CD28 costimulation and immunoaffinity-based selection efficiently generate primary gene-modified T cells for adoptive immunotherapy.
The introduction of an inducible suicide gene has been proposed as a strategy to exploit the antitumor reactivity of donor T cells after allogeneic hematopoietic stem cell transplantation but permit control of graft-versus-host disease. However, there are several obstacles to this approach that may impair the ability of T cells to function and survive in vivo. These include the requirement for in vitro activation or long-term culture to introduce the transgene and obtain therapeutic cell numbers, the toxicity of drug selection to enrich transduced cells, and the immunogenicity of the transgene-encoded products. Here we have developed a transduction and selection strategy for generating large numbers of polyclonal T cells transduced with a retroviral vector encoding the human low-affinity nerve growth factor receptor (LNGFR) for selection and a Fas-based suicide construct (LV'VFas). Ligation of CD28 in conjunction with a T-cell receptor signal permitted efficient transduction, substantially promoted T-cell growth, and contributed to the generation of gene-modified T cells that retained clonal diversity, functional properties, and a homing receptor profile similar to untransduced peripheral blood lymphocytes. Microbeads conjugated directly to antibody specific to LNGFR significantly improved the immunomagnetic selection of LV'VFas-modified T cells and assisted in scaling of the selection procedure to therapeutic cell numbers. Thus, these studies identified a strategy that requires only a brief ex vivo culture and does not use drug selection to obtain large numbers of functional gene-modified polyclonal T cells that can be used for adoptive immunotherapy.
Berger
Carolina
C
Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Blau
C Anthony
CA
Clackson
Tim
T
Riddell
Stanley R
SR
Heimfeld
Shelly
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D018106:CD28 Antigens / Q000378:metabolism*; D018929:Cell Culture Techniques; D019674:Genes, T-Cell Receptor beta; D006086:Graft vs Host Disease / Q000517:prevention & control; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008251:Macaca; D020800:Receptor, Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics; D016201:Receptors, Lymphocyte Homing / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D013937:Thymidine Kinase / Q000235:genetics / Q000494:pharmacology; D014161:Transduction, Genetic / Q000379:methods*
D018106:CD28 Antigens; D020800:Receptor, Nerve Growth Factor; D016201:Receptors, Lymphocyte Homing; D013937:Thymidine Kinase