Victor K D, Vu K, Feeney A J
Scripps Research Institute, Department of Immunology, La Jolla, CA 92037.
J Immunol. 1994 Apr 1;152(7):3467-75.
Among all adult T and B cell Ag receptor chains, only Ig light chains lack N regions. It is thought that this is due to the fact that light chain genes rearrange after heavy chain genes, and that terminal deoxynucleotidyl transferase, the enzyme that adds N regions, is not longer expressed at that stage. However, this concept has been challenged recently by the demonstration that 3 to 10% of B cell precursors (CD43+B220+) appear to rearrange their light chains at approximately the same time as they undergo VH-->DJ rearrangements. To examine N region addition in B cell precursors undergoing early kappa-chain rearrangement, we PCR amplified rearranged V kappa 21 genes from the CD43+B220+ bone marrow cells and compared them to sequences obtained from whole bone marrow and spleen. Unexpectedly, all three populations showed approximately 10% N region containing junctions, most consisting of only one N nucleotide. Thus, even the B cell precursors that rearrange light chains at this early stage of development lack much N region diversity. Twelve percent of the sequences unambiguously contained P regions, which were from 1 to 5 nucleotides in length. All but 2 of the 41 productive rearrangements had the commonly observed CDR3 length of nine amino acids. Many (71%) of the sequences were out of frame. CDR3 length was very restricted in nonproductive rearrangements too, and deletion of nucleotides from V kappa and J kappa gene segments was limited. Thus, even at the level of nonproductive rearrangements, junctional diversity is minimal for kappa-chains.
在所有成年T细胞和B细胞抗原受体链中,只有免疫球蛋白轻链缺乏N区。据认为,这是因为轻链基因在重链基因之后发生重排,并且添加N区的末端脱氧核苷酸转移酶在该阶段不再表达。然而,最近这一概念受到了挑战,因为有证据表明3%至10%的B细胞前体(CD43+B220+)在进行VH→DJ重排时似乎大约同时重排其轻链。为了研究早期κ链重排的B细胞前体中N区的添加情况,我们通过聚合酶链反应(PCR)从CD43+B220+骨髓细胞中扩增重排的Vκ21基因,并将它们与从全骨髓和脾脏获得的序列进行比较。出乎意料的是,所有这三个群体都显示出约10%的含有N区的连接,大多数仅由一个N核苷酸组成。因此,即使在发育的这个早期阶段重排轻链的B细胞前体也缺乏太多的N区多样性。12%的序列明确含有P区,其长度为1至5个核苷酸。41个有效重排中除2个外,所有重排的互补决定区3(CDR3)长度均为常见的9个氨基酸。许多(71%)序列是移码的。在无效重排中,CDR3长度也受到非常严格的限制,并且Vκ和Jκ基因片段的核苷酸缺失是有限的。因此,即使在无效重排水平,κ链的连接多样性也是最小的。