p. administration and restimulation with trAb in patients with PC. Patients and methods Objectives and study approval This study was designed as a sequential dose-escalating, feasibility study for compassionate use of trAb in the induction of tumor immunity. The study was carried out according to the principles of the Declaration of Helsinki and good clinical practice guidelines. It was approved by the Ethics committee of the selleck Ludwig-Maximilians-University, Munich, Germany. Informed consent was obtained from all patients prior to treatment. Patients Patients enrolled in this study had histologically confirmed diagnosis of PC. Inclusion

criteria were Karnofsky performance status ≥ 60%, white blood cell count > 2000/mm3 and a relative T-cell count > 10%. Exclusion criteria included prior immunotherapy, significant heart disease or arrhythmia,

known allergic reactions or XL184 supplier autoimmune disease, significant liver, kidney, pulmonary or haematological disease, acute or chronical infection and paracentesis of malignant ascites > 1000 ml within 30 days before treatment. Patients were included independent of any prior conventional therapy, i.e. chemotherapy, radiation or tumor surgery. An interval of more than 30 days between any chemotherapy and the start of the trAb therapy was required. A recovery interval of at least 7 days after abdominal surgery with laparotomy was mandatory. All patients had a surgical procedure (explorative laparotomy or laparoscopy, resection of intra-abdominal metastases), where isolation of autologous tumor samples was possible. Isolation and storage of autologous tumor cells Autologous tumor samples were taken during surgery (explorative laparotomy or laparoscopy, resection of intra-abdominal metastasis). The surgical procedure was independent from study inclusion. Patients were only included if more than 5 × 106 autologous tumor cells were successfully

isolated, and if EpCAM antigen or HER2/neu antigen was found on > 10% of all viable cells Sulfite dehydrogenase from autologous tumor cell preparations. Analysis of autologous tumor cells was performed by immunohistochemical APAAP staining [23] using the antibodies HO3 (anti-EpCAM; mouse IgG2a, TRION Pharma) or C215 (anti EpCAM; mouse IgG2a; kindly provided by M. Dohlsten, Pharmacia, Uppsala, Sweden) for EpCAM or 2502A (anti Her2/neu; mouse IgG2a; Trion Pharma, Munich, Germany) for HER2/neu. After surgical resection autologous tumor probes were dissected into 2–3 mm3 pieces which were then immersed in RPMI 1640 medium (containing 0.05% Collagenase type 4, 0.02% DNAse type 1, Penstrep, Gentamycin and Amphotericin B; all reagents from Invitrogen, Carlsbad, California). This mixture was incubated overnight at 37°C and filtered through a flexible grid to exclude undigested tissue fragments.