Nilsson Christina, Forsman Johanna, Hassan Zuzana, Abedi-Valugerdi Manuchehr, O'Connor Carmel, Concha Hernan, Jansson Monika, Hassan Moustapha
Laboratory of Hematology, Division of Hematology, Department of Medicine Karolinska University Hospital, Huddinge, Stockholm, Sweden.
Exp Hematol. 2005 Mar;33(3):380-7. doi: 10.1016/j.exphem.2004.12.003.
The present study was designed to investigate the influence of the administration sequence of busulphan (Bu) and cyclophosphamide (Cy) during conditioning regimen on myeloablative and immunosuppressive effects and on engraftment.
Female Balb/C mice were treated with either Bu-Cy or Cy-Bu (assigned order of administration). Bu was administered as 8.75 mg/kg/day x 4 and Cy as 100 mg/kg/day x 2. The control consisted of untreated animals. Bone marrow and spleen were harvested during the conditioning regimen and for up to 19 days after treatment. Colony-forming unit granulocyte macrophage assay was performed on marrow cells. Immunological analyses were performed using spleen cells. Liver status was determined using aspartate amino transferase (AST), alanine amino transferase (ALT), and bilirubin. Animals assigned for engraftment study were conditioned as above and transplanted using sca-1 cells from male Balb/C donors. Engraftment was followed using fluorescence in situ hybridization up to 30 days posttransplantation.
No significant difference in myeloablative effect was observed between treatments. Immunosuppressive activity expressed as CD3+/CD19+ and CD4+/CD8+ was also similar. Levels of cytokines interleukin 2, tumor necrosis factor alpha, and interferon gamma at the end of the conditioning regimen were lower in the Cy-Bu group, while liver enzymes were higher after the Bu-Cy regimen. Engraftment in bone marrow was reached faster within the first 20 days after conditioning with Cy-Bu compared to Bu-Cy. However, no difference in chimerism was observed at 30 days.
Cy-Bu treatment resulted in lower levels of cytokines, faster bone marrow engraftment, and lower values of liver enzymes compared to Bu-Cy regimen, which may benefit stem cell transplantation outcomes.
本研究旨在探讨预处理方案中白消安（Bu）和环磷酰胺（Cy）的给药顺序对清髓和免疫抑制作用以及植入的影响。
雌性Balb/C小鼠接受Bu-Cy或Cy-Bu（指定给药顺序）治疗。Bu的给药剂量为8.75mg/kg/天×4天，Cy为100mg/kg/天×2天。对照组为未治疗的动物。在预处理方案期间及治疗后长达19天收集骨髓和脾脏。对骨髓细胞进行集落形成单位粒细胞巨噬细胞测定。使用脾细胞进行免疫分析。使用天冬氨酸氨基转移酶（AST）、丙氨酸氨基转移酶（ALT）和胆红素测定肝脏状态。分配用于植入研究的动物按上述方法进行预处理，并使用来自雄性Balb/C供体的sca-1细胞进行移植。移植后使用荧光原位杂交监测植入情况直至30天。
各治疗组之间在清髓效果上未观察到显著差异。以CD3+/CD19+和CD4+/CD8+表示的免疫抑制活性也相似。预处理方案结束时，Cy-Bu组的细胞因子白细胞介素2、肿瘤坏死因子α和干扰素γ水平较低，而Bu-Cy方案后肝酶水平较高。与Bu-Cy相比，Cy-Bu预处理后第1个20天内骨髓植入更快。然而，在30天时嵌合率未观察到差异。
与Bu-Cy方案相比，Cy-Bu治疗导致细胞因子水平较低、骨髓植入更快且肝酶值较低，这可能有利于干细胞移植结果。
