV)-specific T cells has shown promising results in preventing pathological effects caused by opportunistic CMV infection in patients following allogeneic hematopoietic stem cell transplantation. The majority of studies have used steady-state leukapheresis for CMV-reactive product manufacture, a collection obtained prior to or weeks after granulocyte-colony stimulating factor (G-CSF) mobilization, but the procurement of this additional sample is often not available in the unrelated donor setting. If the cellular product for adoptive immunotherapy could be generated from the same G-CSF mobilized collection, the problems associated with the additional harvest could be overcome. However, the tolerogenic effects induced by G-CSF on T cells could affect the anti-viral in vivo functionality of CMV-specific T cells obtained from G-CSF mobilized donor samples if they were infused into the patient.
In this thesis it has been investigated the use of G-CSF mobilized samples for the manufacture of CMV-specific T cells using two direct selection methods. First, CMV-specific T cells were isolated based on activation-dependent CD137 expression, which demonstrated a high specificity for CMV, the secretion of cytotoxic effector molecules, proliferation in response to the antigen and lysis of CMV-loaded target cells, showing that they shared the same characteristics as CMV-specific T cells obtained from non-mobilized cells. On the other hand, CMV-specific CD8+ T cells were isolated using MHC-multimers and showed that products obtained from G-CSF mobilized samples were able to respond to the antigen by expressing activation markers and secreting pro-inflammatory cytokines and lytic molecules, but the in vitro proliferative capacity was impaired. |
| format | info:eu-repo/semantics/doctoralThesis |
| id | oai:dadun.unav.edu:10171-38528 |
| institution | Universidad de Navarra |
| language | eng |
| publishDate | 2015 |
| publisher | Servicio de Publicaciones de la Universidad de Navarra |
| record_format | dspace |
| spelling | oai:dadun.unav.edu:10171-385282020-07-31T12:34:43Z Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy Beloki, L. (Lorea) Olavarría, E. (Eduardo) Ramirez, N. (Natalia) Materias Investigacion::Ciencias de la vida Transplante de órganos Inmunología Adoptive transfer of cytomegalovirus (CMV)-specific T cells has shown promising results in preventing pathological effects caused by opportunistic CMV infection in patients following allogeneic hematopoietic stem cell transplantation. The majority of studies have used steady-state leukapheresis for CMV-reactive product manufacture, a collection obtained prior to or weeks after granulocyte-colony stimulating factor (G-CSF) mobilization, but the procurement of this additional sample is often not available in the unrelated donor setting. If the cellular product for adoptive immunotherapy could be generated from the same G-CSF mobilized collection, the problems associated with the additional harvest could be overcome. However, the tolerogenic effects induced by G-CSF on T cells could affect the anti-viral in vivo functionality of CMV-specific T cells obtained from G-CSF mobilized donor samples if they were infused into the patient. In this thesis it has been investigated the use of G-CSF mobilized samples for the manufacture of CMV-specific T cells using two direct selection methods. First, CMV-specific T cells were isolated based on activation-dependent CD137 expression, which demonstrated a high specificity for CMV, the secretion of cytotoxic effector molecules, proliferation in response to the antigen and lysis of CMV-loaded target cells, showing that they shared the same characteristics as CMV-specific T cells obtained from non-mobilized cells. On the other hand, CMV-specific CD8+ T cells were isolated using MHC-multimers and showed that products obtained from G-CSF mobilized samples were able to respond to the antigen by expressing activation markers and secreting pro-inflammatory cytokines and lytic molecules, but the in vitro proliferative capacity was impaired. 2015-06-08T14:48:10Z 2015-06-08T14:48:10Z 2015 2014-12-13 info:eu-repo/semantics/doctoralThesis https://hdl.handle.net/10171/38528 eng info:eu-repo/semantics/openAccess application/pdf Servicio de Publicaciones de la Universidad de Navarra |
| spellingShingle | Materias Investigacion::Ciencias de la vida Transplante de órganos Inmunología Beloki, L. (Lorea) Olavarría, E. (Eduardo) Ramirez, N. (Natalia) Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title | Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title_full | Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title_fullStr | Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title_full_unstemmed | Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title_short | Evaluation and optimization of immunomagnetic methods for the purification and selection of CMV-specific cytotoxic T lymphocytes from G-CSF primed peripheral blood mononuclear cells for adoptive immunotherapy |
| title_sort | evaluation and optimization of immunomagnetic methods for the purification and selection of cmv-specific cytotoxic t lymphocytes from g-csf primed peripheral blood mononuclear cells for adoptive immunotherapy |
| topic | Materias Investigacion::Ciencias de la vida Transplante de órganos Inmunología |
| url | https://hdl.handle.net/10171/38528 |
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