Detection of minimal disease in hematopoietic malignancies of the B-cell lineage by using third-complementarity-determining region (CDR-III)-specific probes.
Detection of minimal disease in hematopoietic malignancies of the B-cell lineage by using third-complementarity-determining region (CDR-III)-specific probes.
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使用第三互补决定区 (CDR-III) 特异性探针检测 B 细胞谱系造血系统恶性肿瘤中的微小疾病。
DOI：
10.1073/pnas.86.13.5123
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发表时间：
1989
影响因子：
11.1
通讯作者：
Rovera,G
中科院分区：
文献类型：
--
作者：
Yamada,M;Hudson,S;Tournay,O;Bittenbender,S;Shane,SS;Lange,B;Tsujimoto,Y;Caton,AJ;Rovera,G
大约80%的B细胞系血液系统恶性肿瘤只携带一到两个免疫球蛋白重链基因重排，表明它们是克隆性起源的。在B细胞定位过程中，由于重链基因片段的各种可变区、多样性和连接区的重组而导致的这些重排导致了一个称为互补决定区III(CDR-III)的区域。由于广泛的体细胞突变，这个区域包含了重链片段的多样性区域，为每个B细胞克隆提供了一个DNA编码的签名。用合适的引物和聚合酶链式反应从前B细胞急性淋巴细胞白血病的DNA中扩增出CDR-III序列。这些序列被用来产生诊断探针，这些探针只与白血病细胞的扩增CDR-III杂交，这些序列就是从这些细胞获得的。用这些探针与其他细胞稀释1：10000即可检测到白血病细胞。通过将扩增的CDR-III克隆到重组文库中，对1例急性淋巴细胞白血病反复复发和缓解的骨髓样本中残留的白血病细胞进行了准确的定量。在持续超过7个月的临床缓解期间，骨髓中恶性细胞的比例超过1/1000个细胞。这些发现表明，定制的诊断探针将有助于对临床缓解期急性淋巴细胞白血病患者的恶性细胞进行准确的定量，并将有助于研究残留白血病细胞数量少或多在疾病演变中的生物学意义。
Approximately 80% of hematopoietic malignancies of the B-cell lineage carry only one or two immunoglobulin heavy chain gene rearrangements indicating their clonal origin. These rearrangements due to the recombination of various variable, diversity, and joining regions of the heavy-chain gene segments during B-cell commitment result in a region called complementarity-determining region III (CDR-III). This region, which encompasses the diversity region of the heavy-chain segment, because of extensive somatic mutations, provides a DNA-encoded signature specific for each B-cell clone. CDR-III sequences were obtained from DNA of pre-B-cell acute lymphoblastic leukemia by using suitable primers and the polymerase chain reaction. The sequences were used to generate diagnostic probes that hybridized only to the amplified CDR-III of leukemic cells from which the sequences were derived. With these probes, leukemic cells could be detected when diluted 1:10,000 with other cells. By cloning the amplified CDR-III into recombinant libraries residual leukemic cells were accurately quantitated in bone-marrow samples from repeated relapses and remissions in one case of acute lymphoblastic leukemia. During a clinical remission lasting greater than 7 mo, malignant cells were present in marrow at greater than 1 per 1000 cells. These findings indicate that custom-made diagnostic probes will be useful in accurate quantitation of malignant cells in acute lymphoblastic leukemia patients in clinical remission and will allow investigation of the biological significance of low or high numbers of residual leukemic cells in evolution of that disease.