Cancer Immunol Immunother (1987) 24:76-85 ancer mmunolggy mmunotherapy © Springer-Verlag 1987 Tumor-derived interleukin-2-dependent lymphocytes in adoptive immunotherapy of lung cancer Richard L. Kradin, Lenora A. Boyle, Frederic I. Preffer, Ronald J. Callahan, Martha Barlai-Kovach, H. William Strauss, Steven Dubinett and James T. Kurnick Medicine, Radiology and Pathology, Departments of the Massachusetts General Hospital and the Harvard Medical School, Boston, Massachusetts, USA Summary. A trial of adoptive immunotherapy was per- formed in which long-term cultured, interleukin-2 (IL2)-dependent T-lymphocytes were administered to pa- tients with metastatic adenocarcinoma of the lung. Lym- phocytes were isolated from explants of cancer tissues that were cultured in medium with recombinant IL-2. These T- cells expressed surface markers of activation, and killed a broad panel of tumor targets. Intravenously injected 11 qn- dium-labeled T-cell blasts distributed primarily to lungs, liver, and spleen. Despite a paucity of infused lymphocytes detected by external imaging at sites of tumor, five of se- ven patients showed reduction of their cancers. However, in no case was greater than 50% reduction of total tumor burden achieved. Evidence of increased delayed cutaneous hypersensitivity to protein antigens was observed in three patients following therapy. We conclude that long-term cultured tumor-derived T-cells can be transferred safely into humans and that these cells may be capable of en- hancing immune responses and mediating tumor reduc- tion in vivo. Introduction Tumor cells express surface antigens that serve as targets for cell-mediated immunological responses [22]. It has been repeatedly demonstrated that animals that have been immunized against a tumor can specifically reject subse- quent challenges by that tumor [23]. Rejection in vivo is mediated by mononuclear cells and specificity for tumor is conferred by T-lymphocytes [3, 12]. Lymphocytes sensi- tized to syngeneic tumor in vitro can effectively produce regression and cure of certain advanced murine tumors [4, 18, 31, 38, 40]. In addition to antigen-specific T-lympho- cytes, natural killer (NK) cells [21] and other nonspecific cytotoxic cells [27, 34] have been suggested to play a role in reducing tumor in vivo. Recently, Rosenberg et al. have reported clinical responses in some human cancers by the transfer of interleukin-2 (IL-2)-stimulated peripheral blood lymphocytes that display broad antitumor specifici- ties [39]. T-lymphocytes that are isolated from human tumors are enriched for specific reactivities to their autologous tu- mor when examined in vitro [24, 44, 45]. The adoptive Offprint requests to: Richard Kradin, Cox-5 Immunopathology Unit Massachusetts General Hospital Boston, MA 02114, USA transfer of sensitized, tumor-specific T-lymphocytes, how- ever, has not been applied to the treatment of human can- cers due to difficulties in the isolation and propagation of large numbers of immune T-cells. T-lymphocytes can be isolated from autologous tumor tissues when fresh tumor explants are cultured in medium with IL-2 [24]. T-cells re- trieved in this manner secrete lymphokines, bind to tumor cells, kill autologous and allogeneic cancer cells and the K562 erythroleukemia cell line. These T-lymphocytes also display enhanced killing of their autologous tumors be- yond what is observed for peripheral blood lymphocytes from the homologous donors. We report the results of an immunotherapeutic trial in metastatic adenocarcinoma of lung using the adoptive transfer of autologous, long-term cultured, tumor-derived T-cells. Five of seven patients who received adoptive transfers of autologous T-lymphocytes without adjuvant IL-2, showed a reduction in the size of their tumors. The antitumor effects of immunocompetent T-cells isolated from tumor tissues suggest that these cells may be of thera- peutic value in human lung cancer. Experimental methods Patient population. Seven patients with metastatic adeno- carcinoma of lung were studied. The criteria for entry into the trial included the presence of biopsy-proven metastatic adenocarcinoma of the lung that was not amenable to sur- gical extirpation. Patients were excluded from the trial if they were(l) >70 years old, (2) had arterial pO2 <60 mm Hg, (3) a creatinine >3.0mg/100ml, (4) bilirubin >3.0 rag/100 ml, (5) active arthritis or autoimmune dis- ease, or (6) were receiving steroids. All patients signed an approved informed consent form prior to entrance into the trial. Four of the seven patients had completed courses of radiation to their primary lung tumor and mediastinum at least 1 month prior to treatment. One patient (D) complet- ed a single course of chemotherapy, without achieving a clinical response, prior to beginning immunotherapy. Three (B, C, G) received palliative radiotherapy to painful boney lesions during their course of immunotherapy (Table 1). Irradiated sites in bone were excluded from posttreatment evaluation. Lymphocyte harvest and culture. Recombinant IL-2 was a generous gift of the Genetics Institute (Boston, Mass.). In