Vaccination with Autologous Tumor-derived Heat-Shock Protein Gp96 after Liver Resection for Metastatic Colorectal Cancer 1 Vincenzo Mazzaferro, 2 Jorgelina Coppa, Matteo G. Carrabba, Licia Rivoltini, Marcello Schiavo, Enrico Regalia, Luigi Mariani, Tiziana Camerini, Alfonso Marchiano `, Salvatore Andreola, Roberto Camerini, Marco Corsi, Jonathan J. Lewis, Pramod K. Srivastava, and Giorgio Parmiani Units of Hepatobiliary and Gastro-Pancreatic Surgery [V. M., J. C., M. S., E. R.], Immunotherapy of Human Tumors [M. G. C., L. R., G. P.], Biomedical Statistics [L. M., T. C.], Radiology [A. M.], and Pathology [S. A.], Istituto Nazionale per lo Studio e la Cura Tumori, 20133 Milan, Italy; Sigma-Tau S.p.A., 00040 Rome, Italy [R. C., M. C.]; Antigenics Inc., New York, New York 10111 [J. J. L.]; and Department of Microbiology and Immunology, University of Connecticut, Farmington, Connecticut 06030 [P. K. S.] ABSTRACT Purpose: Heat shock proteins (HSP) from tumor cells contain the gp96 polypeptide associated with cancer-specific antigenic peptides. Mice that are immunized with HSP/ peptide-complex (HSPPC) derived from cancer tissue reject tumor from which HSPs are purified. We tested in humans whether vaccination with HSPPC-gp96 (Oncophage) from autologous liver metastases of colorectal carcinoma induces cancer-specific T-cell responses in patients rendered disease free by surgery. Experimental Design: Twenty-nine consecutive patients underwent radical resection of liver metastases [Memorial Sloan-Kettering Cancer Center (MSKCC) score 1–3 (good prognosis), 18 patients; score 4 –5 (bad prognosis), 11 pa- tients] and received autologous tumor-derived HSPPC-96. Two vaccine cycles were administered (four weekly injec- tions followed by four biweekly injections after 8 weeks). Class-I HLA-restricted, anti-colon cancer lines T-cell re- sponse was measured by ELISPOT assay on peripheral blood mononuclear cells (PBMCs) obtained before and after vaccination. Feasibility, safety, and possible clinical benefits were also evaluated. Results: Either a de novo induced or a significant in- crease of preexisting class I HLA-restricted T-cell-mediated anti-colon cancer response was observed in 15 (52%) of 29 patients. Frequency of CD3, CD45RA, and CCR7- T lymphocytes increased in immune responders. No relevant toxicity was observed. As expected, patients with good prog- nosis had a significantly better clinical outcome than those with poor prognosis [2-year overall survival (OS), 89 versus 64%, P  0.001; disease-free survival (DFS), 46 versus 18%, P  0.001]. Patients with immune response had a statisti- cally significant clinical advantage over nonresponding sub- jects (2-year OS, 100% versus 50%, P  0.001; DFS, 51% versus 8%, P  0.0001). Occurrence of immune response led to better tumor-free survival, whatever the predicted prog- nosis was (hazard ratio, 0.11– 0.12 with/without stratifica- tion; P  0.0012– 0.0003). Conclusions: HSPPC-96 vaccination after resection of colorectal liver metastases is safe and elicits a significant increase in CD8 T-cell response against colon cancer. In this limited number of patients, two-year OS and DFS were significantly improved in subjects with postvaccination an- titumor immune response, independently from other clinical prognostic factors. INTRODUCTION CRC, 3 one the most common malignancies in humans, has frequent metastatic spread to the liver. Indeed, liver metastases are detectable in 15–25% of patients at the time of first diag- nosis, progressing to about 60% of patients during their disease course (1, 2). Occurrence of liver metastases parallels the stage of primary CRC and represents the leading cause of death in advanced stages, with autopsy series showing liver metastases in more than 80% of patients who died from CRC (3). Surgical resection remains the primary treatment, although complete tumor removal from the liver is achieved in only 20 – 40% of the patients (4) resulting in a 5-year OS of 40% (1, 5) and tumor recurrence of 59 – 81% according to different stages at the time of surgery (4, 6). A large retrospective analysis has identified several independent determinants of Received 1/2/03; revised 2/21/03; accepted 2/24/03. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Supported by a grant from Sigma-Tau i.f.r., S.p.A., Rome. V. M. is recipient of the grant “New therapeutic strategies in liver tumors” by the Italian Association for Cancer Research (AIRC). P. K. S. is supported by a NIH grant and through a sponsored research agreement with Antigenics Inc., in which he has a substantial financial interest. R. C. and M. C. are employees of the Sigma-Tau i.f.r. S.p.A., and J. J. L. is an employee of Antigenics Inc. Partially presented at the 37th Annual Meeting of the American Society of Clinical Oncology (ASCO), San Francisco, CA, May 14, 2001. 2 To whom requests for reprints should be addressed, at Department of Surgery, Hepatobiliary and Gastro-Pancreatic Unit, Istituto Nazionale Tumori, Via Venezian 1, 20133 Milano, Italy. Phone: 39-02-2390-2760; Fax: 39-02-2390-.3259; E-mail: vincenzo.mazzaferro@istitutotumori.mi.it. 3 The abbreviations used are: CRC, colorectal carcinoma; APC, antigen- presenting cell; CCR7, chemokine receptor 7; CEA, carcinoembryonic antigen; EP-CAM, epithelial cell adhesion molecule; DC, dendritic cell; DFS, disease-free survival; HSP, heat shock protein; HSPPC, HSP/ peptide-complex; mAb, monoclonal antibody; NK, natural killer; OS, overall survival; PBMC, peripheral blood mononuclear cell; MSKCC, Memorial Sloan-Kettering Cancer Center; ELISPOT, enzyme-linked immunospot; ATCC, American Type Culture Collection. 3235 Vol. 9, 3235–3245, August 15, 2003 Clinical Cancer Research Cancer Research. on November 26, 2021. © 2003 American Association for clincancerres.aacrjournals.org Downloaded from