地中海贫血患儿血清特异性群体反应性抗体对脐血造血干细胞增殖、分化能力的影响(英文)

Thalassemic serum panel reaction antibody inhibits proliferation and differentiation of cord blood hematopoetic stem cells in children patients

背景：已有报道证实群体反应性抗体在实体器官移植受者排斥反应中起着重要作用，并且与植入物功能衰竭呈正相关。目的：实验拟观察反复输血地中海贫血患儿血清中特异性群体反应性抗体对脐血造血干，祖细胞增殖、分化能力的影响。设计、时间及地点：体外细胞学实验，于2006—01／2007—08在中山大学附属第二医院医学研究中心完成。材料：5份健康足月产妇脐带血，每份80～100mL。反复输血地中海贫血患儿的群体反应性抗体血清样本。5份健康儿童AB分型血清样本。6份阳性抗凝静脉血样本10mL。方法：采用聚蔗糖泛影葡胺梯度离心法分离脐带血单个核细胞。将1×10^5脐血单个核细胞与分别与健康儿童AB血清50uL、0，50，100uL不同浓度群体反应性抗体血清混合后，再将上述细胞混合物与兔补体共同孵育进行半固体集落培养。主要观察指标：倒置显微镜下观察并计数共培养第7，14天总集落数和各种集落数。结果：脐血造血干，祖细胞受群体反应性抗体血清作用后，其半固体集落培养的总集落数以及各种集落数均有不同程度的下降，其中总集落数和粒单系集落形成单位的下降数，经统计学处理差异具有显著性意义（P〈0．01）：群体反应性抗体血清量与总集落、粒单系集落形成单位、粒红单核巨核系集落形成单位、红系爆式集落形成单位、巨核系爆式集落形成单位、巨核系集落形成单位之间的肯德尔相关分析呈负相关，经统计学处理差异有显著性意义（P〈0．05）。结论：特异性群体反应性抗体对脐血造血干／祖细胞的增殖、分化能力具有抑制作用，并且抑制作用与群体反应性抗体剂量相关：群体反应性抗体剂量越大，抑制作用越强。

BACKGROUND： Panel reaction antibody （PRA） plays an important role in rejection of recipients undergoing solid organ transplantation, which has a positive effect on nonfunction of implant. OBJECTIVE： To evaluate the effect of thalassemic serum-specific PRA on the proliferation and differentiation of umbilical cord blood hematopoetic stem/progenitor cells （HSC/HPCs） in children patients with thalassemia. DESIGN, TIME AND SETTING： The in vitro cytology experiment was performed at the Experimental Research Center, Second Affiliated Hospital, Zhongshan University from January 2006 to August 2007. MATERIALS： Five samples of umbilical cord blood from healthy full-term birth puerperants （each 80-100 mL） were used in this study. PRA serum samples of children patients with thalassemia after repetitive blood transfusion, five samples of AB blood grouping serum, and six samples of positive anticoagulation vein blood （10 mL） were used in the study. METHODS： Mononuclear cells were harvested from umbilical cord blood by Ficoll-Hypaque gradient centrifugation. 1 × 10^5 mononuclear ceils from umbilical cord blood were incubated with different levels of experimental or AB control serum （0, 50,100 u L） from healthy children. The mixture mentioned above was incubated with rabbit complement for semisolid colony culture. MAIN OUTCOME MEASURES： Colony-forming units （CFU） were counted and observed after 7 days and 14 days of culture under an inverted microscope. RESULTS： After incubation with HSC/HPCs PRA serum, total number of CFUs and varied CFUs decreased to different extents, of which the total number of CFUs and CFU- granulocyte-macrophages （CFU-GM） had significant differences （P 〈 0.01）. Moreover, there were negative correlations between different levels of serum PRA and the followings： number of total colonies, CFU- GM, CFU- granulocyte-erythrocyte-monocyte-megakaryocytes, CFU-erythroids, burst forming unit-megakaryocytes, and CFU-megakaryocytes （P 〈 0.05）. CONCLUSION： The thalassemic serum PRA has an apparent inhibitory effect on the proliferation and differentiation of cord blood HSC/HPCs in vitro, an effect that may be pronounced with increasing serum PRA.