D'Elia, F.; Rizzi, M.; Manzi, R.; Perego, G.; Costagli, V.; Prada, A.; Santinami, M. Author Information

Background. Utilization of alpha-tumor necrosis factor (alpha-TNF) in clinical practice is limited by severe general side effects. Very promising results with low toxicity were reported with administration of alpha-TNF by isolation perfusion in extracorporeal circulation. Methods. From December 1991 to November 1992, 14 patients underwent perfusion with alpha-TNF (2-4 mg, total dose), gamma-interferon (1.5 X 10(6) IU], and melphalan (10 mg/l/perfused limb). Twelve patients presented intransit metastases of the limbs, one patient, a clear cell sarcoma of the hand, and one patient, a wide spindle cell carcinoma of the thigh. Perfusion lasted 90 minutes and was conducted in mild hyperthermia (38-40.5 degrees C, muscle temperature). Results. Nine complete regressions and four stable diseases were recorded. In one case, a reliable evaluation of response was not possible for diffused tissue necrosis. Five patients relapsed or progressed locally from 3 to 4 months after surgery, five presented distant localizations from 2 to 7 months after surgery, and one died of disease 6 months after perfusion. Twelve patients are alive, seven without evidence of disease. A septic-like shock syndrome was observed in all patients and required administration of dopamine, dobutamine, or noradrenaline. One patient died 30 days after perfusion from a multiorganfailure syndrome, likely due to (alpha-TNF. The follow-up time ranges from 4 to 15 months (median, 6). Conclusions. The preliminary, impressive results reported in other series were not completely confirmed in this study adopting the same treatment scheme. Further clinical experience and biologic data are needed to state the real efficacy of the approach and to reduce the severe general toxicity consistently associated with this type of treatment.

