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Expressional Analysis of Immunoglobulin D in Cattle (Bos taurus) a Large Domesticated Ungulate

机译:免疫球蛋白d的写意分析牛(普通牛)大型有蹄类动物驯养

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摘要

For decades, it has remained unknown whether artiodactyls, such as cattle, pigs, and sheep, express immunoglobulin D (IgD), although the δ gene was identified in these species nearly 10 years ago. By developing a mouse anti-bovine IgD heavy chain monoclonal antibody (13C2), we show that secreted bovine IgD was present mainly as a monomer in serum and was heavily glycosylated by N-linked saccharides. Nonetheless, IgD was detectable in some but not all of the Holstein cattle examined. Membrane-bound IgD was detected in the spleen by western blotting. Flow cytometric analysis demonstrated that IgD-positive B cells constituted a much lower percentage of B cells in the bovine spleen (∼6.8% of total B cells), jejunal Peyer's patches (∼0.8%), and peripheral blood leukocytes (∼1.2%) than in humans and mice. Furthermore, IgD-positive B cells were almost undetectable in bovine bone marrow and ileal Peyer's patches. We also demonstrated that the bovine δ gene can be expressed via class switch recombination. Accordingly, bovine δ germline transcription, which involves an Iδ exon and is highly homologous to Iμ, was confirmed. However, we could not identify an Iδ promoter, despite bovine Eμ demonstrating both enhancer and promoter activity. This study has answered a long-standing question in cattle B cell biology and significantly contributes to our understanding of B cell development in this species.
机译:几十年来,尽管诸如牛,猪和绵羊之类的关节细菌是否表达免疫球蛋白D(IgD),仍是一个未知数,尽管δ基因已在近十年前被发现。通过开发小鼠抗牛IgD重链单克隆抗体(13C2),我们显示分泌的牛IgD主要作为单体存在于血清中,并被N-连接的糖类严重糖基化。尽管如此,在部分但并非全部的荷斯坦牛中都可检测到IgD。通过蛋白质印迹在脾脏中检测到膜结合的IgD。流式细胞仪分析表明,IgD阳性B细胞在牛脾(占总B细胞的约6.8%),空肠Peyer斑块(约0.8%)和外周血白细胞(约1.2%)中所占的B细胞百分比要低得多比人类和老鼠此外,在牛骨髓和回肠Peyer斑中几乎检测不到IgD阳性B细胞。我们还证明了牛δ基因可以通过类开关重组表达。因此,证实了涉及Iδ外显子并且与Iμ高度同源的牛δ种系转录。然而,尽管牛Eμ表现出增强子和启动子活性,我们仍无法鉴定Iδ启动子。这项研究回答了牛B细胞生物学中一个长期存在的问题,并为我们对这种物种B细胞发育的理解做出了重要贡献。

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