地中海贫血患儿血清特异性PRA影响脐血造血干细胞增殖、分化能力的观察  

Effect of different PRA serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem cells /progenitor cells of cord blood

作　　者：翁文骏[1] 方建培[1] 吴燕峰[2] 肖露露[3] 徐宏贵[1] 包蓉[2] 魏菁[4] 许吕宏[1] 

机构地区：[1]中山大学附属第二医院儿科,广东广州510120 [2]中山大学附属第二医院脐血库,广东广州510120 [3]中山大学组织配型中心,广东广州510089 [4]中山大学附属第二医院医研中心,广东广州510120

出　　处：《中国病理生理杂志》2007年第11期2200-2204,共5页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No04009371);广东省卫生厅基金资助项目(NoA2005225);广东省科技厅重点引导资助项目(No2005B30701010)

摘　　要：目的:探讨反复输血地中海贫血患儿血清中特异性群体反应性抗体(PRA)对脐血造血干/祖细胞增殖、分化能力的影响。方法:采用1×105脐血单个核细胞(MNC)与实验血清(健康儿童AB血清50μL、PRA血清0μL、50μL、100μL)、补体联合孵育后半固体集落培养,倒置显微镜下观察并计数第7d、第14d总集落数和各种集落数。结果:脐血造血干/祖细胞受PRA血清作用后,在半固体集落培养第7d,总集落数、CFU-GM分别为A组88.20±9.41、79.00±11.39和B组88.60±9.12、79.20±10.44,显著高于C组20.60±7.39、15.20±4.66和D组4.00±2.05、1.40±0.51,P<0.01;其余各组的各种集落数两两比较,差异无显著(P>0.05)。A组与E组比较:两组的各种集落数比较均无显著差异(P>0.05)。在半固体集落培养第14d,总集落数、CFU-GM分别为A组216.00±31.10、117.40±24.80和B组213.20±31.06、116.00±19.75,显著高于C组97.80±14.43、32.80±8.10和D组31.40±13.41、8.40±4.30,P<0.01;第14dCFU-GEMM分别为A组45.60±8.51和B组42.60±7.03,显著高于C组20.80±6.96和D组7.80±6.06,P<0.05;第14dBFU-MK分别为A组12.80±4.42、B组11.00±2.74,显著高于D组1.00±0.55,P<0.05;B组第14dCFU-E17.20±4.03显著高于D组5.60±2.87,P<0.05。A组与E组比较:两组的各种集落数比较差异均无显著(P>0.05)。PRA血清量与各种集落数目之间的Kendall相关分析:PRA血清量与第7d的总集落数及CFU-GM、第14d的总集落数、CFU-GM、CFU-GEMM、BFU-E、BFU-MK呈负相关(tau-b分别为-0.793、-0.849、-0.808、-0.804、-0.645、-0.674、-0.624,P<0.01;与第14dCFU-MK呈负相关(tau-b为-0.466,P<0.05)。结论:特异性PRA对脐血造血干/祖细胞的增殖、分化能力具有抑制作用;在一定浓度下其抑制作用与PRA剂量有相关性:PRA剂量越大,抑制作用越强。AIM： To probe the effect of different panel reactive antibody （PRA） serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem/progenitor cell of cord blood. METHODS： 1×10^5 mononuclear cells （MNCs） isolated from umbilical cord blood were incubated with different PRA serum levels （0μL, 50μL, 100μL） respectively and complement, inoculated into the methylcellulose cultural system. The proliferation and differentiation potential of the hematopoietic stem/progenitor cell of cord blood by the colony formation assay were detected on day 7 and day 14, respectively. RESULTS： After culture of 7 days, the total colonies and CFU - GM were 88.20±9. 41,79.00±11.39 in group A and 88.60±9. 12, 79. 20±10. 44 in group B, which were signifi-cantly higher than those of 20.60±7. 39, 15.20±4.66 in group C and those of 4.00±2.05,1.40±0. 51 in group D （P〈 0. 01 ）. Meanwhile compared with group A and group E, no significant difference was found （P 〉0. 05）. After culture for 14 days, the total colonies and CFU - GM were 216. 00±31.10,117.40± 24. 80 in group A and 213.20 ±31.06,116. 00±19. 75 in group B, which were significantly higher than those of 97. 80± 14. 43,32. 80±8. 10 in group C and those of 31.40±13.41,8. 40±4. 30 in group D （P〈0.01）. The CFU - GEMMs were 45.60±8. 51 in group A and 42.60±7. 03 in group B, which were significantly higher than those of 20. 80±6. 96 in group C and those of 7. 80±6. 06 in group D （ P 〈 0.05 ）. The BFU - MK was 12. 80±4. 42 in group A and 11.00±2. 74 in group B respectively, which were significantly higher than that of 1.00±0.55 in group D （P〈0.05）. The CFU-E in group B 17.20±4.03 was significantly higher than that of 5.60±2.87 in group D （P 〈 0.05 ）. Meanwhile compared with group A and group E, no significant difference was found （ P 〉 0. 05）. By the Kendall test, there were negative correlations between the level of PRA serum and the total coloni

关 键 词：胎血 造血干细胞 地中海贫血 群体反应性抗体 

分 类 号：R363[医药卫生—病理学]