State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China.
PLoS One. 2013 Jul 22;8(7):e69284. doi: 10.1371/journal.pone.0069284. Print 2013.
Insect gut immunity is the first line of defense against oral infection. Although a few immune-related molecules in insect intestine has been identified by genomics or proteomics approach with comparison to well-studied tissues, such as hemolymph or fat body, our knowledge about the molecular mechanism underlying the gut immunity which would involve a variety of unidentified molecules is still limited. To uncover additional molecules that might take part in pathogen recognition, signal transduction or immune regulation in insect intestine, a T7 phage display cDNA library of the silkworm midgut is constructed. By use of different ligands for biopanning, Translationally Controlled Tumor Protein (TCTP) has been selected. BmTCTP is produced in intestinal epithelial cells and released into the gut lumen. The protein level of BmTCTP increases at the early time points during oral microbial infection and declines afterwards. In vitro binding assay confirms its activity as a multi-ligand binding molecule and it can further function as an opsonin that promotes the phagocytosis of microorganisms. Moreover, it can induce the production of anti-microbial peptide via a signaling pathway in which ERK is required and a dynamic tyrosine phosphorylation of certain cytoplasmic membrane protein. Taken together, our results characterize BmTCTP as a dual-functional protein involved in both the cellular and the humoral immune response of the silkworm, Bombyx mori.
昆虫肠道免疫是抵御口腔感染的第一道防线。虽然通过基因组学或蛋白质组学方法与研究较为充分的组织(如血淋巴或脂肪体)进行比较,已经鉴定出几种与昆虫肠道相关的免疫相关分子,但我们对涉及各种未鉴定分子的肠道免疫的分子机制的了解仍然有限。为了揭示可能参与昆虫肠道中病原体识别、信号转导或免疫调节的其他分子,构建了家蚕中肠的 T7 噬菌体展示 cDNA 文库。通过使用不同的配体进行生物淘选,已选择出翻译控制肿瘤蛋白(TCTP)。BmTCTP 在肠上皮细胞中产生并释放到肠腔中。在口腔微生物感染的早期时间点,BmTCTP 的蛋白水平增加,随后下降。体外结合实验证实其作为多配体结合分子的活性,并且可以进一步作为促进微生物吞噬作用的调理素。此外,它可以通过需要 ERK 的信号通路诱导抗菌肽的产生,并使某些细胞质膜蛋白发生动态酪氨酸磷酸化。总之,我们的研究结果表明 BmTCTP 是一种双功能蛋白,参与家蚕(Bombyx mori)的细胞和体液免疫反应。