Molecular and functional characterization of porcine Siglec-3/CD33 and analysis of its expression in blood and tissues
Molecular and functional characterization of porcine Siglec-3/CD33 and analysis of its expression in blood and tissues
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DOI：
10.1016/j.dci.2015.04.002
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发表时间：
2015-08-01
影响因子：
2.9
通讯作者：
Dominguez, J.
中科院分区：
文献类型：
--
作者：
Alvarez, B.;Escalona, Z.;Dominguez, J.
从猪小肠文库获得编码与人Siglec-3/CD 33具有同源性的380 α-α 1型跨膜蛋白的cDNA克隆。蛋白质序列的分析确定了两个免疫球蛋白样结构域，一个跨膜区，和一个羧基末端尾与两个酪氨酸为基础的信号基序。Siglec-3转染的CHO细胞与聚丙烯酰胺糖缀合物的结合测定显示对α 2-6-连接的唾液酸的偏好。使用针对含有两个Ig样结构域的片段产生的mAb，发现猪Siglec-3在单核细胞和粒细胞及其骨髓前体上表达。在淋巴结、脾和肺泡巨噬细胞中也检测到。在非还原和还原条件下，MAbs从粒细胞裂解物中免疫沉淀51-60 kDa的蛋白质。MAb还用于分析Siglec-3对骨髓和血细胞的功能活性。单克隆抗体与Siglec-3的结合对细胞增殖或细胞因子产生无明显影响。(C)2015爱思唯尔有限公司版权所有。
A cDNA clone encoding a 380 a-a type 1 transmembrane protein with homology to human Siglec-3/ CD33 was obtained from a swine small intestine library. An analysis of protein sequence identified two immunoglobulin-like domains, a transmembrane region, and a carboxi-terminal tail with two tyrosine-based signalling motifs. Binding assays of Siglec-3 transfected CHO cells to polyacrylamide glycoconjugates showed a preference for alpha 2-6-linked sialic acids. Using mAbs raised against a fragment containing the two Ig-like domains, porcine Siglec-3 was found to be expressed on monocytes and granulocytes, and their bone marrow precursors. It was also detected in lymph node, splenic and alveolar macrophages. MAbs immunoprecipitated, from granulocyte lysates, a protein of 51-60 kDa under both non-reducing and reducing conditions. MAbs were also used to analyse functional activity of Siglec-3 on bone marrow and blood cells. Engagement of Siglec-3 by mAb had no apparent effect on cell proliferation or cytokine production. (C) 2015 Elsevier Ltd. All rights reserved.