title: Rapid generation of full clinical-grade human antiadenovirus cytotoxic T cells for adoptive immunotherapy.
abstract: Adenovirus (ADV) infections are one of the major causes of morbidity and mortality after hematopoietic stem cell transplantation, despite new antiviral treatment strategies. We describe here a complete clinical-grade generation of human anti-ADV cytotoxic T cells to propose an adoptive immunotherapy. Peripheral blood mononuclear cells (PBMC) from 7 healthy donors, known for their good cellular immunity against ADV, were stimulated for 6 hours with a synthetic peptide pool covering the ADV5 Hexon protein interferon-gamma (IFN-gamma) secreting cells were isolated on a clinical device. After immunoselection, a mean number of 1.01 +/- 0.84 x 10(6) total nucleated cells was obtained. The isolated ADV-specific T cells were mainly CD4+ (mean=56% +/- 20.8%, yield=51% +/- 32.4%) but also CD8+ (mean=42% +/- 27%, yield = 56% +/- 39.3%). Isolated T lymphocytes (CTL) were expanded to carry out functional tests. Ability of the expanded CTL to secrete IFN-gamma and to proliferate after restimulation with the ADV peptide pool was confirmed. A high cytotoxicity against autologous target cells loaded with ADV antigens was observed but not against nonloaded target cells. We observed a decrease of 1.27 log of the allogeneic reaction against non HLA identical healthy donor PBMC with CTL compared with the PBMC before selection. Clinical-grade generation of ADV-specific T cells was achieved with a synthetic antigen. This technology has the advantage of being fast, and is sufficiently reactive to be proposed for immunotherapy if antiviral treatment fails.
authors.affiliation: CHU de Nancy, Unité de Thérapie Cellulaire et Tissus, Vandoeuvre-Les-Nancy, France.
authors.forename: Lamia
authors.initials: L
authors.lastname: Aïssi-Rothé
authors.forename: Véronique
authors.initials: V
authors.lastname: Decot
authors.forename: Véronique
authors.initials: V
authors.lastname: Venard
authors.forename: Hélène
authors.initials: H
authors.lastname: Jeulin
authors.forename: Alexandra
authors.initials: A
authors.lastname: Salmon
authors.forename: Laurence
authors.initials: L
authors.lastname: Clement
authors.forename: Anne
authors.initials: A
authors.lastname: Kennel
authors.forename: Christine
authors.initials: C
authors.lastname: Mathieu
authors.forename: Jean Hugues
authors.initials: JH
authors.lastname: Dalle
authors.forename: Georg
authors.initials: G
authors.lastname: Rauser
authors.forename: Christophe
authors.initials: C
authors.lastname: Cambouris
authors.forename: Marcelo
authors.initials: M
authors.lastname: de Carvalho
authors.forename: Jean François
authors.initials: JF
authors.lastname: Stoltz
authors.forename: Pierre
authors.initials: P
authors.lastname: Bordigoni
authors.forename: Danièle
authors.initials: D
authors.lastname: Bensoussan
mesh_terms: D000256:Adenoviridae / Q000276:immunology*; D000257:Adenoviridae Infections / Q000276:immunology / Q000628:therapy; D015704:CD4 Antigens / Q000096:biosynthesis; D036022:Capsid Proteins / Q000138:chemical synthesis / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology / Q000378:metabolism
chemical_list: D015704:CD4 Antigens; D036022:Capsid Proteins; D010446:Peptide Fragments; C018315:hexon capsid protein, Adenovirus; D007371:Interferon-gamma