Human Gene TherapyVol. 5, No. 8 News and CommentsActive Immunization of Metastatic Melanoma Patients with Interleukin-4 Transduced, Allogeneic Melanoma Cells. A Phase I–II Study. University of Turin, ItalyPRINCIPAL INVESTIGATORS N. Cascinelli, R. Foà, G. Parmiani, CO-INVESTIGATORS F. Arienti, F. Belli, M. G. Bernengo, C. Clemente, M. P. Colombo, A. Guarini, M. T. Illeni, L. Mascheroni, C. Melani, A. Prada, and J. Sulé-SusoPRINCIPAL INVESTIGATORS N. CascinelliSearch for more papers by this author, R. FoàSearch for more papers by this author, G. ParmianiSearch for more papers by this author, CO-INVESTIGATORS F. ArientiSearch for more papers by this author, F. BelliSearch for more papers by this author, M. G. BernengoSearch for more papers by this author, C. ClementeSearch for more papers by this author, M. P. ColomboSearch for more papers by this author, A. GuariniSearch for more papers by this author, M. T. IlleniSearch for more papers by this author, L. MascheroniSearch for more papers by this author, C. MelaniSearch for more papers by this author, A. PradaSearch for more papers by this author, and J. Sulé-SusoSearch for more papers by this authorPublished Online:19 Mar 2008https://doi.org/10.1089/hum.1994.5.8-1059AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail FiguresReferencesRelatedDetailsCited byTargeting of IL-4 and IL-13 receptors for cancer therapyCytokine, Vol. 75, No. 1The evolving role of gene-based treatment in surgery4 November 2005 | British Journal of Surgery, Vol. 92, No. 12Molecular Strategies Interfering with Tumor Progression of Melanoma and Improving Anti-Tumor ImmunityVaccination for melanomaCurrent Oncology Reports, Vol. 2, No. 4HLA class I expression on human cancer cellsHuman Immunology, Vol. 61, No. 2Current Opinion in Oncology, Vol. 12, No. 2Vaccination of Melanoma Patients with Interleukin 4 Gene-Transduced Allogeneic Melanoma Cells Flavio Arienti, Filiberto Belli, Filomena Napolitano, Josep Sule-Suso, Arabella Mazzocchi, Gian Francesco Gallino, Alessandro Cattelan, Cristina Sanantonio, Licia Rivoltini, Cecilia Melani, Mario Paolo Colombo, Natale Cascinelli, Michele Maio, and Giorgio Parmiani6 July 2004 | Human Gene Therapy, Vol. 10, No. 18IL-4-Transduced Tumor Cell Vaccine Induces Immunoregulatory Type 2 CD8 T Lymphocytes That Cure Lung Metastases Upon Adoptive TransferThe Journal of Immunology, Vol. 163, No. 4Cancer VaccinesJournal of Clinical Oncology, Vol. 17, No. 3Strategies for Enhancing Tumor Immunogenicity (or how to transform a tumor cell in a Frankenstenian APC)Overview of melanoma vaccines: Active specific immunotherapy for melanoma patientsSeminars in Surgical Oncology, Vol. 14, No. 4GENE THERAPY FOR MELANOMA IN HUMANSHematology/Oncology Clinics of North America, Vol. 12, No. 3Clinical Approaches to Cancer Gene TherapyInterleukin-4Update on active specific immunotherapy with melanoma vaccinesJournal of Surgical Oncology, Vol. 66, No. 1Current Therapy Strategies for Malignant Melanoma with Special Regard to Immunotherapy with CytokinesCytokine Gene Transduction in the Immunotherapy of CancerImmunotherapy I: Cyclosine gene transfer strategiesCancer and Metastasis Reviews, Vol. 15, No. 3Gene therapy for cancer, the course aheadCancer and Metastasis Review, Vol. 15, No. 3A Human Melanoma Cell Line Transduced with an Interleukin-4 Gene by a Retroviral Vector Releases Biologically Active IL-4 and Maintains the Original Tumor Antigenic Phenotype Cecilia Melani, Josep Sulé-Suso, Flavio Arienti, Cristina Maccalli, Flavio Passerini, Mario P. Colombo, and Giorgio Parmiani19 March 2008 | Human Gene Therapy, Vol. 6, No. 11Tumor cells engineered to produce cytokines or cofactors as cellular vaccines: Do animal studies really support clinical trials?Cancer Immunology, Immunotherapy, Vol. 41, No. 5 Volume 5Issue 8Aug 1994 InformationCopyright 1994, Mary Ann Liebert, Inc.To cite this article:PRINCIPAL INVESTIGATORS N. Cascinelli, R. Foà, G. Parmiani, CO-INVESTIGATORS F. Arienti, F. Belli, M. G. Bernengo, C. Clemente, M. P. Colombo, A. Guarini, M. T. Illeni, L. Mascheroni, C. Melani, A. Prada, and J. Sulé-Suso.Active Immunization of Metastatic Melanoma Patients with Interleukin-4 Transduced, Allogeneic Melanoma Cells. A Phase I–II Study. University of Turin, Italy.Human Gene Therapy.Aug 1994.1059-1064.http://doi.org/10.1089/hum.1994.5.8-1059Published in Volume: 5 Issue 8: March 19, 2008PDF download

Prada, A.; Belli, F.; Costagli, V.; D'Elia, F.; Manzi, R.; Mascheroni, L.; Perego, G.; Rizzi, M.; Santinami, M.; Vaglini, M. Author Information

Belli, F.; Arienti, F.; Manzi, R.; Parmiani, G.; Persiani, L.; Rivoltini, L.; Santinami, M.; Santoro, N.; Prada, A.; Vaglini, M. Author Information

Prada, A.; Belli, F.; Costagli, V.; D'Elia, F.; Manzi, R.; Perego, G.; Rizzi, M.; Santinami, M.; Santoro, N.; Vaglini, M. Author Information

Prada, A.; Belli, F.; Costagli, V.; D'Elia, F.; Manzi, R.; Perego, G.; Persiani, L.; Rizzi, M.; Santinami, M.; Vaglini, M. Author Information

Freshly isolated tumor-infiltrating lymphocytes (TIL) from stage IV melanoma patients were cultured for 2 weeks with low doses of interleukin-2 (IL-2; 120 IU/ml), to select potentially for tumor-specific lymphocytes present in the neoplastic lesion, followed by high doses (6000 IU/ml) to achieve lymphocyte expansion. TIL were serially analyzed for their expansion, phenotype and cytotoxic activity against autologous and allogeneic tumor cells. A preferential lysis of autologous melanoma cells was obtained in long-term cultures of 7/13 cases (54%), while the remaining ones showed a major-histocompatibility-complex-unrestricted, lymphokine-activated-killer(LAK)-like activity at the time of in vivo injection. Sixteen patients with metastatic melanoma were infused with TIL (mean number: 6.8×109, range: 0.35 × 109−20 × 109) and IL-2 (mean dose: 130 × 106 IU, range: 28.8 × 106−231 × 106 IU); 1 complete and 3 partial responses were observed in 12 evaluable patients (response rate 33%). In all responding patients, injected TIL showed an in vitro preferential lysis of autologous tumor cells, while in no cases were TIL with LAK-like activity associated with a clinical response. The mean autologous tumor cytotoxic activity of TIL at the time of in vivo injection was significantly higher in responding patients in comparison to nonresponding ones, suggesting that a marked and preferential cytolysis of autologous tumor cells is associated with the therapeutic efficacy of TIL.

