Published online 8 June 2009
Haematologica, Vol 94, Issue 7, 975-983 doi:10.3324/haematol.2008.005017
Copyright © 2009 by Ferrata Storti Foundation
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Blanco et al. - Appendix
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via Web of Science (1)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Blanco, B.
Right arrow Articles by San Miguel, J. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Blanco, B.
Right arrow Articles by San Miguel, J. F.

Cell Therapy and Immunotherapy

Treatment with bortezomib of human CD4+ T cells preserves natural regulatory T cells and allows the emergence of a distinct suppressor T-cell population

Belén Blanco, José A. Pérez-Simón, Luis I. Sánchez-Abarca, Teresa Caballero-Velazquez, Silvia Gutierrez-Cossío, Pilar Hernández-Campo, María Díez-Campelo, Carmen Herrero-Sanchez, Concepción Rodriguez-Serrano, Carlos Santamaría, Fermín M. Sánchez-Guijo, Consuelo del Cañizo, Jesús F. San Miguel

Servicio de Hematología, Hospital Universitario de Salamanca and Centro de Investigación del Cáncer (CIC/CSIC) Salamanca, Centro en Red de Medicina Regenerativa y Terapia celular de Castilla y León, Spain

Correspondence: José A Pérez Simón, Servicio de Hematología y CIC Salamanca, Paseo de San Vicente, s/n 37007, Salamanca, Spain. E-mail:pesimo{at}usal.es

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-{gamma} 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-{gamma} 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.

Key words: bortezomib, regulatory T cells, graft-versus-host disease, prevention.