Ex vivo expansion protocol for human tumor specific T cells for adoptive T cell therapy.
Adoptive T cell therapy is a promising treatment strategy for patients with different types of cancer. The methods used for generation of high numbers of tumor specific T cells usually require long-term ex vivo culture, which frequently lead to generation of terminally differentiated effector cells, demonstrating low persistence in vivo. Therefore, optimization of protocols for generation of T cells for adoptive cell therapy is warranted. The aim of this work was to develop a protocol for expansion of antigen-specific T cells using Dynabeads CD3/CD28 to obtain T cells expressing markers important for in vivo persistence and survival. To achieve high numbers of antigen-specific T cells following expansion, we have tested the effect of depleting regulatory T cells using Dynabeads CD25 and including a pre-stimulation step with peptide prior to the non-specific expansion with Dynabeads. Our data demonstrate that virus- and tumor specific T cells can be expanded to high numbers using Dynabeads CD3/CD28 following optimization of the culture conditions. The expansion protocol presented here results in enrichment of antigen-specific CD8(+) T cells with an early/intermediate memory phenotype. This is observed even when the antigen-specific CD8(+) T cells demonstrated a terminal effector phenotype prior to expansion. This protocol thus results in expanded T cells with a phenotypic profile which may increase the chance of retaining long-term persistence following adoptive transfer. Based on these data we have developed a cGMP protocol for expansion of tumor specific T cells for adoptive T cell therapy.
Rasmussen
Anne-Marie
AM
Department of Immunology, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway. annemra@rr-research.no
Borelli
Gabriel
G
Hoel
Hanna Julie
HJ
Lislerud
Kari
K
Gaudernack
Gustav
G
Kvalheim
Gunnar
G
Aarvak
Tanja
T
D019264:Adoptive Transfer; D015703:Antigens, CD / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002470:Cell Survival; D005260:Female; D006801:Humans; D007156:Immunologic Memory; D018189:Immunomagnetic Separation; D008297:Male; D009369:Neoplasms / Q000276:immunology / Q000628:therapy
D015703:Antigens, CD