Rapid generation of full clinical-grade human antiadenovirus cytotoxic T cells for adoptive immunotherapy.
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.
Aïssi-Rothé
Lamia
L
CHU de Nancy, Unité de Thérapie Cellulaire et Tissus, Vandoeuvre-Les-Nancy, France.
Decot
Véronique
V
Venard
Véronique
V
Jeulin
Hélène
H
Salmon
Alexandra
A
Clement
Laurence
L
Kennel
Anne
A
Mathieu
Christine
C
Dalle
Jean Hugues
JH
Rauser
Georg
G
Cambouris
Christophe
C
de Carvalho
Marcelo
M
Stoltz
Jean François
JF
Bordigoni
Pierre
P
Bensoussan
Danièle
D
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
D015704:CD4 Antigens; D036022:Capsid Proteins; D010446:Peptide Fragments; C018315:hexon capsid protein, Adenovirus; D007371:Interferon-gamma