Arienti, P.; Belli, F.; Rivoltini, L.; Mascheroni, L.; Furlan, L.; Prada, A.; Parmiani, G.; Cascinelli, N. Author Information

Chemoresistant melanoma cells are known to be susceptible in vitro to lymphokine activated killer (LAK) cells. To obtain a high LAK/tumour cell ratio in vivo and avoid systemic toxicity due to interleukin-2 (IL-2), we used IL-2 plus LAK cells in the treatment of in transit melanoma metastases of the limbs by isolation perfusion (IP). In vivo immunological modifications induced by this immunotherapeutic approach were also analysed. Six patients previously treated with IP in extracorporeal circulation with tumour cytotoxic drugs and presently relapsing or not responding, were submitted to locoregional adoptive therapy consisting of 5 days systemic administration of IL-2 (Proleukin, EuroCetus) (9-12x10(6) IU/M2/day c.i.). Autologous LAK cells were derived from leukapheresis and subsequent in vitro stimulation with IL-2; LAK cells were then given along with IL-2 (120-2400 IU/ml of perfusion priming) to the affected limb by IP. In addition, 7-16x10(9) LAK cells were administered by systemic infusion the day after together with IL-2 (9-12x10(6) IU/m2/day) by c.i. for 5 days. All patients concluded the treatment without major toxicity. The analysis of circulating lymphocytes obtained from extracorporeal circuit at different times revealed rapid disappearance of LAK cells, suggesting their extravasation and/or endothelial adhesion in perfused tissues. Clinical responses included four partial and one complete response; another patient had stable disease. All patients are presently alive. Follow-up after IP ranges from 8 to 22 months.

Ten patients who underwent surgery (5 right hepatectomy and 5 colectomy) for cancer participated in a clinical controlled study. They were treated with buprenorphine (i.v. slow infusion) to relieve postsurgical pain. We found an increased urinary excretion of this drug in patients who underwent hepatectomy as compared with patients who underwent colectomy. However no differences in the occurrence of side-effects and/or in the therapeutic effect were observed between the two groups. We conclude that buprenorphine can be effectively and safely used also in patients with a resection of liver parenchyma.

From January 1987 to February 1988, 15 stage IV melanoma patients were treated with two courses of bolus injection of rIL-2 plus LAK cell infusions at the National Cancer Institute of Milan. The original treatment regimen included a first course of rIL-2 administration (400 micrograms/m2 bolus injection 3 times a day [TID] for 4 days) and a second course of rIL-2 administration (800 micrograms/m2 bolus injection TID for 7 days) separated by 4 consecutive daily leukaphereses. Autologous lymphokine activated killer (LAK) cells were reinfused into each patient on three occasions during the second period of rIL-2 administration. Due to the appearance of grade III-IV neurological, hepatic and cardiopulmonary toxicity, 7 patients discontinued dosing before the end of treatment, one patient desired to be withdrawn and one patient died from rapidly progressive disease, although complications of rIL-2 administration may have contributed to her death. Only 6 patients completed the schedule without evidence of major intolerance, even though the planned dose during the second course of rIL-2 was reduced to 400 micrograms/m2. The complete duration of treatment ranged from 11 to 19 days. The total dose of rIL-2 injected ranged from 12.6 to 30.4 mg. The number of infused LAK cells ranged from 15.5 x 10(9) to 60 x 10(9)/patient. Two of the 14 evaluable patients showed a minor anti-tumor response. In 5 patients new metastases in other sites were documented from 2 to 5 months after completion of dosing. No apparent association was found between progression of the disease (or the appearance of new metastases) and the total dose of rIL-2 injected, the number of LAK cells administered or the number of days of treatment. By December 1988, all patients had died of their disease in a period ranging from 3 to 14 months from the last injection of rIL-2. The lack of significant clinical responses in this study and the high toxicity of this treatment lead us to conclude that at least as far as melanoma patients are concerned, adoptive immunotherapy with rIL-2 plus LAK cells (as described here) is not a justifiable treatment option unless new evidence presents itself.

