1. Donor antigen-presenting cells regulate T-cell expansion and antitumor activity after allogeneic bone marrow transplantation 
Jian-Ming Li, Edmund K. Waller 
Biology of Blood and Marrow Transplantation 
Volume 10, Issue 8, Pages (August 2004)
DOI: /j.bbmt

2. Figure 1
Phenotypes of DCs and DC precursor subsets in freshly isolated BM cells and splenocytes. Cells from BM (A-D) and spleen (E and F) of C57BL/6 mice were stained with CD11c, B220, CD11b, CD4, CD8, and a lineage (Lin) cocktail consisting of anti-CD3, CD5, CD19, DX5, surface immunoglobulin M, and TER-119 monoclonal antibodies. A, Electronic gates were placed by using forward and side scatter properties of large lymphocytes and monocytes in BM. B, Electronic gates were placed around the subpopulation of cells defined in panel A limited to those cells lacking lineage marker expression and expressing low to high levels of CD11c. C and E, The population of cells included in the gates defined in panels A and B was further subdivided on the basis of expression patterns of MHC class II (I-Ab) and CD11b. D and F, The population of cells included in the gates defined in panels A and B was subdivided on the basis of expression patterns of CD11b and CD4.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )

3. Figure 2
GVHD clinical scores of recipients of a combination of unmanipulated or CD11b-depleted BM and graded doses of donor splenocytes. Clinical signs of GVHD (weight loss, posture, activity, fur texture, and skin integrity; maximum score, 10) were assessed once or twice per week up to day 100 according to the procedure described in Materials and Methods. Mean values for the clinical GVHD score at each time point were determined in groups of mice that received unmanipulated BM grafts (A) or CD11b-depleted BM grafts (B) plus no added splenocytes (○), 0.5 × 106 splenocytes (▵), 1 × 106 splenocytes (▴), 5 × 106 splenocytes (□), and 10 × 106 splenocytes (♢). The data shown represent the overall mean values ± SD for all clinical GVHD scores at each of 17 time points for each group and are representative of similar results seen in 3 separate experiments. ∗P < .05 comparing the GVHD score between recipients of an unmanipulated BM graft versus CD11b-depleted BM with same dose of splenocytes.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )

4. Figure 3
Expansion of donor-derived T cells after transplantation was enhanced by CD11b depletion of the BM graft. CD45.2+ B10.BR recipient mice were lethally irradiated with 11 Gy on day −2 and underwent transplantation with 5 × 106 unmanipulated CD45.1+CD90.2+ C57BL/6 BM alone (♢) or 1 × 106 CD45.1+CD90.1+ C57BL/6 donor splenocytes combined with unmanipulated BM (□), CD11b [+/−] BM (▵), CD11b [−] BM (○), or CD11b [+] BM (♦). Complete blood counts and flow cytometry for T-cell subsets were performed on samples of peripheral blood obtained on day +30, day +60, and day +105 after BMT. The transplantation experiment was repeated at least 3 times for each condition, and 5 to 10 mice underwent transplantation for each condition in each experiment. A, The mean ± SD of the absolute number of CD45.1+CD90.2+ donor BM-derived T cells per microliter of peripheral blood. B, The mean ± SD of CD45.1+CD90.1+ donor spleen graft-derived T cells per microliter of peripheral blood. C, The level of donor spleen-derived T cells in peripheral blood on day +60 after transplantation was determined in recipients of unmanipulated BM grafts (filled bar) or CD11b [−] BM grafts (open bar) with 0.3 × 106 purified donor spleen T cells. D, The correlation between the content of CD11b+ DCs and DC precursors (♢) and CD11b− DCs and DC precursors (•). The mean value of spleen-derived T cells per microliter of peripheral blood was determined from multiple experiments with different numbers of BM cells. E, Mean values ± SD of IFN-γ were determined by enzyme-linked immunosorbent assay from day +30 serum samples of peripheral blood obtained from 5 mice per group of B10.BR mice lethally irradiated and transplanted with a combination of 5 × 106 unmanipulated or CD11b-depleted C57BL/6 BM plus 0.3 × 106 MACS-purified C57BL/6 donor T cells. ∗P <. 05; ∗∗∗P < .001 comparing the mean values from recipients of CD11b-depleted BM/splenocyte grafts with the recipients of same number of unmanipulated BM/splenocytes.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )

5. Figure 4
GVL activity of BM and spleen grafts against LBRM in a day 75 challenge model was enhanced by CD11b depletion of the BM graft in C57BL/6→B10.BR transplants. B10BR recipient mice were lethally irradiated (11 Gy) on day −2 and received various combinations of BM and splenocytes from C57BL/6 donors on day 0. The GVL effect of the allogeneic transplant was assessed by administration of a lethal dose of 3 × 106 LBRM cells on day +75 after transplantation. The content of CD11b+ cells in the BM graft varied among unmanipulated BM (□), CD11b [−] BM (○), CD11b [+/−] BM (▵), and CD11b [+] BM (♢). Each treatment group contained 15 or 20 mice pooled from 3 or 4 experiments. A, Survival of mice transplanted with BM alone after LBRM challenge on day +75. B, Survival of mice transplanted with CD11b-depleted BM and 1 × 106 splenocytes with LBRM administered on day +75 after transplantation. C, Representative GVHD histopathology of liver from a mouse transplanted with CD11b-depleted BM and 1 × 106 splenocytes, with LBRM administered on day +75 after transplantation, that was killed on day 220 after transplantation. D, Representative liver histopathology from a mouse transplanted with unmanipulated BM and 1 × 106 splenocytes, with LBRM administered on day +75 after transplantation, that was killed on day 120 after transplantation.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )

6. Figure 5
GVL activity of BM and spleen grafts against LBRM in a day −1 challenge model was enhanced by partial CD11b depletion of the BM graft in C57BL/6→B10.BR transplants. B10.BR recipient mice were lethally irradiated (11 Gy) on day −2 and received various combinations of BM and splenocytes from C57BL/6 donors on day 0. The GVL effect of the allogeneic transplant was assessed by administration of a lethal dose of 3 × 106 LBRM cells 1 day before transplantation. The content of CD11b+ cells in the BM graft varied among unmanipulated BM (□), CD11b [−] BM (○), CD11b [+/−] BM (▵), and CD11b [+] BM (♢). Each treatment group contained 15 or 20 mice pooled from 3 to 4 experiments. A, Survival of mice transplanted with LBRM on day −1 with 5 × 106 BM alone (■), 5 × 106 BM with 1 × 106 splenocytes (▴), 5 × 106 BM with 5 × 106 splenocytes (•), and 5 × 106 BM with 10 × 106 splenocytes (♦) on day 0. A control group received 1 × 106 BM with 1 × 106 splenocytes without LBRM (filled cross, dashed line). B, Survival of mice transplanted with BM alone and LBRM on day −1. C, Survival of mice transplanted with BM grafts and 1 × 106 splenocytes, and LBRM on day −1.
Biology of Blood and Marrow Transplantation  , DOI: ( /j.bbmt )