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ajt12756-sm-0001-SuppFig-S1.ppt72KFigure S1: Treatment with αIL-7R halts B cell development at an early stage. Bone marrow was collected from hCD52-Tg mice treated with PBS or αIL-7R and analyzed by FACS for the expression of B cell markers. The percentages of CD45R/220+ kappa- and kappa+ cells were reduced in mice treated with αIL-7R (0.4% vs 15.2%, and 0.2% vs 3.0%).
ajt12756-sm-0001-SuppFig-S2.ppt108KFigure S2: The combination of T- and B cell depletion does not promote long-term graft survival. (A) Spleen cells were collected from hCD52-Tg mice not treated (n = 3), treated with campath (n = 3), with αCD20 (n = 3), or with both drugs (n = 4) on day 7. Vital cells were counted and characterized by FACS. Shown are means ±SD of absolute numbers per spleen of CD3+ cells (white bars) and CD19+ cells (gray bars). Absolute numbers were calculated as product of numbers of spleen vital cells and percentage of CD3+ or CD19+ cells. (B) Fully mismatched B6 skin grafts were transplanted to hCD52-Tg mice under the cover of αCD20 alone (Δ, n = 6) or in combination with campath (□; n = 5). Lymphocyte antibody depletion could not prevent graft rejection in any mice; however treatment with αCD20 and campath significantly delayed rejection compared to αCD20 alone (MST 19 and 11 days, respectively, p = 0.0015).
ajt12756-sm-0001-SuppFig-S3.ppt114KFigure S3: Treatment with αIL-7R reduces the production of anti-donor MHC antibodies. Serum was collected from naïve hCD52-Tg mice (n; n = 2) or hCD52-Tg mice 3 weeks after the transplantation of B10 skin grafts under the cover of different treatments. Serum was then incubated with B10 thymocytes at progressive dilutions, and relative quantification of anti-donor MHC antibodies (mouse anti-mouse H-2b) was determined by FACS using a goat anti-mouse IgG as a secondary antibody. The level of anti-donor antibodies in mice treated with campath+αIL-7R, irrespective of graft acceptance (; n = 3) or rejection (Δ; n = 3), was lower than that in grafted mice treated with campath alone (□; n = 4) or with PBS (; n = 2) (p < 0.0001 for both comparisons). No difference in antibody titers was found between mice treated with campath+αIL-7R with a viable graft (n = 3) compared to those that had rejected their graft (n = 3) (p > 0.05).

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