Exome Sequencing to Predict Neoantigens in Melanoma
Abstract The ability to use circulating peripheral blood cells and matched tumor sequencing data as a basis for neoantigen prediction has exciting possibilities for application in the personalized treatment of cancer patients. We have used a high-throughput screening approach, combining whole-exome sequence data, mRNA microarrays, and publicly available epitope prediction algorithm output to identify mutated proteins processed and displayed by patient tumors and recognized by circulating immune cells. Matched autologous melanoma cell lines and peripheral blood mononuclear cells were used to create mixed lymphocyte tumor cell cultures, resulting in an expansion of tumor-reactive T cells to u…
Temozolomide- and fotemustine-induced apoptosis in human malignant melanoma cells: response related to MGMT, MMR, DSBs, and p53
Malignant melanomas are highly resistant to chemotherapy. First-line chemotherapeutics used in melanoma therapy are the methylating agents dacarbazine (DTIC) and temozolomide (TMZ) and the chloroethylating agents BCNU and fotemustine. Here, we determined the mode of cell death in 11 melanoma cell lines upon exposure to TMZ and fotemustine. We show for the first time that TMZ induces apoptosis in melanoma cells, using therapeutic doses. For both TMZ and fotemustine apoptosis is the dominant mode of cell death. The contribution of necrosis to total cell death varied between 10 and 40%. The O(6)-methylguanine-DNA methyltransferase (MGMT) activity in the cell lines was between 0 and 1100 fmol m…
Generation of CD8+T cells expressing two additional T-cell receptors (TETARs) for personalised melanoma therapy
Adoptive T-cell therapy of cancer often fails due to the tumor cells' immune escape mechanisms, like antigen loss or down-regulation. To anticipate immune escape by loss of a single antigen, it would be advantageous to equip T cells with multiple specificities. To study the possible interference of 2 T-cell receptors (TCRs) in one cell, and to examine how to counteract competing effects, we generated TETARs, CD8(+) T cells expressing two additional T-cell receptors by simultaneous transient transfection with 2 TCRs using RNA electroporation. The TETARs were equipped with one TCR specific for the common melanoma antigen gp100 and one TCR recognizing a patient-specific, individual mutation of…
Pseudopathological vertebral changes in a young individual from Herculaneum (79 C.E.)
Post mortem abnormal modification of bone are known as pseudopathologies. The geochemical characteristic of the burial soil and/or the presence of biological agents may produce marked changes in bone preservation. This could be the case for a young individual, E74, from Herculaneum, which was a Roman town near Naples completely destroyed by the volcanic eruption of Mt. Vesuvius in 79 CE. E74 is an incomplete skeleton of a male individual of 7-8 years of age. Its second and third cervical vertebrae, the eighth thoracic vertebra and the first lumbar vertebra show a septum dividing the vertebral foramen. This condition could be diagnosed as diastematomyelia that consists of the splitting of th…