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Blood, Vol. 109, Issue 10, 4564-4574, May 15, 2007
Factors governing the activation of adoptively transferred donor T cells infused after allogeneic bone marrow transplantation in the mouse
Blood Durakovic et al.
109: 4564
Supplemental materials for: Durakovic et al, Vol 109, Issue 10, 4564-4574
Files in this Data Supplement:
- Figure S1. Effects of DLI administration from naive and presensitized donors on CD11c+ DC and blood chimerism in 8-week old mixed chimeras (PDF, 18 KB) -
(A-B) Analysis of CD11c+ DC chimerism in spleen and skin. 8-week-old B10.D2 + BALB/c-CD45.1→BALB/c-CD45.1 mixed chimeras received 2 × 107 splenocytes from either naive (black boxes) or presensitized B10.D2-Thy1.1 mice (white triangles). An additional group of B10.D2→BALB/c full chimeras received DLI from naive donor (black diamonds). Spleens (SPL) of the chimeras (n=3/group) were retrieved, digested with collagenase D, and analyzed by flow cytometry using fluorochrome-conjugated anti-CD11c– and anti-CD45.1–specific antibodies. Skin-derived CD11c+ DCs were obtained from the epidermal sheets after 2 days of in vitro culture in the presence of GM-CSF and TNF- . Emigrating live (PI−) cells were analyzed for expression of CD11c, CD45.1 (host), and CD45.2 (donor) by 4-color flow-cytometry (n=3 chimera/group/time point; *# P < .05, black squares versus black diamonds; and white triangles versus black diamonds). (C) DLI from intraperitoneal- and skin-presensitized donors are ineffective in inducing LH-GVH response potent enough to induce conversion from mixed to full donor chimerism in MHC-matched chimeras. Presensitized B10.D2 donors were generated by intraperitoneal administration of BALB/c splenocytes or by placing the BALB/c skin grafts on B10.D2 mice. Four weeks after intraperitoneal priming or at the time of skin graft rejection, spleen cells were collected and given as a DLI to the 4-week-old B10.D2 + BALB/c→B10.D2 mixed chimeras. Changes in donor CD4+ T-cell chimerism were determined by serial tail bleeding as described in Figure 1.
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