Characterization of a Cytomegalovirus-Specific T Lymphocyte Product Obtained Through a Rapid and Scalable Production Process for Use in Adoptive Immunotherapy
Grau-Vorster, Marta (Hospital Universitari Vall d'Hebron. Institut de Recerca)
López-Montañés, María (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Cantó Puig, Ester (Hospital Universitari Vall d'Hebron. Institut de Recerca)
Vives Armengol, Joaquim (Universitat Autònoma de Barcelona. Departament de Medicina)
Oliver-Vila, Irene (Cell Therapy Service, Banc de Sang i Teixits)
Barba, Pere (Hospital Universitari Vall d'Hebron)
Querol, Sergi (Cell Therapy Service, Banc de Sang i Teixits)
Rudilla, Francesc (Hospital Universitari Vall d'Hebron. Institut de Recerca)

Date:	2020
Abstract:	Immunosuppressed patients are susceptible to virus reactivation or de novo infection. Adoptive immunotherapy, based on virus-specific T lymphocytes (VST), can prevent or treat viral diseases. However, donor availability, HLA-compatibility restrictions, high costs, and time required for the production of personalized medicines constitute considerable limitations to this treatment. Ex vivo rapid and large-scale expansion of VST, compliant with current good manufacturing practice (cGMP) standards, with an associated cell donor registry would overcome these limitations. This study aimed to characterize a VST product obtained through an expansion protocol transferable to cGMP standards. Antigenic stimulus consisted of cytomegalovirus (CMV) pp65 peptide pool-pulsed autologous dendritic cells (DCs) derived from monocytes. G-Rex technology, cytokines IL-2, IL-7, and IL-15, and anti-CD3 and anti-CD28 antibodies were used for culture. At day 14 of cell culture, the final product was characterized regarding T cell subsets, specificity, and functionality. The final product, comprised mainly CD4 + and CD8 + T lymphocytes (49. 2 ± 24. 7 and 42. 3 ± 25. 2, respectively). The culture conditions made it possible to achieve at least a 98. 89-fold increase in pp65-specific CD3 + IFN-γ + cells. These cells were specific, as pp65-specific cytotoxicity was demonstrated. Additionally, in complete HLA mismatch and without the presence of pp65, alloreactivity resulted in <5% cell lysis. In conclusion, a cGMP scalable process for the generation of a large number of doses of CMV-specific cytotoxic T cells was successfully performed.
Grants:	Instituto de Salud Carlos III I FIS16/01433
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Virus specific T lymphocytes (VST) ; Antigen presenting cells (APC) ; Adoptive immunotherapy ; Specificity ; Cytotoxicity ; Peripheral blood mononuclear cells (PBMC) ; Alloreactivity
Published in:	Frontiers in immunology, Vol. 11 (february 2020) , ISSN 1664-3224

DOI: 10.3389/fimmu.2020.00271
PMID: 32161589

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