Combination of Epstein-Barr virus nuclear antigen 1, 3 and lytic antigen BZLF1 peptide pools allows fast and efficient stimulation of Epstein-Barr virus-specific T cells for adoptive immunotherapy.
Fiche publication
Date publication
janvier 2014
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BENSOUSSAN Danièle, Pr DE CARVALHO BITTENCOURT Marcelo, Dr JEULIN Hélène
Tous les auteurs :
Wang Y, Aissi-Rothe L, Virion JM, De Carvalho Bittencourt M, Ulas N, Audonnet S, Salmon A, Clement L, Venard V, Jeulin H, Stoltz JF, Decot V, Bensoussan D
Lien Pubmed
Résumé
BACKGROUND: Epstein-Barr virus (EBV) infection is a major cause of morbidity following hematopoietic stem cell transplantation. EBV-infected B cells may not respond to rituximab treatment and may lead to a life-threatening post-transplantation lymphoproliferative disorder. Adoptive cellular immunotherapy using EBV-lymphoblastoid cell lines (LCL) as stimulating antigen has proved effective in restoring specific immunity. However, EBV presents several immunodominant antigens, and developing a swift and effective clinical-grade immunotherapy relies on the definition of a Good Manufacturing Practices (GMP) universal stimulating antigen. METHODS: Peripheral blood mononuclear cells (PBMCs) from six donors with a cellular immune response against EBV were immunoselected after stimulation with a new EBV antigen associated with an EBNA3 peptide pool. RESULTS: After immunoselection, a mean of 0.53 +/- 0.25 x 10(6) cells was recovered consisting of a mean of 24.77 +/- 18.01% CD4(+)-secreting interferon (IFN)-gamma and 51.42 +/- 26.92% CD8(+)-secreting IFN-gamma. The T memory stem cell sub-population was identified. EBV-specific T cells were expanded in vitro, and their ability to secrete IFN-gamma and to proliferate after re-stimulation with EBV antigen was confirmed. A specific lysis was observed against autologous target cells pulsed with EBV peptide pools (57.6 +/- 11.5%) and against autologous EBV-LCL (18.3 +/- 7.3%). A mean decrease of 94.7 +/- 3.3% in alloreactivity against third-party donor mononuclear cells with EBV-specific T cells was observed compared with PBMCs before selection. CONCLUSIONS: Our results show that a combination of peptide pools including EBNA3 is needed to generate EBV-specific T cells with good specific cytotoxicity and devoid of alloreactivity, but as yet GMP grade is not fully achieved.
Référence
Cytotherapy. 2014 Jan;16(1):122-34