Tumor regression by anti-CD40 and interleukin-2: role of CD40 in hematopoietic cells and organ-specific effects Lisbeth A. Welniak, Lynnette Shorts,

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Tumor regression by anti-CD40 and interleukin-2: role of CD40 in hematopoietic cells and organ-specific effects Lisbeth A. Welniak, Lynnette Shorts, Jeff Subleski, Bruce R. Blazar, Robert H. Wiltrout, William J. Murphy Biology of Blood and Marrow Transplantation Volume 10, Issue 8, Pages 534-539 (August 2004) DOI: 10.1016/j.bbmt.2004.05.006

Figure 1 Renca orthotopic tumor model and treatment schema. Biology of Blood and Marrow Transplantation 2004 10, 534-539DOI: (10.1016/j.bbmt.2004.05.006)

Figure 2 CD40 expression in BMC chimeras. A, One month after bone marrow transplantation, 2 mice from CD40+/+ or CD40−/− bone marrow chimeras were analyzed for CD40 expression on spleen cells to confirm the absence of CD40 in CD40−/− bone marrow chimeras. Representative histograms from 1 experiment are presented. B, Three mice from CD40+/+ or CD40−/− bone marrow chimeras were analyzed for CD40 expression on spleen cells to confirm the absence of CD40 in the dendritic cell population (CDIIc+/MHC class II+) in CD40−/− bone marrow chimeras 2 days after immunotherapy in tumor-bearing animals. Representative histograms for CD40 expression in CD11c+ MHC class II+ gated cell populations are presented. KO, knockout; WT, wild type. Biology of Blood and Marrow Transplantation 2004 10, 534-539DOI: (10.1016/j.bbmt.2004.05.006)

Figure 3 Antitumor effects of anti-CD40 and IL-2 on radiation BMC chimeras. CD40 chimeric mice were engineered by transplanting bone marrow cells from CD40+/+ or CD40−/− mice into myeloablated CD40+/+ recipients. Mice received intrarenal injections of Renca followed 11 days later by unilateral nephrectomy and the standard course of treatment with anti-CD40 and IL-2 (triangle symbols) or rat immunoglobulin G and vehicle controls (square symbols) for 12 days (as described in Materials and Methods; n = 13 mice per group). CD40+/+ bone marrow chimeras (▴) that received anti-CD40 and IL-2 therapy, but not CD40+/+ bone marrow chimeras that received control injections (■) or CD40−/− bone marrow chimeras (open symbols), responded with a significant increase in survival (log-rank test; P < .005). The results are the combination of 2 independent experiments. KO, knockout; WT, wild type; Ig, immunoglobulin. Biology of Blood and Marrow Transplantation 2004 10, 534-539DOI: (10.1016/j.bbmt.2004.05.006)

Figure 4 Effect of anti-CD40 and IL-2 on CD40+ splenic dendritic cell expansion in radiation BMC chimeras. Splenic dendritic cells were identified by MHC class II and CD11c coexpression. CD40 expression was determined for this population of cells from CD40+/+ and CD40−/− bone marrow chimeras that received either rat immunoglobulin G and vehicle control or anti-CD40 and IL-2. KO, knockout; WT, wild type; Ig, immunoglobulin. Biology of Blood and Marrow Transplantation 2004 10, 534-539DOI: (10.1016/j.bbmt.2004.05.006)