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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1×105脐血单个核细胞与分别与健康儿童AB血清50μL、0,50,100μL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: It has been reported that group-reactive antibodies play an important role in the rejection of solid organ transplant recipients and are positively correlated with implant failure. OBJECTIVE: To observe the effect of specific antibody in serum of repeated transfused children with thalassemia on the proliferation and differentiation of cord blood hematopoietic stem / progenitor cells. DESIGN, TIME AND SETTING: In vitro cytology experiments were performed at the Medical Research Center of the Second Affiliated Hospital of Sun Yat-sen University from January 2006 to August 2007. Materials: 5 healthy full-term maternal umbilical cord blood, each 80 ~ 100mL. Serum samples from patients with repeated transfusion of thalassemia major. 5 healthy children AB typing serum samples. 6 positive anticoagulation venous blood samples 10mL. Methods: Umbilical cord blood mononuclear cells were isolated by Ficoll - diacetate gradient centrifugation. 1 × 10 5 cord blood mononuclear cells were mixed with 50 μL AB serum, 0, 50, 100 μL serum of different concentrations of group AB from healthy children respectively, and then the above cell mixture was incubated with rabbit complement for semi-solid colony culture. MAIN OUTCOME MEASURES: The total number of colonies and the number of colonies on the 7th and 14th day of co-culture were observed and counted under an inverted microscope. Results: After the cord blood hematopoietic stem / progenitor cells were affected by serum of group reactive antibody, the total number of colonies and the number of all kinds of colonies in the semi-solid colony culture decreased with different degrees. The total number of colony and single colony forming units (P <0.01). The serum antibody levels of colorectal reactive antibody and total colony, single colony forming unit of granulocyte, mononuclear megakaryocyte colony forming unit of granulocyte, erythrocytic burst (P <0.05). The Kendall correlation analysis between the colony forming units, the megakaryocytic burst forming units and the megakaryocyte colony forming units was negatively correlated. The difference was statistically significant (P <0.05). CONCLUSIONS: The specific group reactive antibody can inhibit the proliferation and differentiation of cord blood hematopoietic stem / progenitor cells, and the inhibitory effect is related to the dose of colony-reactive antibody. The larger the dose of colorectal antibody, the stronger the inhibition.