title: Manufacture of gene-modified human T-cells with a memory stem/central memory phenotype.
abstract: Advances in genetic engineering have made it possible to generate human T-cell products that carry desired functionalities, such as the ability to recognize cancer cells. The currently used strategies for the generation of gene-modified T-cell products lead to highly differentiated cells within the infusion product, and on the basis of data obtained in preclinical models, this is likely to impact the efficacy of these products. We set out to develop a good manufacturing practice (GMP) protocol that yields T-cell receptor (TCR) gene-modified T-cells with more favorable properties for clinical application. Here, we show the robust clinical-scale production of human peripheral blood T-cells with an early memory phenotype that express a MART-1-specific TCR. By combining selection and stimulation using anti-CD3/CD28 beads for retroviral transduction, followed by expansion in the presence of IL-7 and IL-15, production of a well-defined clinical-scale TCR gene-modified T-cell product could be achieved. A major fraction of the T-cells generated in this fashion were shown to coexpress CD62L and CD45RA, and express CD27 and CD28, indicating a central memory or memory stemlike phenotype. Furthermore, these cells produced IFNγ, TNFα, and IL-2 and displayed cytolytic activity against target cells expressing the relevant antigen. The T-cell products manufactured by this robust and validated GMP production process are now undergoing testing in a phase I/IIa clinical trial in HLA-A*02:01 MART-1-positive advanced stage melanoma patients. To our knowledge, this is the first clinical trial protocol in which the combination of IL-7 and IL-15 has been applied for the generation of gene-modified T-cell products.
authors.affiliation: 1 Division of Immunology, The Netherlands Cancer Institute , 1066 CX Amsterdam, The Netherlands .
authors.forename: Raquel
authors.initials: R
authors.lastname: Gomez-Eerland
authors.forename: Bastiaan
authors.initials: B
authors.lastname: Nuijen
authors.forename: Bianca
authors.initials: B
authors.lastname: Heemskerk
authors.forename: Nienke
authors.initials: N
authors.lastname: van Rooij
authors.forename: Joost H
authors.initials: JH
authors.lastname: van den Berg
authors.forename: Jos H
authors.initials: JH
authors.lastname: Beijnen
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Uckert
authors.forename: Pia
authors.initials: P
authors.lastname: Kvistborg
authors.forename: Ton N
authors.initials: TN
authors.lastname: Schumacher
authors.forename: John B A G
authors.initials: JB
authors.lastname: Haanen
authors.forename: Annelies
authors.initials: A
authors.lastname: Jorritsma
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D060846:Cell Engineering / Q000379:methods; D049109:Cell Proliferation; D002986:Clinical Trials as Topic; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007156:Immunologic Memory / Q000235:genetics*; D007371:Interferon-gamma / Q000235:genetics / Q000276:immunology; D019409:Interleukin-15 / Q000494:pharmacology; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology; D015851:Interleukin-7 / Q000494:pharmacology; D058965:MART-1 Antigen / Q000235:genetics / Q000276:immunology; D008545:Melanoma / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology*; D012190:Retroviridae / Q000235:genetics; D012878:Skin Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000637:transplantation; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology
chemical_list: D015703:Antigens, CD; C569274:IL15 protein, human; C508605:IL7 protein, human; D019409:Interleukin-15; D007376:Interleukin-2; D015851:Interleukin-7; D058965:MART-1 Antigen; D011948:Receptors, Antigen, T-Cell; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma