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

Data:	2020
Resum:	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.
Ajuts:	Instituto de Salud Carlos III IFIS16/01433
Drets:	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.
Llengua:	Anglès
Document:	Article ; recerca ; Versió publicada
Matèria:	Virus specific T lymphocytes (VST) ; Antigen presenting cells (APC) ; Adoptive immunotherapy ; Specificity ; Cytotoxicity ; Peripheral blood mononuclear cells (PBMC) ; Alloreactivity
Publicat a:	Frontiers in immunology, Vol. 11 (february 2020) , ISSN 1664-3224

DOI: 10.3389/fimmu.2020.00271
PMID: 32161589

11 p, 1.6 MB

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