Induced rearrangement of kappa genes in the BLIN-1 human pre-B cell line correlates with germline J-C kappa and V kappa transcription.
Induced rearrangement of kappa genes in the BLIN-1 human pre-B cell line correlates with germline J-C kappa and V kappa transcription.
复制标题
BLIN-1 人类前 B 细胞系中 kappa 基因的诱导重排与种系 J-C kappa 和 V kappa 转录相关。
DOI：
10.1084/jem.173.3.639
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
VanNess,B
中科院分区：
文献类型：
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作者：
Martin,D;Huang,RQ;LeBien,T;VanNess,B
人类Pre-B急性淋巴细胞白血病细胞系Blin-1在体外发生了kappa轻链重排，是分析Pre-B到B细胞分化的有价值的资源。我们通过对几个独立衍生的表达kappa的亚克隆进行DNA重排和V kappa基因的使用来研究Blin-1的重组潜力。对5个表达kappa的亚克隆(均具有相同重链重排)的DNA杂交分析表明，kappa轻链重排事件是相互独立的。用识别四个不同V kappa家族的探针进行Northern印迹分析，发现两个亚克隆使用最接近的V kappa(V Kappa IV)，一个亚克隆使用V kappa I，一个亚克隆使用V kappa II。通过聚合酶链式反应分析，我们检测到重排的V-J-C kappa基因的转录本，以及在诱导Blin-1细胞重排Kappa基因的胚系J-C kappa和V kappa的转录本。Kappa胚系转录本也在胎儿肝脏和骨髓中正常发育的B细胞群中检测到。我们的集体结果表明：(A)Blin-1可被诱导重排kappa基因，这与胚系kappa基因转录本的表达有关，该转录本可能在激活或靶向基因重排中发挥作用；以及(B)代表不同kappa家族的V基因的主动重排和使用表明，与小鼠一样，人类的曲谱系多样化发生在固定的重链重排存在的情况下。
The human pre-B acute lymphoblastic leukemia cell line, BLIN-1, has been previously shown to undergo kappa light chain rearrangement in vitro, making it a valuable resource for analyzing pre-B to B cell differentiation. We have examined the recombination potential of BLIN-1 by characterizing several independently derived kappa-expressing subclones for DNA rearrangement and V kappa gene usage. Analysis of five kappa-expressing subclones (all having the same heavy chain rearrangement) demonstrated independent kappa light chain rearrangement events by DNA hybridization analysis. Northern blot analysis using probes recognizing the four different V kappa families revealed that two subclones used the most proximal V kappa (V kappa IV), one subclone used a V kappa I, and one subclone used a V kappa II. By polymerase chain reaction analyses, we detected transcripts from rearranged V-J-C kappa genes as well as transcripts from germline J-C kappa and V kappa in BLIN-1 cells induced to rearrange the kappa locus. kappa germline transcripts were also detected in normal developing B cell populations in fetal liver and bone marrow. Our collective results indicate that: (a) BLIN-1 can be induced to rearrange the kappa locus, and this correlates with the expression of germline kappa locus transcripts that may play a role in activating or targeting gene rearrangement; and (b) active rearrangement and usage of V genes representing different kappa families suggest that, like in the mouse, repertoire diversification in humans occurs in the presence of a fixed heavy chain rearrangement.