∗ Contributed equally as senior authors. Correspondence: Daniela Montagna, Laboratorio di Immunologia e Trapianti, Dipartimento di Scienze Pediatriche, Università degli Studi di Pavia, Fondazione IRCCS Policlinico San Matteo, P.le Golgi 2,27100 Pavia, Italy. E-mail: d.montagna@smatteo.pv.it (Received 14 January 2011; accepted 28 July 2011) Feasibility and safety of adoptive immunotherapy with ex vivo-generated autologous, cytotoxic T lymphocytes in patients with solid tumor DANIELA MONTAGNA 1,2 , ILARIA TURIN 2 , ROBERTA SCHIAVO 3 , ENRICA MONTINI 1 , NADIA ZAFFARONI 4 , RAFFAELLA VILLA 4 , SIMONA SECONDINO 3 , ILARIA SCHIAVETTO 3 , LAURA CALIOGNA 2 , FRANCO LOCATELLI 1,5∗ , VIRGINIA LIBRI 6 , ANDREA PESSION 6 , ROBERTO TONELLI 7 , RITA MACCARIO 2,8∗ , SALVATORE SIENA 3∗ & PAOLO PEDRAZZOLI 3,9,10 1 Dipartimento di Scienze Pediatriche, Università di Pavia, Pavia, Italy, 2 Laboratorio di Immunologia dei Trapianti, Oncoematologia Pediatrica, Fondazione IRCCS Policlinico S. Matteo, Pavia, Italy, 3 SC Oncologia Medica Falck, Ospedale Niguarda Ca’ Granda, Milano, Italy, 4 Dipartimento di Oncologia Sperimentale, Fondazione IRCCS Istituto Nazionale dei Tumori, Milano, Italy, 5 Dipartimento di Oncologia-Ematologia Pediatrica, IRCCS Ospedale Bambino Gesù, Roma, Italy, 6 Dipartimento di Ematologia e Oncologia Pediatrica, Università di Bologna, Bologna, Italy, 7 Dipartimento di Farmacologia, Università di Bologna, Bologna, Italy, 8 Cell Factory, Lab. Ricerca, Fondazione IRCCS Policlinico S. Matteo, Pavia , Italy, 9 Gruppo Italiano di Trapianto di midollo, cellule staminali ematopoietiche e terapia cellulare (GITMO), and 10 SC Oncologia, Fondazione IRCCS Policlinico S. Matteo, Pavia, Italy Abstract Background aims. Adoptive T-cell therapy with tumor-specific T cells has emerged as a potentially useful approach for treating patients with advanced malignancies. We have demonstrated previously the feasibility of obtaining large numbers of autologous anti-tumor-specific cytotoxic T lymphocytes (CTL) generated by stimulation of patients’ peripheral blood mononuclear cells with dendritic cells pulsed with apoptotic tumor cells. Methods. Six patients with progressing metastatic solid tumors (one renal cell carcinoma, two ovarian cancers, two extraosseous peripheral neuroectodermal tumors, one soft tissue sarcoma) not eligible for conventional therapies were treated with adoptive immunotherapy. Anti-tumor CTL, proven to be reactive in vitro against patient tumor cells, but not against normal cells, were infused following lymphodepleting chemotherapy administered to favor T-cell proliferation in vivo. Results. Patients received a median of nine CTL infusions (range 2–19). The median number of CTL administered per infusion was 11  10 8 (range 1–55  10 8 ). No patient expe- rienced acute or late adverse events related to CTL infusion, even when large numbers of cells were given. Post-infusion laboratory investigations demonstrated an increase in the frequency of circulating anti-tumor T-cells and, in patients with a longer follow-up receiving two CTL infusions/year, a stabilization of these values. Conclusions. Our study demonstrates that autologous ex vivo-generated anti-tumor CTL can be administered safely in patients with advanced solid tumors and can improve the immunologic reactivity of recipients against tumor. These preliminary results provide a rationale for evaluating the clinical efficacy of this immunotherapeutic approach in phase I/II studies. Key Words: adoptive T-cell therapy, autologous tumor cells, cytotoxic T lymphocytes, lymphodepletion, solid tumors Introduction Adoptive T-cell immunotherapy is thought to be a means for enhancing anti-tumor immunity with the aim of controlling tumor growth and/or preventing tumor recurrence.Therapies based on adoptive trans- fer of T cells provide an opportunity to overcome some of the mechanisms of tumor escape, by infusing ex vivo-selected, tumor-reactive T lymphocytes and by manipulating the host environment into which T cells are introduced. Successful therapy depends on the type of T cells transferred and their ability to reach the tumor site, to display effector functions and to persist over time (1). The anti-tumor T lympho- cytes transferred can be divided into at least two Cytotherapy, 2012; 14: 80–90 ISSN 1465-3249 print/ISSN 1477-2566 online © 2012 Informa Healthcare DOI: 10.3109/14653249.2011.610303