Akpinar Edip, Craighead Nancy, Smoot Douglas, Hale Douglas A
Transplantation Branch, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Transplantation. 2004 Aug 15;78(3):383-91. doi: 10.1097/01.tp.0000128833.38229.4f.
The infusion of donor bone marrow (BM) into mice conditioned with antilymphocyte serum (ALS) and sirolimus (Sir) prolongs skin allograft survival and produces chimerism. This study identifies the BM cell(s) responsible for this effect and determines whether enrichment for these cells will improve efficacy.
Skin grafts from BALB/C mice were transplanted into C57BL/6 or C57BL/10 recipients by using ALS, Sir, and BM (or fractions). BM was fractionated by using immunomagnetic beads. Flow cytometry was used for phenotyping and detecting chimerism.
The median graft survival in mice receiving 25 million BM cells was 61 days. Infusion of BM depleted of cells expressing CD19, CD3, CD11c, and c-kit had no effect on median graft survival, whereas infusion of fractions enriched for those cells resulted in median graft survival of 38, 48, 28, and 83 days, respectively. The administration of higher doses (4 x 10(6) and 8x10(6)) of fractions enriched for c-kit resulted in median graft survival of 124 and 197 days, respectively, without chimerism. This favorably compared with mice receiving 150 million BM cells that demonstrated transient mixed chimerism and a median graft survival of 190 days. The majority of cells in the c-kit+-enriched fraction expressed lineage markers. Removal of lineage positive cells from BM before infusion shortened median graft survival (90 days), indicating that the c-kit+ lin+ population is largely responsible for prolongation of graft survival.
Cells enriched for C-kit+lin+ constitute approximately 5% of murine BM cells and are more potent than whole BM at prolonging skin allograft survival in mice treated with ALS and Sir.
将供体骨髓（BM）输注到用抗淋巴细胞血清（ALS）和西罗莫司（Sir）预处理的小鼠体内可延长皮肤同种异体移植的存活时间并产生嵌合体。本研究确定了造成这种效应的BM细胞，并确定富集这些细胞是否会提高疗效。
通过使用ALS、Sir和BM（或组分），将BALB/C小鼠的皮肤移植到C57BL/6或C57BL/10受体小鼠体内。使用免疫磁珠对BM进行分级分离。流式细胞术用于表型分析和检测嵌合体。
接受2500万个BM细胞的小鼠的移植中位存活时间为61天。输注去除表达CD19、CD3、CD11c和c-kit的细胞的BM对移植中位存活时间没有影响，而输注富集这些细胞的组分导致移植中位存活时间分别为38天、48天、28天和83天。给予更高剂量（4×10⁶和8×10⁶）的富集c-kit的组分导致移植中位存活时间分别为124天和197天，且无嵌合体。这与接受1.5亿个BM细胞的小鼠相比具有优势，后者表现出短暂的混合嵌合体，移植中位存活时间为190天。富集c-kit⁺组分中的大多数细胞表达谱系标志物。输注前从BM中去除谱系阳性细胞会缩短移植中位存活时间（90天），表明c-kit⁺lin⁺群体在很大程度上负责移植存活时间的延长。
富集的c-kit⁺lin⁺细胞约占小鼠BM细胞的5%，在延长接受ALS和Sir治疗的小鼠皮肤同种异体移植存活时间方面比全BM更有效。
