Spiess S, Kuhröber A, Schirmbeck R, Reimann J
Institute of Microbiology, University of Ulm, FRG.
Eur J Immunol. 1992 Jul;22(7):1939-42. doi: 10.1002/eji.1830220740.
From bone marrow cells (BMC) of athymic nude mice, T cell receptor (TcR) alpha chain transcripts were selectively amplified by polymerase chain reaction (PCR) using V delta 2-, V delta 3-, V delta 4-, V delta 5-, V delta 6- and C alpha-specific primers. Amplified DNA fragments were cloned, and 32 randomly selected clones from 5 PCR were sequenced. Twenty-three distinct rearrangement events were detected, of which 87% (20/23) were in-frame. All five tested V delta genes (V delta 2, 3, 4, 5, 6) rearranged in-frame to J alpha-C alpha. N-region diversity in V delta-J alpha junctions present in most clones was limited to two to five nucleotides. P-nucleotide additions in this region were also detected. The V delta 5 gene located 3' of C delta in reversed transcriptional orientation was rearranged to J alpha by inversion. The J alpha usage pattern of the sequenced clones was strongly biased towards rearrangement of the most 5' genes (located nearest to C delta) of the J alpha cluster: the most 5' J alpha (J alpha TA1) was used by 30% of all clones, and 78% of all J alpha rearranged to V delta were located in the 5' 12 kb of the 60-kb J alpha cluster. As distinct V delta/C delta and V alpha/C alpha TcR usage patterns are prevalent in peripheral T cell populations, our data suggest that these TcR usage patterns results from repertoire selections operating in alpha beta and gamma delta T cell lineages, but not from preferential V delta-C delta and V alpha-C alpha rearrangement patterns.
利用Vδ2、Vδ3、Vδ4、Vδ5、Vδ6和Cα特异性引物,通过聚合酶链反应(PCR)从无胸腺裸鼠的骨髓细胞(BMC)中选择性扩增T细胞受体(TcR)α链转录本。扩增的DNA片段被克隆,并对来自5次PCR的32个随机选择的克隆进行测序。检测到23个不同的重排事件,其中87%(20/23)为框内重排。所有五个测试的Vδ基因(Vδ2、3、4、5、6)均框内重排至Jα-Cα。大多数克隆中Vδ-Jα连接处的N区多样性限于2至5个核苷酸。该区域也检测到P核苷酸添加。转录方向相反位于Cδ 3'端的Vδ5基因通过倒位重排至Jα。测序克隆的Jα使用模式强烈偏向于Jα簇最5'端基因(最靠近Cδ)的重排:所有克隆中有30%使用最5'端的Jα(Jα TA1),所有重排至Vδ的Jα中有78%位于60 kb Jα簇的5'端12 kb内。由于在外周T细胞群体中不同的Vδ/Cδ和Vα/Cα TcR使用模式普遍存在,我们的数据表明这些TcR使用模式是由αβ和γδ T细胞谱系中的库选择导致的,而非来自优先的Vδ-Cδ和Vα-Cα重排模式。