Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
J Immunol. 2010 Nov 15;185(10):6085-95. doi: 10.4049/jimmunol.1001423. Epub 2010 Oct 18.
To assess the extent and nature of somatic categorical selection of CDR-3 of the Ig H chain (CDR-H3) content in peritoneal cavity (PerC) B cells, we analyzed the composition of V(H)7183DJCμ transcripts derived from sorted PerC B-1a, B-1b, and B-2 cells. We divided these sequences into those that contained N nucleotides (N(+)) and those that did not (N(-)) and then compared them with sequences cloned from sorted IgM(+)IgD(+) B cells from neonatal liver and both wild-type and TdT-deficient adult bone marrow. We found that the PerC B-1a N(-) repertoire is enriched for the signatures of CDR-H3 sequences present in neonatal liver and shares many features with the B-1b N(-) repertoire, whereas the PerC B-1a N(+), B-1b N(+), and B-2 N(+) repertoires are enriched for adult bone marrow sequence signatures. However, we also found several sequence signatures that were not shared with other mature perinatal or adult B cell subsets but were either unique or variably shared between the two or even among all three of the PerC subsets that we examined. These signatures included more sequences lacking N nucleotides in the B-2 population and an increased use of D(H) reading frame 2, which created CDR-H3s of greater average hydrophobicity. These findings provide support for both ontogenetic origin and shared Ag receptor-influenced selection as the mechanisms that shape the unique composition of the B-1a, B-1b, and B-2 repertoires. The PerC may thus serve as a general reservoir for B cells with Ag binding specificities that are uncommon in other mature compartments.
为了评估 Ig H 链(CDR-H3)的 CDR-3 内容在腹腔(PerC)B 细胞中的体细胞分类选择的程度和性质,我们分析了从分选的 B-1a、B-1b 和 B-2 细胞中衍生的 V(H)7183DJCμ 转录本的组成。我们将这些序列分为包含 N 核苷酸(N(+))和不包含 N 核苷酸(N(-))的序列,然后将它们与从新生儿肝脏分选的 IgM(+)IgD(+)B 细胞以及野生型和 TdT 缺陷型成年骨髓中克隆的序列进行比较。我们发现 PerC B-1a N(-) repertoire 富含存在于新生儿肝脏中的 CDR-H3 序列特征,并与 B-1b N(-) repertoire 具有许多共同特征,而 PerC B-1a N(+)、B-1b N(+)和 B-2 N(+) repertoire 则富含成人骨髓序列特征。然而,我们还发现了一些与其他成熟围产期或成年 B 细胞亚群不同的序列特征,但它们要么是独特的,要么在两个甚至所有三个我们检查的 PerC 亚群之间存在可变共享。这些特征包括 B-2 群体中缺乏 N 核苷酸的更多序列和 D(H)阅读框 2 的使用增加,这会产生平均疏水性更高的 CDR-H3。这些发现为胚胎起源和共同的 Ag 受体影响选择作为塑造 B-1a、B-1b 和 B-2 repertoire 独特组成的机制提供了支持。因此,PerC 可能是具有在其他成熟隔室中罕见的 Ag 结合特异性的 B 细胞的一般储库。