eprintid: 1419041 rev_number: 39 eprint_status: archive userid: 608 dir: disk0/01/41/90/41 datestamp: 2014-02-11 19:43:59 lastmod: 2020-02-13 06:12:10 status_changed: 2014-02-11 19:43:59 type: article metadata_visibility: show item_issues_count: 0 creators_name: Samuel, ER creators_name: Beloki, L creators_name: Newton, K creators_name: Mackinnon, S creators_name: Lowdell, MW title: Isolation of Highly Suppressive CD25+FoxP3+ T Regulatory Cells from G-CSF-Mobilized Donors with Retention of Cytotoxic Anti-Viral CTLs: Application for Multi-Functional Immunotherapy Post Stem Cell Transplantation. ispublished: pub divisions: UCL divisions: A01 divisions: B02 divisions: C10 divisions: D19 divisions: G98 note: ©� 2014 Samuel et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. PMCID: PMC3895016 abstract: Previous studies have demonstrated the effective control of cytomegalovirus (CMV) infections post haematopoietic stem cell transplant through the adoptive transfer of donor derived CMV-specific T cells (CMV-T). Strategies for manufacturing CMV immunotherapies has involved a second leukapheresis or blood draw from the donor, which in the unrelated donor setting is not always possible. We have investigated the feasibility of using an aliquot of the original G-CSF-mobilized graft as a starting material for manufacture of CMV-T and examined the activation marker CD25 as a targeted approach for identification and isolation following CMVpp65 peptide stimulation. CD25+ cells isolated from G-CSF-mobilized apheresis revealed a significant increase in the proportion of FoxP3 expression when compared with conventional non-mobilized CD25+ cells and showed a superior suppressive capacity in a T cell proliferation assay, demonstrating the emergence of a population of Tregs not present in non-mobilized apheresis collections. The expansion of CD25+ CMV-T in short-term culture resulted in a mixed population of CD4+ and CD8+ T cells with CMV-specificity that secreted cytotoxic effector molecules and lysed CMVpp65 peptide-loaded phytohaemagglutinin-stimulated blasts. Furthermore CD25 expanded cells retained their suppressive capacity but did not maintain FoxP3 expression or secrete IL-10. In summary our data indicates that CD25 enrichment post CMV stimulation in G-CSF-mobilized PBMCs results in the simultaneous generation of both a functional population of anti-viral T cells and Tregs thus illustrating a potential single therapeutic strategy for the treatment of both GvHD and CMV reactivation following allogeneic haematopoietic stem cell transplantation. The use of G-CSF-mobilized cells as a starting material for cell therapy manufacture represents a feasible approach to alleviating the many problems incurred with successive donations and procurement of cells from unrelated donors. This approach may therefore simplify the clinical application of adoptive immunotherapy and broaden the approach for manufacturing multi-functional T cells. date: 2014-01-17 official_url: http://dx.doi.org/10.1371/journal.pone.0085911 vfaculties: VGHCSCI oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green article_type_text: Journal Article verified: verified_manual elements_source: PubMed elements_id: 926334 doi: 10.1371/journal.pone.0085911 pii: PONE-D-13-34036 lyricists_name: Lowdell, Mark lyricists_name: Mackinnon, Stephen lyricists_name: Samuel, Edward lyricists_id: MWLOW91 lyricists_id: SMACK49 lyricists_id: ESAMU32 full_text_status: public publication: PLoS One volume: 9 number: 1 article_number: e85911 event_location: United States citation: Samuel, ER; Beloki, L; Newton, K; Mackinnon, S; Lowdell, MW; (2014) Isolation of Highly Suppressive CD25+FoxP3+ T Regulatory Cells from G-CSF-Mobilized Donors with Retention of Cytotoxic Anti-Viral CTLs: Application for Multi-Functional Immunotherapy Post Stem Cell Transplantation. PLoS One , 9 (1) , Article e85911. 10.1371/journal.pone.0085911 <https://doi.org/10.1371/journal.pone.0085911>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/1419041/1/journal.pone.0085911.pdf