Toxicity and clinical effects of a new brand of recombinant interleukin 2 (rIL2, Bioleukin™, Biogen, Geneva) were evaluated by a phase I study in 12 patients with stage III melanoma. Escalating doses from 100 μg/m2 to 800 μg/m2 were administered thrice a day with bolus injections given via a peripheral venous catheter for up to a maximum of 7 days. All patients showed malaise, fever and chills and mild gastrointestinal side effects. A modest electrolyte imbalance (hypocalcemia and hypokalemia) was detected in all 12 patients. Renal toxicity as judged by serum creatinine was not observed, and hepatic toxicity was moderate with the possible exception of one patient who had an unclear previous history of liver dysfunction. Mild, transient leukopenia was found in several patients, whereas thrombocytopenia developed in 4 patients; no anemia was observed. Cutaneous rash was seen in half of the patients treated. Fluid retention was minimal, with a weight gain associated to treatment that never exceeded 10% of pretreatment body weight. Electrocardiographic alterations developed in 2 patients in the form of ventricular and supraventricular extrasystoles. In 2 patients given the highest dose of rIL2, a significant reduction in transfer lung factor for carbon monoxide was seen, indicating alterations in pulmonary functions. Other dose-related toxicities were thrombocytopenia and malaise. All side effects disappeared 2-3 days after the end of rIL2 administration. No major responses were seen in association with the 4-8 days of treatment given in this study.

The aim of this paper is to evaluate the effectiveness of DTIC when employed at a local level in hyperthermic antiblastic perfusion (HAP) for stage IIIA-IIIAB melanoma patients. Twenty-seven consecutive patients have been treated at the National Cancer Institute of Milan from October 1983 to June 1985. All the patients were submitted to HAP at 40 degrees for 60' with DTIC at the dosage of 2.5 g/m2 [corrected] for lower extremities and 1.5 g/m2 [corrected] for upper extremities. We observed a complete local response in three patients and a partial local response 50% in seven patients, 10 patients has a response less than 50% and 4 patients did not show any response. After surgical removal of the residual tumor when possible, 14 patients are alive without detectable disease while 11 are alive with disease and two dead for progression. No serious complications were observed. These data indicate that DTIC seems able to obtain in HAP, results superimposable to L-PAM without any significant toxicity.

Regional perfusion in extracorporeal circulation is finding more and more applications in the treatment of extremities’ tumors. In order to avoid any influence from surgical variables rises the necessity of a standardization of the technique. In this paper we propose a new surgical technique with subsequent results based on the experience at the National Cancer Institute of Milan on 89 consecutive patients treated by regional hyperthermic antiblastic perfusion without significant complications.

From February 1982 to December 1983, 42 patients affected by neoplasms of the limbs were treated at the Istituto Nazionale Tumori of Milan by hyperthermic antiblastic perfusion in extracorporeal circulation at the temperature of 40-41 degrees C for 1 h. Thirty-two were affected by melanoma, 4 by osteogenic sarcoma, 2 by squamous-cell carcinoma, 1 by liposarcoma, 1 by hemangiopericytoma, 1 by clear-cell sarcoma and 1 by Kaposis's sarcoma. As regards the immediate response, a complete plus partial remission rate of 88% without any major complication was obtained. The follow-up period is too short for any considerations about overall survival. However, because of these good clinical results we consider this method able to locally control the evolution of neoplasms of the extremities, allowing in many cases a limb salvage.

Two patients with extensive squamous cell carcinoma of the lower extremities, candidate for demolitive surgery, were treated by hyperthermic antiblastic perfusion in extracorporeal circulation. The temperature reached was 41 °C and the drug used was methotrexate at the dosage of 500 mg. Radical excision of the ulcer was possible in the 2 patients. Both tumors underwent extensive necrosis, and histology done 1 month after perfusion on surgical specimens showed limited areas of residual malignancy. These 2 patients suggest that hyperthermic antiblastic perfusion may be a limb salvage procedure in the multimodal management of extended squamous cell carcinoma of the extremities.