y undergo a process of maturation and e press large amounts of human leukocyte Ags peptide com ple es at their surface. To achieve their function, DCs must arrive into the lymph nodes in response to several chemoattracting signals that bind specific cell surface receptors e pressed by DCs during the maturation proc ess, such as CCR7. selleck chemical Once DCs have arrived into secondary lymphoid organs, they can stimulate na ve T cells. A common strategy used for vaccine preparation is to load DCs with e ogenous peptides from tumor associated Ags on empty HLA class I molecules. This approach, however, has the limitations of peptide restric tion to a given haplotype and the induction of responses to only one or few defined Ags. In order to use a broader spectrum of known and yet unknown Ags for DCs load ing, the approach of whole tumor cells is preferred.

We and others have demonstrated that when murine DCs that had phagocytosed apoptotic B16 melanoma cells were used as vaccines, they were able to induce an effective, long term protection against challenge with live B16 cells. Since the induction of CD8 cytoto ic T lymphocytes appears to play a central role in the process of pro tective immunity, only cross presentation of tumor Ags acquired from whole tumor cells would confer effective antitumor immunity. Several authors have demonstrated in murine models and in humans. that when DCs engulf apoptotic cells, Ags can be cross presented for the generation of HLA class I peptide comple es, allow ing the induction of specific CTLs.

However, some con flicting findings have been reported in the human, such as the lack of DCs maturation upon phagocytosis of apop totic cells, so the fate and immunogenic potential of DCs that have internalized melanoma apoptotic cells or their debris remain an open issue. Several studies have used tumor cells virally GSK-3 transduced with TAAs or tumor cells apoptotized after infection with recombinant viruses encoding melanoma associated Ags but few of these have evaluated the specific cross presentation of native melanoma Ags present in apoptotic tumor cells. While we were writing this manuscript, Palucka et al published the results of a phase I clinical trial of a vaccine composed of DCs loaded with killed allogeneic melanoma cells which induced objective clinical responses and elicited MART 1 specific CD8 T cells in stage IV patients.

Thus, the use of a mi ture of apoptotic necrotic inhibitor Regorafenib allogeneic melanoma cell lines as a comple source of melanoma Ags for DCs cross presentation could be further e plored to validate and complement these findings in the clinical setting. The induction of apoptosis by different methods may pro duce a mi ture of apoptotic late apoptotic and or necrotic tumor cells that could provide different signals necessary for DCs maturation as well as for CTL priming. To investigate these points we produced and characterized human monocyte derived DCs and co incubated them with a mi ture of four melanoma cell lines, rendered ap