The experiment was designed to have 80% power to detect a 15 day difference in survival
The experiment was designed to have 80% power to detect a 15 day difference in survival. cell transplantation is performed for high risk acute lymphoblastic leukemia (ALL). Relapse of ALL remains the most common cause of treatment failure after transplant. While augmentation of graft versus leukemia effects by withdrawal of immunosuppression and donor lymphocyte infusion is definitely often effective in treating early relapse of chronic myelogenous leukemia after transplant, these maneuvers are hardly ever successful in treatment of ALL (1, 2). The mechanisms of the relative ineffectiveness of donor lymphocyte infusion and graft versus leukemia activity in ALL are not fully known. Some of the options include rapid development of ALL cells in vivo and poor immunogenicity of ALL cells compared to CML cells. Prior work in our laboratory has shown that administration of cellular leukemia vaccines to Ro 90-7501 allogeneic transplant recipients can increase graft versus leukemia effects without substantial raises in graft Ro 90-7501 versus sponsor disease (3, 4). We have also observed that vaccination at the time of allogeneic lymphocyte infusion can result in a significant development of antigen specific T cells in vivo (5). Based Ro 90-7501 on these findings we hypothesized that vaccination coupled with donor lymphocyte infusion might create more effective control of ALL after transplant. The reasoning for this was that active vaccination would provide more effective antigen demonstration of antigens present on ALL cells and that the clonal development of leukemia reactive T cells might be more effective in controlling ALL populations that have a rapid development rate. To address this immunobiological query we employed a well characterized murine model of MHC-matched, multiple small histocompatibility antigen mismatched transplantation (6), and novel murine pre-B acute lymphoblastic leukemia cell lines driven by common human being mutations (bcr/abl fusion genes and Ink/ARF locus deletions) (7). While many of the small histocompatibility antigens in this system are known at a genetic and peptide level (6), antigens relatively selectively indicated within the leukemia cells are not. To address this methodological limitation we exploited sex-mismatches since male HY antigens are known at a genetic and peptide level. By using ALL cells derived from males and using female donors and recipients we were able to use the male HY antigens as models for leukemia restricted antigens (8). We discovered that while concurrent vaccination Ro 90-7501 did increase the activity of donor T cells that identified HY antigens within the leukemias and did have a short term impact on leukemia development, significant survival advantages were not seen by the addition of vaccination to donor lymphocyte infusion. Once we investigated the mechanisms of relapse and immune control of ALL with this model we discovered that long term survival after allogeneic transplant and ALL challenge were associated with moderate T reactions, and remarkably, B cell reactions to leukemia cells. MATERIALS AND METHODS Mice C3.SW mice (Jackson) were transplant donors and C57BL/6 mice (National Tumor Institute, Frederick, MD) were recipients. The mice are MHC antigen matched (H2b) but small histocompatibility antigen (mHA) mismatched at many loci (H1, H3, H7, H8, H9, H13). C3.SW are H2b and were derived from Rabbit Polyclonal to LAT an 11 generation back mix of C3H against a non-inbred H2b donor strain (9). Cell lines NSTY pre-B acute lymphoblastic leukemias were generated from main marrow cells from INK4A/ARF null mice transduced having a retroviral vector encoding the human being p210 Ro 90-7501 bcr/abl cDNA (7, 10). The MSCV-BCR/ABL-IRES-GFP vector was kindly provided by Dr. Richard Vehicle Etten. The neo gene was removed from this vector by digestion with Nco I and Cla I, and the YFP gene was put by standard cloning process to yield the MSCV-Nup98/HoxA9-YFP vector used in the present study. Retroviral vector plasmids were transfected into phoenix-eco cells (ATCC) using lipofectamine 2000 per manufacturers instructions (10 micrograms DNA per 100,000 cells inside a six-well tissue tradition dish). At 36 hours post-transfection, viral supernatants were collected, filtered, and stored at ?80 degrees centigrade. Retrovirus treated marrow cells (2 105) were infused.