title: In vitro comparison of three different chimeric receptor-modified effector T-cell populations for leukemia cell therapy.
abstract: The identification of the optimal T-cell effector subtype is a crucial issue for adoptive cell therapy with chimeric receptor-modified T cells. The ideal T cell population must be able to home toward tumor site, exert prolonged antitumoral activity, and display minimal toxicity against normal tissues. Therefore, we characterized the in vitro antitumoral properties of three effector T-cell populations: Epstein-Barr virus-specific cytotoxic T lymphocytes (EBV-CTLs), cytokine-induced killer (CIK) cells, and γ₉δ₂ T (GDT) cells, after transduction with a chimeric receptor specific for the CD33 antigen, broadly expressed on acute myeloid leukemia cells. EBV-CTLs, CIK, and GDT cells were generated and transduced with high efficiency with a retroviral vector coding for an anti-CD33-ζ chimeric receptor without alterations of their native phenotype. Anti-CD33-ζ chimeric receptor-redirected T cells displayed analogous in vitro chemotactic activity toward CXCL12. In addition, anti-CD33-ζ chimeric receptor-expressing EBV-CTLs, CIK, and GDT cells showed potent and similar cytotoxicity against several CD33⁺ leukemic targets both in short-term 4-hours-⁵¹chromium-release assays (mean killing vs primary leukemic cells at effector:target ratio of 5:1; 50%, 61%, and 50% for EBV-CTLs, CIK, and GDT cells, respectively) and in long-term assays, where they were cocultured with leukemic cells for 6 days on stromal mesenchymal cells (mean survival of primary leukemic cells at effector:target ratio of 1:100; 18%, 16%, and 29% for EBV-CTLs, CIK, and GDT cells, respectively). Moreover, all effector cells acquired consistent capability to proliferate in vitro after contact with CD33⁺ cells and to release high and comparable levels of immunostimulatory cytokines, while secreting similar low amount of immunoregulatory cytokines as the unmanipulated counterpart. Our results indicate that expression of an anti-CD33-ζ chimeric receptor potently and similarly increase the antileukemic functions of different effector T-cell subtypes, underlying the impossibility to identify a more potent T-cell population through in vitro analysis, and consistently with recent observations that have emerged from clinical trials with chimeric receptor-modified T cells, suggesting the need to perform such type of studies in the human setting.
authors.affiliation: Centro Ricerca M. Tettamanti, Clinica Pediatrica, Ospedale San Gerardo, Università Milano-Bicocca, Monza, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Pizzitola
authors.forename: Valentina
authors.initials: V
authors.lastname: Agostoni
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Cribioli
authors.forename: Martin
authors.initials: M
authors.lastname: Pule
authors.forename: Raphael
authors.initials: R
authors.lastname: Rousseau
authors.forename: Helene
authors.initials: H
authors.lastname: Finney
authors.forename: Alastair
authors.initials: A
authors.lastname: Lawson
authors.forename: Andrea
authors.initials: A
authors.lastname: Biondi
authors.forename: Ettore
authors.initials: E
authors.lastname: Biagi
authors.forename: Virna
authors.initials: V
authors.lastname: Marin
mesh_terms: D017252:CD3 Complex / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D064987:Cell- and Tissue-Based Therapy; D002634:Chemotaxis, Leukocyte / Q000276:immunology; D002648:Child; D046528:Chimerism; D005822:Genetic Vectors / Q000235:genetics; D018922:HL-60 Cells; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000276:immunology; D006801:Humans; D020014:K562 Cells; D007938:Leukemia / Q000276:immunology* / Q000628:therapy*; D008213:Lymphocyte Activation / Q000235:genetics / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014161:Transduction, Genetic
chemical_list: D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell