A CD4 Domain 1 CC′ Loop Peptide Analogue Enhances Engraftment in a Murine Model of Bone Marrow Transplantation with Sublethal Conditioning Gabor Varadi,

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Presentation transcript:

A CD4 Domain 1 CC′ Loop Peptide Analogue Enhances Engraftment in a Murine Model of Bone Marrow Transplantation with Sublethal Conditioning Gabor Varadi, Thea M. Friedman, Robert Korngold Biology of Blood and Marrow Transplantation Volume 11, Issue 12, Pages 979-987 (December 2005) DOI: 10.1016/j.bbmt.2005.08.038 Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 1 Radiation dose titration effect on donor engraftment in the MHC class II–mismatched bm12 → B6-CD45.1 strain combination. B6-CD45.1 mice (n = 3-5) were exposed to irradiation with the indicated dosage and underwent transplantation with allogeneic bm12 ATBM (107 cells). Splenocytes from recipients were analyzed for donor chimerism on day 30 after BMT by flow cytometry with FITC–anti-CD45.1 (expressed by recipient) and PE–anti-CD45.2 (expressed by donor) mAb. The data are presented as the mean percentage of chimerism ± SD and are from 1 representative experiment of 2 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 2 Effect of 802-2 peptide on splenic donor mononuclear cell engraftment 30 days after BMT. B6-CD45.1 mice (n = 3-5) were exposed to 800 cGy of irradiation and underwent transplantation with allogeneic bm12 ATBM (107 cells). Recipients were treated on day 0 with PBS, 802-2 peptide (0.5 mg), or anti-CD4 mAb. Mononuclear cells from recipient spleens were analyzed for donor/recipient chimerism on day 30 after BMT by flow cytometry with FITC–anti-CD45.1 and PE–anti-CD45.2 mAb. The donor expressions in the 802-2 peptide (76.5%) and anti-CD4 mAb (80.1%) treatment groups were significantly higher than in the PBS control group (38.3%; P < .001). The data (mean ± SD) are from 1 representative experiment of 2 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 3 Splenic donor mononuclear cell engraftment 72 days after BMT. B6-CD45.1 mice underwent transplantation with bm12 ATBM and were treated as described in Figure 2. Mononuclear cells from recipient spleens (n = 3-5) were analyzed for donor chimerism on day 72 after BMT. The donor expressions in the 802-2 peptide (73.5%) and anti-CD4 mAb (75.0%) treatment groups were significantly higher than in the PBS control group (38.3%; P < .03). The data (mean ± SD) represent the mean of 2 experiments. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 4 Donor CD4+ and CD8+ T-cell reconstitution in spleens 30 days after BMT. B6-CD45.1 mice underwent transplantation with bm12 ATBM and were treated as described in Figure 2. CD4+ and CD8+ T cells from recipient spleens (n = 3-5) were analyzed for donor chimerism and T-cell phenotype on day 30 after BMT. A, The donor expressions in the 802-2 peptide (56%) and anti-CD4 mAb (57.8%) treatment groups were significantly different from that of the PBS control group (36.0%; P < .002). B, The donor CD8+ T-cell values in the 802-2 peptide (59%) and anti-CD4 mAb (54.4%) treatment groups were marginally significantly different from that of the PBS control group (43.4%; P ≤ .09). The data (mean ± SD) are from 1 representative experiment of 2 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 5 Donor CD4+ and CD8+ T-cell reconstitution in lymph nodes 72 days after BMT. B6-CD45.1 mice underwent transplantation with bm12 ATBM and were treated as described in Figure 2. CD4+ and CD8+ T cells from recipient (n = 3-5) lymph nodes were analyzed for donor chimerism on day 72 after BMT by flow cytometry. A, The donor-derived CD4+ T-cell values in the 802-2 peptide (81%) and anti-CD4 mAb treatment (76%) groups were significantly higher than in the PBS control group (61%; P ≤ .04). B, Donor CD8+ T-cell chimerism was also significantly increased (P ≤ .002), from 57.6% in the PBS group to 81.6% and 74.4% in the 802-2–treated and anti-CD4–treated groups, respectively. The data (mean ± SD) are from 1 representative experiment of 2 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 6 Effect of 802-2 peptide on CFU-GM colony formation. B6-CD45.1 mice underwent transplantation with bm12 ATBM and were treated as described in Figure 2. On day 5 after BMT, splenocytes were pooled from 3 B6-CD45.1 mice per treatment group, plated in triplicate in semisolid methylcellulose media, and cultured for 6 days. Colonies that contained >30 CFU-GM cells were scored as positive. Increased numbers of colonies were formed in splenocyte cultures from the 802-2 peptide–treated (2.4-fold) and the anti-CD4 mAb–treated (4.1-fold) groups in comparison to the PBS control group (P ≤ .001). The results are from 1 representative experiment of 3 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions

Figure 7 Effect of 802-2 peptide on donor chimerism in presensitized recipients. B6-CD45.1 mice were presensitized (IP) with 2 × 107 bm12 splenocytes 14 days before irradiation exposure (800 cGy) and transplantation with bm12 ATBM (107 cells). Recipients were treated daily from day 0 to 3 with PBS, 802-2 peptide (0.5 mg), or anti-CD4 mAb. Donor chimerism was analyzed 30 days after BMT by flow cytometry with FITC–anti-CD45.1 and PE–anti-CD45.2 mAb. This treatment schedule caused a significant increase in donor chimerism in the 802-2–treated group (62%) compared with the PBS-treated control mice (15%; P ≤ .007). The data (mean ± SD) are from 1 representative experiment (n = 5 mice per group) of 2 performed. Biology of Blood and Marrow Transplantation 2005 11, 979-987DOI: (10.1016/j.bbmt.2005.08.038) Copyright © 2005 American Society for Blood and Marrow Transplantation Terms and Conditions