Detection of minimal residual disease in acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor delta chain sequences.
Detection of minimal residual disease in acute lymphoblastic leukemia by in vitro amplification of rearranged T-cell receptor delta chain sequences.
复制标题
通过体外扩增重排的 T 细胞受体 δ 链序列来检测急性淋巴细胞白血病中的微小残留病。
作者：
Thomas E. Hansen;Shouhei Yokota;Claus R. Bartram
人类T细胞受体（TCR）δ链的多样性主要来源于高度的连接变异性，因为只有有限数量的种系元件是可用的。由于使用两个D δ元件和N核苷酸的广泛掺入，在V.J连接处的这种非凡的多样性构成了表现出重排的TCR δ基因的细胞群体的特异性克隆标记。为此，我们在体外扩增聚合酶链反应（PCR）的TCR δ连接区的5个急性淋巴细胞白血病（ALL），分离各自的DNA片段，并直接使用它们作为克隆特异性探针。PCR技术和与克隆特异性探针杂交的组合允许在所有五个病例中以1：100，000稀释检测白血病DNA。此外，我们能够研究一个ALL患者11个月后实现连续完全缓解。传统的Southern印迹分析未能检测到重排的TCR基因在这个阶段。然而，残留的白血病细胞可以很容易地检测到PCR技术。我们的结论是，这里提出的策略是一个非常敏感的工具，以检测微小残留病在一个显着比例的人类淋巴瘤。
Human T-cell receptor (TCR) delta-chain diversity mainly originates from high junctional variability, since only a limited number of germline elements is available. This extraordinary diversity at the V.J junction, due to the use of two D delta elements and extensive incorporation of N nucleotides, constitutes a specific clonal marker for cell populations exhibiting rearranged TCR delta genes. To this end we amplified in vitro by polymerase chain reaction (PCR) the TCR delta junctional region of five acute lymphoblastic leukemias (ALL), isolated respective DNA fragments, and used them directly as clonospecific probes. The combination of PCR technology and hybridization to clonospecific probes permitted the detection of leukemia DNA at dilution of 1:100,000 in all five cases. Moreover, we were able to investigate one of the ALL patients 11 months after achieving continuous complete remission. Conventional Southern blot analysis failed to detect rearranged TCR genes at this stage. However, residual leukemic cells could readily be detected by PCR technique. We conclude that the strategy proposed here is a very sensitive tool to detect minimal residual disease in a significant proportion of human lymphoid neoplasias.
通过转移到电泳室中的 DEAE 纸上从凝胶中回收 DNA 片段。
影响因子：
2.9
作者：
Danner,DB
通讯作者：
Danner,DB
重排的人类 T 细胞受体 delta 基因的广泛连接多样性。
DOI：
10.1126/science.3259726
发表时间：
1988
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Hata,S;Satyanarayana,K;Devlin,P;Band,H;McLean,J;Strominger,JL;Brenner,MB;Krangel,MS
通讯作者：
Krangel,MS