Prado Carmen, Rodríguez Mercedes, Cortegano Isabel, Ruiz Carolina, Alía Mario, de Andrés Belén, Gaspar María Luisa
Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain.
J Innate Immun. 2014;6(4):499-514. doi: 10.1159/000358237. Epub 2014 Mar 6.
The diversity in antibody repertoire relies on different B cell populations working efficiently to fulfil distinct specific functions. We recently described an innate-like CD19(+)CD45R(-/lo) (19(+)45R(lo)) cell population in postnatal unstimulated adult mice, a heterogeneous population containing cells expressing immunoglobulin M (IgM) and others behaving as differentiated mature B lymphocytes (intracytoplasmic IgG1, AID(+), Blimp-1(+)RAG2(-)). In the present study, we characterized the Ig repertoire expressed by splenic 19(+)45R(lo) cells, assuming that they would bear a restricted repertoire biased for germline rearrangements and low mutation rates similar to other innate-like cells. Sequences from 19(+)45R(lo) cells displayed a variety of V, D and J regions, and the analysis of the CDR-H3 region revealed an intermediate overall CDR-H3 length and moderate hydrophobicity. Both IgM and switched sequences of PD15 19(+)45R(lo) cells had shorter CDR-H3 region and fewer non-template N nucleotides than adult sequences, as expected for profiles that correspond to an immature phenotype. Regarding the mutation rate in the VH regions, IgG1 sequences already carried a high rate of replacement mutations at PD15, which increased further in the sequences obtained from adult mice. Moreover, statistical models suggest that a proportion of the switched sequences in adult 19(+)45R(lo) cells had experienced antigen selection, unlike other innate-like B cell compartments.
抗体库的多样性依赖于不同的B细胞群体高效工作以履行不同的特定功能。我们最近在出生后未受刺激的成年小鼠中描述了一种天然样CD19(+)CD45R(-/lo)(19(+)45R(lo))细胞群体,这是一个异质群体,包含表达免疫球蛋白M(IgM)的细胞以及其他表现为分化成熟B淋巴细胞的细胞(胞质内IgG1、AID(+)、Blimp-1(+)RAG2(-))。在本研究中,我们对脾脏19(+)45R(lo)细胞表达的Ig库进行了表征,假设它们具有偏向种系重排和低突变率的受限库,类似于其他天然样细胞。来自19(+)45R(lo)细胞的序列显示出多种V、D和J区域,对CDR-H3区域的分析揭示了CDR-H3的整体长度处于中间水平且疏水性适中。PD15 19(+)45R(lo)细胞的IgM和转换序列的CDR-H3区域都比成年序列短,非模板N核苷酸也更少,这与对应不成熟表型的特征预期相符。关于VH区域的突变率,IgG1序列在PD15时就已经具有较高的替换突变率,在从成年小鼠获得的序列中进一步增加。此外,统计模型表明,成年19(+)45R(lo)细胞中的一部分转换序列经历了抗原选择,这与其他天然样B细胞区室不同。