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oai:uvadoc.uva.es:10324/551782022-09-19T19:00:17Zcom_10324_1134com_10324_931com_10324_894col_10324_1213

Treatment with bortezomib of human CD4+ T cells preserves natural regulatory T cells and allows the emergence of a distinct suppressor T-cell population Blanco Durango, Belén Perez Simon, José Antonio Sánchez Abarca, Luis Ignacio Caballero Velazquez, Teresa Gutierrez Cossio, Silvia Hernández Campo, Pilar Diez Campelo, María Herrero Sánchez, María del Carmen Rodríguez Serrano, Concepción Santamaría Quesada, Carlos Sánchez Guijo, Fermín Cañizo Fernández-Roldán, Consuelo del San Miguel Izquierdo, Jesús F. Cell Biology Graft versus host disease Cell transplantation Trasplante de células Medical microbiology Pharmacology Producción Científica Background In vitro depletion of alloreactive T cells using the proteasome inhibitor bortezomib is a promising approach to prevent graft-versus-host disease after allogeneic stem cell transplantation. We have previously described the ability of bortezomib to selectively eliminate alloreactive T cells in a mixed leukocyte culture, preserving non-activated T cells. Due to the role of regulatory T cells in the control of graft versus host disease, in the current manuscript we have analyzed the effect of bortezomib in regulatory T cells.Design and Methods Conventional or regulatory CD4+ T cells were isolated with immunomagnetic microbeads based on the expression of CD4 and CD25. The effect of bortezomib on T-cell viability was analyzed by flow cytometry using 7-amino-actinomycin D staining. To investigate the possibility of obtaining an enriched regulatory T-cell population in vitro with the use of bortezomib, CD4+ T cells were cultured during four weeks in the presence of anti-CD3 and anti-CD28 antibodies, IL-2 and bortezomib. The phenotype of these long-term cultured cells was studied, analyzing the expression of CD25, CD127 and FOXP3 by flow cytometry, and mRNA levels were determined by RT-PCR. Their suppressive capacity was assessed in co-culture experiments, analyzing proliferation and IFN-γ and CD40L expression of stimulated responder T cells by flow cytometry.Results We observed that naturally occurring CD4+CD25+ regulatory T cells are resistant to the pro-apoptotic effect of bortezomib. Furthermore, we found that long-term culture of CD4+ T cells in the presence of bortezomib promotes the emergence of a regulatory T-cell population that significantly inhibits proliferation, IFN-γ production and CD40L expression among stimulated effector T cells.Conclusions These results reinforce the proposal of using bortezomib in the prevention of graft versus host disease and, moreover, in the generation of regulatory T-cell populations, that could be used in the treatment of multiple T-cell mediated diseases. 2022-09-19T08:26:32Z 2022-09-19T08:26:32Z 2009 info:eu-repo/semantics/article Haematologica, 2009, Vol. 94, Nº. 7, págs. 975-983 0390-6078 https://uvadoc.uva.es/handle/10324/55178 10.3324/haematol.2008.005017 975 7 983 Haematologica 94 1592-8721 eng https://haematologica.org/article/view/5306 info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ © Ferrata Storti Foundation Attribution-NonCommercial-NoDerivatives 4.0 Internacional Ferrata Storti Foundation SI