Molecular cloning of a novel human CC chemokine (eotaxin-3) that is a functional ligand of CC chemokine receptor 3
Molecular cloning of a novel human CC chemokine (eotaxin-3) that is a functional ligand of CC chemokine receptor 3
复制标题
DOI：
10.1074/jbc.274.39.27975
复制
发表时间：
1999-09-24
影响因子：
4.8
通讯作者：
Yoshie, O
中科院分区：
文献类型：
--
作者：
Kitaura, M;Suzuki, N;Yoshie, O
此前，我们将新型 CC 趋化因子骨髓祖细胞抑制因子 2 (MPIF-2)/eotaxin-2 定位到染色体 7q11.23（Nomiyama, H.、Osborne, L. R.、Imai, T.、Kusuda, J、Miura, R.、Tsui, L.-C. 和 Yoshie, O. (1998) Genomics 49， 339-340）。由于趋化因子基因往往聚集在一起，未知的趋化因子可能存在于映射到新染色体位点的趋化因子附近。在这一假设的推动下，我们分析了包含 MPIF-2/eotaxin-2 (SCYA24) 基因的基因组区域，并鉴定了一种新的 CC 趋化因子，称为 eotaxin-3。 MPIF-2/eotaxin-2 (SCYA24) 和eotaxin-3 (SCYA26) 的基因定位在大约40 kilobase 的区域内。通过Northern blot 分析，eotaxin-3 mRNA 在心脏和卵巢中组成型表达。我们在杆状病毒表达系统中产生了重组eotaxin-3。 Eotaxin-3 特异性地在表达 CC 趋化因子受体 3 (CCR3) 的 L1.2 细胞中诱导瞬时钙动员，EC50 为 3 nM。 Eotaxin-3 竞争 I-125-eotaxin 与表达 CCR3 的 L1.2 细胞的结合，IC50 为 13 nM。 Eotaxin-3 在高浓度时对正常外周血嗜酸性粒细胞和嗜碱性粒细胞具有趋化作用。总的来说，eotaxin-3 是 CCR3 的另一个功能性配体。然而，eotaxin-3 作为 CCR3 配体的效力似乎比 eotaxin 低 10 倍。eotaxin-3 的鉴定将进一步促进我们对嗜酸性粒细胞运输控制和其他 CCR3 介导的生物现象的理解。本研究中使用的策略也可能适用于其他未知趋化因子基因的鉴定。
Previously, we mapped the novel CC chemokine myeloid progenitor inhibitory factor 2 (MPIF-2)/eotaxin-2 to chromosome 7q11.23 (Nomiyama, H., Osborne, L. R., Imai, T., Kusuda, J,, Miura, R., Tsui, L.-C., and Yoshie, O. (1998) Genomics 49, 339-340). Since chemokine genes tend to be clustered, unknown chemokines may be present in the vicinity of those mapped to new chromosomal loci. Prompted by this hypothesis, we analyzed the genomic region containing the gene for MPIF-2/eotaxin-2 (SCYA24) and have identified a novel CC chemokine termed eotaxin-3. The genes for MPIF-2/eotaxin-2 (SCYA24) and eotaxin-3 (SCYA26) are localized within a region of similar to 40 kilobases, By Northern blot analysis, eotaxin-3 mRNA was constitutively expressed in the heart and ovary. We have generated recombinant eotaxin-3 in a baculovirus expression system. Eotaxin-3 induced transient calcium mobilization specifically in CC chemokine receptor 3 (CCR3)-expressing L1.2 cells with an EC50 of 3 nM. Eotaxin-3 competed the binding of I-125-eotaxin to CCR3-expressing L1.2 cells with an IC50 of 13 nM. Eotaxin-3 was chemotactic for normal peripheral blood eosinophils and basophils at high concentrations. Collectively, eotaxin-3 is yet another functional ligand for CCR3. The potency of eotaxin-3 as a CCR3 ligand seems, however, to be similar to 10-fold less than that of eotaxin, Identification of eotaxin-3 will further promote our understanding of the control of eosinophil trafficking and other CCR3-mediated biological phenomena. The strategy used in this study may also be applicable to identification of other unknown chemokine genes.