FACS analysis was performed with a FACSCalibur Flow cytometer (Becton Dickinson, Heidelberg, Germany) using CellQuest Pro and WinMDI software. Unstimulated PBMC and
PBMC after incubation with allogeneic EpCAM+ HT-29 (ATCC Nr. CCL-244) or HER2/neu+ SK-BR-3 carcinoma cells (ATCC Nr. HTB-30) were used as negative controls. Clinical patient evaluation/toxicity and safety evaluation Careful patient monitoring was applied throughout the study. Clinical evaluation, including medical history and general physical exam, was performed at baseline and defined days during treatment (day of trAb infusion and the following day; day of restimulation P505-15 supplier and the following day). Patients were monitored for adverse events according to the National
Cancer Institute common toxicity criteria during each visit. Standard laboratory parameters and vital signs were tested before and after treatment Laboratory testing included complete blood count, electrolytes, creatinine, bilirubin, transaminases, and tumor marker (CA19-9 for gastric carcinoma, CA125 for ovarian carcinoma, CEA and CA125 for CUP). In addition, patients during trAb therapy were daily monitored for systemic cytokine responses. Blood samples were taken before, 24 hours and 48 h after every trAb application. Serum levels of IL-6, TNF-α, and sIL-2R were measured by ELISA (Biosource, Fleurs, Belgium). Immune reaction to mouse https://www.selleckchem.com/products/JNJ-26481585.html IgG was assessed by ELISA measurement of human anti-mouse antibody reaction (HAMA) before and 4 weeks after therapy. Response was evaluated by computed tomography two to three months
after trAb treatment and every two to three months until tumor progression. Statistical analysis Analysis of cytokine levels was performed using the Wilcoxon signed rank test. Correlation analysis was done by the chi-square contingency analysis. All tests were calculated by SAS statistical software using a Windows XP computer system. Results Patients’ characteristics Nine patients were treated between February click here 2005 and December 2007. Prior to study treatment, 6 patients underwent surgical resection with curative intent, 8 patients received chemotherapy. Four patients presented with synchronous PC, whereas five developed PC after surgery and chemotherapy. One patient was diagnosed with PC of carcinoma of unknown primary (CUP) during elective laparoscopic cholecystectomy. The patients’ demographic and primary treatment parameters are listed in Table 1. Table 1 Patients’ characteristics Pat. Age Sex Tumor entity TNM stage primary Surgical therapy primary tumor Chemotherapy before trAb Radiation before trAb EpCAM expression HER2/neu expression A 31 f Gastric pT4pN3M0 Gastrectomy + + + + B 64 f Ovarian pT3pN0M0 Adnexectomy, resect. of liver met.