微卫星估价异基因外周血干细胞移植后嵌合状态

Evaluating hematopoietic chimerism following allogeneic stem cell transplantation with microsatellites.

目的 用检测 5个四核苷酸重复序列了解异基因干细胞移植后嵌合状态以及预后的关系。方法 PCR扩增12 0份正常人和 10对移植供受者基因组DNA ,据所得片段大小确定 5个微卫星在人群中的多态性及移植后嵌合状态。结果 各微卫星均发现 7个以上等位基因 (7～ 12 )多态性 ,杂合度仅CSF1PO低于 70 %。移植患者中完全嵌合状态 (CC) 8例 ,1例死于GVHD ;混合嵌合状态 (MC) 1例 ,死亡 ;受者型 1例 ,死亡。结论 5个四核苷酸重复序列点多态性可成功检测大多数嵌合状态。

Objective: To study the feasibility of five selected tetranucleotide repeat sequences, detect chimerism of patients who underwent allogeneic peripheral blood stem cell transplantation(allo-PBSCT)and the correlation between chimerism status and prognosis of patients.Methods: Amplify genomic DNA of umbilical cord blood from 120 normal donors and 10 pairs of transplant siblings by polymerase chain reaction(PCR).PCR products were electrophoresed on PAGE gels.For each locus,fragments of different size were collected to analyze polymorphism.Genotypes of each siblings were analyzed at every locus to determine chimerism.For sex-mismatched transplantation,the amelogenin locus was analyzed in which discriminates between X and Y chromosome.Correlation between chimerism and prognosis was observed.Results: Three symbols including the number of alleles and genotypes,heterozygosity were used to detect polymorphism.Over 7 alleles(7-12) were found.Heterozygosity is below 70% in only one locus—CSF1PO. There are 9 complete chimeras of which one died from GVHD.Conclusion: Chimerism after transplantation can be detected successfully with the five tetranucleotide repeat sequences.Every kind of chimerism has its prognosis.As cells of donor-derived decrease, prognosis is worse.