Early Vaccination with Tumor Lysate-Pulsed Dendritic Cells after Allogeneic Bone Marrow Transplantation Has Antitumor Effects Jeffrey S. Moyer, Gabriel.

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Early Vaccination with Tumor Lysate-Pulsed Dendritic Cells after Allogeneic Bone Marrow Transplantation Has Antitumor Effects Jeffrey S. Moyer, Gabriel Maine, James J. Mulé Biology of Blood and Marrow Transplantation Volume 12, Issue 10, Pages 1010-1019 (October 2006) DOI: 10.1016/j.bbmt.2006.06.009 Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 TL-pulsed DC vaccination alone does not have a significant antitumor effect . A, 7-day subcutaneous B16 tumors (5 × 104) in BL6 mice were treated with 1 × 106 BL6 DC delivered subcutaneously to the contralateral flank on days 7 and 11 after tumor inoculation. Mice were treated with PBS (circles), unpulsed DC (diamonds), Trp2 (180-188)-pulsed DC (triangles), or TL-pulsed DC (squares). There were no statistically significant differences between treatment groups (P > .05). B, Mice were immunized with 3 weekly DC vaccinations (1 × 106) and then challenged on day 4 after the final vaccination with 1 × 105 B16 tumor cells. Mice were vaccinated with PBS (diamonds), unpulsed DC (triangles), or TL-pulsed DC (squares). There were no statistically significant differences between treatment groups (P > .05). Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Early treatment after allogeneic (allo) BMT with TL-pulsed DCs has antitumor effects. Six-day subcutaneous B16 tumors (1 × 105) were treated with 1000 rads split-dose TBI and allogeneic BMT (1 × 107 cells). Three weekly doses of 1 × 106 DCs were delivered subcutaneously in the flank contralateral to the tumor on days 3, 10, and 17 after BMT. Mice were treated with allogeneic BMT alone (solid squares), unpulsed DC (open squares), Trp2 (180-188)-pulsed DC (triangles), or TL-pulsed DC (circles). *P < .0001. No animals were found to be tumor free after TL-pulsed DC vaccinations. This experiment was performed twice with consistent results. Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Early immunization after allogeneic BMT does not confer protection from tumor challenge. B6 mice were treated with 1000 rads split-dose TBI and allogeneic BMT (1 × 107 cells). Mice were immunized with 3 weekly DC vaccinations (1 × 106) starting on day 3 after BMT and then challenged on day 7 (day 24 after BMT) after the final vaccination with 1 × 105 B16 tumor cells. Mice were vaccinated with PBS (solid squares), unpulsed DC (open squares), or TL-pulsed DC (circles). There were no statistically significant differences between treatment groups (P > .05). Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Donor-derived (Ly 9.1+) T cell engraftment 24 days after BMT in the treatment model. Splenocytes from recipient mice treated with the various DC regimens were analyzed for the presence of the Ly 9.1 marker present in 100% of donor T cells. Results are from pooled samples from 2 mice. *P < .05 versus BMT alone. Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 Donor-derived (Ly 9.1+) T cell engraftment 24 and 38 days after BMT in the protection model. Splenocytes from recipient mice treated with the various DC regimens were analyzed for the presence of the Ly 9.1 marker present in 100% of donor T cells. Results are from pooled samples from 2 mice. *P < .05 versus BMT alone. Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 6 Absence of significant cytolytic immune response 24 days after BMT in the treatment model. Splenocytes were harvested and pooled from 3 mice treated with allogeneic (allo) BMT alone or with allogeneic BMT and TL-pulsed DCs. T cells (2 × 105) were stimulated for 5 days with 2 × 104 irradiated, IFN-γ-treated B16 cells or with 2 × 104 irradiated Trp2 (180-188)-pulsed DCs or TL-pulsed DCs. Lymphocyte-activated killer (LAK) cells were generated by culturing splenocytes for 5 days in 6000 IU/mL IL-2.51Cr-labeled B16 cells were used as targets in the chromium release assay at an effector:target ratio of 10:1. Biology of Blood and Marrow Transplantation 2006 12, 1010-1019DOI: (10.1016/j.bbmt.2006.06.009) Copyright © 2006 American Society for Blood and Marrow Transplantation Terms and Conditions