Molecular cloning of a novel human CC chemokine secondary lymphoid-tissue chemokine that is a potent chemoattractant for lymphocytes and mapped to chromosome 9p13
Molecular cloning of a novel human CC chemokine secondary lymphoid-tissue chemokine that is a potent chemoattractant for lymphocytes and mapped to chromosome 9p13
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DOI：
10.1074/jbc.272.31.19518
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发表时间：
1997-08-01
影响因子：
4.8
通讯作者：
Yoshie, O
中科院分区：
文献类型：
--
作者：
Nagira, M;Imai, T;Yoshie, O
通过搜索表达序列标签（EST）数据库，我们确定了部分cDNA序列可能编码一种新的人类CC趋化因子。我们确定了整个cDNA序列，它编码一个高度碱性的多肽，共134个氨基酸的23个氨基酸的推定信号肽。预测的111个氨基酸的成熟蛋白具有四个典型的半胱氨酸残基，并显示与其他人CC趋化因子的21-33%的同一性，但具有独特的约30个氨基酸的羧基末端延伸，其含有两个额外的半胱氨酸残基。该mRNA在淋巴结、阑尾和脾脏等组织中强烈表达。重组蛋白，这是由杆状病毒系统和纯化的同质性，是一种高效的化学引诱剂，某些人的T细胞系和一个非常有效的新鲜分离的外周血淋巴细胞和培养的正常T细胞的植物血凝素和白细胞介素2扩大。然而，与大多数其他CC趋化因子不同，这种新型趋化因子对单核细胞或中性粒细胞没有趋化性，表明它对淋巴细胞具有特异性。根据这些结果，我们将这种新的CC趋化因子命名为次级淋巴组织趋化因子SLC。SLC与分泌形式的碱性磷酸酶（SLC-SEAP）融合用于表征SLC受体。SLC-SEAP与新鲜分离的淋巴细胞的结合被SLC阻断（IC 50，0.12 nM），但迄今为止未被任何其他测试的CC趋化因子阻断，这表明静息淋巴细胞表达一类对SLC高度特异性的受体。通过使用体细胞杂交，辐射杂交，和选择的酵母和细菌人工染色体克隆，我们映射SLC基因（SCYA 21）在染色体9 p13和染色体标记，D9 S1978（WI-8765）和AFM 326 vd 1之间，其中另一种新的CC趋化因子ELC从EBI 1配体趋化因子（SCYA 19）的基因也存在。总的来说，SLC是一种新的CC特异性淋巴细胞趋化因子，并与ELC一起，构成了一组新的定位于染色体9 p13的趋化因子。
By searching the Expressed Sequence Tag (EST) data base, we identified partial cDNA sequences potentially encoding a novel human CC chemokine. We determined the entire cDNA sequence which encodes a highly basic polypeptide of 134 amino acids total with a putative signal peptide of 23 amino acids. The predicted mature protein of 111 amino acids has the four canonical cysteine residues and shows 21-33% identity to other human CC chemokines, but has a unique carboxyl-terminal extension of about 30 amino acids which contains two extra cysteine residues. The mRNA was expressed strongly in tissues such as the lymph nodes, appendix, and spleen. The recombinant protein, which was produced by the baculovirus system and purified to homogeneity, was a highly efficient chemoattractant for certain human T cell lines and a highly potent one for freshly isolated peripheral blood lymphocytes and cultured normal T cells expanded by phytohemagglutinin and interleukin 2. Unlike most other CC chemokines, however, this novel chemokine was not chemotactic for monocytes or neutrophils, suggesting that it is specific for lymphocytes. From these results, we designated this novel CC chemokine as SLC from secondary lymphoid-tissue chemokine. SLC fused with the secreted form of alkaline phosphatase (SLC-SEAP) was used to characterize the SLC receptor. Binding of SLC-SEAP to freshly isolated lymphocytes was blocked by SLC (IC50, 0.12 nM) but not by any other CC chemokine so far tested, suggesting that resting lymphocytes express a class of receptors highly specific for SLC. By using somatic cell hybrids, radiation hybrids, and selected yeast and bacterial artificial chromosome clones, we mapped the SLC gene (SCYA21) at chromosome 9p13 and between chromosomal markers, D9S1978(WI-8765) and AFM326vd1, where the gene for another novel CC chemokine termed ELC from EBI1-ligand chemokine (SCYA19) also exists. Collectively, SLC is a novel CC chemokine specific for lymphocytes and, together with ELC, constitutes a new group of chemokines localized at chromosome 9p13.