Moredun Research Institute, Pentlands Science Park, Bush Loan, Penicuik, Scotland, UK.
Parasite Immunol. 2010 Jul;32(7):503-11. doi: 10.1111/j.1365-3024.2010.01215.x.
A macrophage migration inhibitory factor (MIF)-like molecule, Tci-MIF-1, was isolated from Teladorsagia circumcincta and subjected to detailed characterization. A cDNA representing Tci-mif-1 was isolated following its identification in third-stage larvae (L3)-enriched cDNA population. Sequencing of the cDNA indicated a 348-bp open reading frame (ORF) with the closest orthologue being a MIF derived from the human hookworm Ancylostoma ceylanicum. Messenger RNA (mRNA) representing the Tci-MIF-1 transcript was detected in eggs, L3 and adult stages of T. circumcincta. The transcript was also present, but to a lesser extent in fourth-stage larvae (L4). Detection of Tci-MIF-1 protein in T. circumcincta developmental stages reflected the transcript levels identified by reverse transcriptase-PCR. Using immunohistochemistry, the Tci-MIF-1 protein was shown to have a diffuse distribution in L3 tissue, and in L4 and adult stages, the protein was localized to the nematode gut. A recombinant version of Tci-MIF-1 was produced, and enzymic assays indicated that this recombinant protein and a somatic extract of L3 possessed dopachrome tautomerase activity as has been observed previously in other MIF-like molecules. Neither native, purified Tci-MIF nor recombinant Tci-MIF-1 dramatically influenced the in vitro migration of sheep monocytes.
从捻转血矛线虫中分离出一种巨噬细胞移动抑制因子(MIF)样分子 Tci-MIF-1,并对其进行了详细的特征描述。在富含第三期幼虫(L3)的 cDNA 群体中鉴定出 Tci-mif-1 后,分离出了其 cDNA。cDNA 测序表明,该 cDNA 具有一个 348bp 的开放阅读框(ORF),其最接近的同源物是一种来源于人类钩虫Ancylostoma ceylanicum 的 MIF。在捻转血矛线虫的卵、L3 和成虫阶段均检测到代表 Tci-MIF-1 转录本的 mRNA。在 L4 期幼虫中也存在该转录本,但程度较低。逆转录 PCR 鉴定的转录本水平反映了 Tci-MIF-1 蛋白在捻转血矛线虫发育阶段的存在。通过免疫组织化学,Tci-MIF-1 蛋白在 L3 组织中呈弥散分布,在 L4 和成虫阶段,该蛋白定位于线虫肠道。产生了重组形式的 Tci-MIF-1,酶活性测定表明,这种重组蛋白和 L3 的体细胞提取物具有多巴色素互变异构酶活性,这在其他 MIF 样分子中已有观察到。天然的、纯化的 Tci-MIF 或重组 Tci-MIF-1 均未显著影响绵羊单核细胞的体外迁移。